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 BACTERIA OF THE sputa: 
 
 MlYPTaaAMIC FLORA 
 
 THBiMQUTH 
 

 ■^ 
 
 CORNELL UNIVERSITY. 
 
 V ^5J^- THE 
 
 THE GIFT OF 
 
 RO SWELL P. FLOWER 
 
 FOR THE USE OF 
 
 THEN. Y. STATE VETERINARY COLLEGE. 
 
 1897 
 
The original of tiiis book is in 
 tine Cornell University Library. 
 
 There are no known copyright restrictions in 
 the United States on the use of the text. 
 
 http://www.archive.org/cletails/cu31924000223481 
 
BACTERIA OF THE SPUTA 
 
 AND 
 
 CRYPTOGAMIC FLORA OF THE MOUTH. 
 
LIBRARY. 
 
 BACTERIA OF THE SPUTA 
 
 AND 
 
 GRYPTOGAMIC FLORA OF THE MOUTH. 
 
 BY 
 
 FILANDRO yiCENTINI, M.D. 
 
 TRANSLATED BY 
 
 Rev. E. J. STUTTER and Professor E. SAIEGHI, 
 
 FROM THE 
 
 "Aui della R. Accademia Medico- Chirurgica" of Naples. 
 
 Xonfton : 
 
 BAILLIERE, TINDALL, & COX, 20-21 KING WILLIAM ST., STRAND. 
 
 1897. 
 
ERRATA. 
 — o — 
 
 Page 4, line II, for "these" read "true." 
 
 Page 5, line i, for "pus, corpuscles'' read "pus corpuscles.'' 
 
 Page 8, line 3 from bottom, yor "micrococci" read " pneumococci." 
 
 Page 10, line 4, for "per hour " read "in twenty-four hours." 
 
 Page 10, last line, omit "p. 1882." 
 
 Page II, line 23, for "composition" read "decomposition."' 
 
 Page 14, line 8, for "k and k" read "k and k>." 
 
 Page 20, line 18, /or "a mixture of nitric acid and alcohol (l : 3)," read 
 "a mixture of nitric acid and water (i : 3), and, soon 
 after, absolute alcohol." 
 
 Pa^e 21, line 2, for "in ^" reati "in ^. '' 
 
 Page 66, line 12, for "these" read "there.'' 
 
 Page 95, line 27, for "hand" read "handle." 
 
 Page 109, line 35, for " which, although old, is not ours," read " which is 
 not ours, but old." 
 
 Page 117, line 3, for "x and x" read "x and x'." 
 
 Page 132, line i, /or "saccharine" read "saccharose." 
 

 Preface. 
 
 T will be readily admitted by all that great interest centres 
 about the study of the micro-organisms of the human 
 mouth. 
 
 To the botanist, bacteriologist, and pathologist, no less than to 
 the dental surgeon, the bacteria of the oral cavity offer a fertile 
 field of investigation and present many questions, the solution of 
 which has thus far been attempted in vain. 
 
 Up to the present, our investigations have been most success- 
 ful in the direction of the pathological action of the mouth-bac- 
 teria, while, on account of the great variety of micro-organisms 
 found in the mouth, and on account of the fact that many of them 
 have resisted all attempts at cultivation on artificial media, no 
 satisfactory classification of them has as yet been made or even 
 attempted. 
 
 Consequently, any communications or investigations which may 
 increase our knowledge of the flora of the human mouth, and 
 help to bring about a classification which may serve as a basis for 
 future work, or even any communication which serves to excite a 
 discussion of the question, is deserving of thorough attention on 
 the part of dentists, physicians, and natural scientists. 
 
 In this sense I recommend the Memoirs of Dr. Vicentini for 
 a careful study. It is not the place here to enter into a critical 
 examination of the views of the author ; I refer the reader, in this 
 conhection, to the paper by J. Howard Mummery, of London, 
 published in the Transactions of the Odoniological Society of Great 
 Britain, Vol. XXVI., No. 3, JanuarJ?, 1894. Suffice it to say that 
 the deductions of Dr. Vicentini, if confirmed, would bring about 
 
Vlli. rREFACK. 
 
 a complete revolution of the views at present adopted by bacterio- 
 logists, resulting from the immense amount of work which has 
 been done in this field for the last ten or fifteen years. 
 
 In order that my own position' may not be misunderstood, I 
 should add that, while attaching proper importance to the valuable 
 work done by Dr. Vicentini, I am somewhat sceptical as to the 
 correctness of his deductions. In the summing up of the author's 
 work, which he has, placed at my disposal in the English and 
 French languages, I have found nothing which in my opinion jus- 
 tifies the conclusion that not only all bacteria found in the human 
 mouth, but that even the pneumococcus, bacillus tuberculosis, 
 and gonococcus are simply derivations of the same normal organ- 
 ism. If we go so far, we may with equal show of reason include 
 the bacilli of diphtheria, typhus, cholera, etc., in fact, derive all 
 known species of bacteria from one common parent plant. 
 
 In regard to the actual microscopical observations of Dr. 
 Vicentini, Mummery (I.e.) reports having in one case found 
 appearances very similar to those which Dr. Vicentini compares to 
 grape-bunches. Personally, I have had no great difficulty in find- 
 ing masses of granular bodies (micrococci) sometimes apparently 
 accumulated about one central thread, sometimes about a mass of 
 threads. Very often these masses are shapeless ; in other cases 
 they are shaped like a club or boomerang (Mummery), or bear a 
 certain similarity to a bunch of grapes. As yet, I have not been 
 able to devote much time to the study of these bodies, and can 
 therefore offer no explanation whatever as to their signification. 
 
 In consideration of the importance of the question at issue, it 
 is to be hoped that the work of Dr. Vicentini will receive a careful 
 perusal, and that the investigations incited by it may bring about 
 a better understanding of the biology of the micro-organisms of 
 the human mouth than we have as yet been able to obtain. 
 
 W. D. Miller. 
 Berlin, 1896. 
 
CONTENTS. 
 
 — f>C* — 
 
 PAQK 
 
 Preface, by Prof. Miller vii. — viii. 
 
 FIRST MEMOIR. Translated by Rev. E. J. Stutter. 
 
 On the Sputa of Whooping Cough - 1 
 
 I. — Previous Researches on the Sputa of Whooping Cough 2 
 
 II. — A Short Account of Cases in which the Sputa have 
 
 been examined 9 
 
 III.— Notes on Myelin 22 
 
 IV. — Observations and Considerations on the Bacteria - 37 
 
 V. — Observations and Considerations on the Fungi 62 
 
 VI. — Hints on the Preparation and Staining of Specimens 68 
 
 Resume 71 
 
 SECOND MEMOIR. Translated by E. Saieghi. 
 
 Recent Bacteriological Researches on the Sputa : The Morpho- '' 
 
 logy and Biology of the Microbes of the Mouth 75 
 
 I. — Further Remarks on the Bacteria and Bacilli found in 
 
 the Sputa and in the Contents of the Mouth 76 
 
 II. — Summary of the Present Investigations and Methods 
 
 of Working 86 
 
 III.— Opinions hitherto held respecting the Organisms of 
 
 the Mouth, according to Miller 96 
 
 IV. — Remarks on the Morphology and Biology of Leptothrix 
 
 buccalis 115 
 
 Recapitulation 137 
 
 Bibliography 142 
 
 THIRD MEMOIR. Translated by E. Saieghi. 
 On Leptothrix roMemosa 145 
 
 I. — Introductory and Biographical Notices — Reference to 
 
 the previous Memoir - 145 
 
CONTENTS. 
 
 '^otKsr 
 
 PAGE 
 
 II.— New Researches on the Fructification of the Normal 
 
 Parasite of the Mouth (Leptothrix racernosa) - 16° 
 
 Conclusion ^°^ 
 
 Appendix, translated by E. Saieghi " 1°^ 
 
 Reason of writing this Appendix ■ " '-''wJ- 
 
 Doubts respecting the Bacterial Hypothesis 193 
 
 The Experiments of Pettenkofer on Cholera 199 
 
 Hypothesis of the Author on the Action of Bacteria 203 
 
 Further Researches on the Morphology of Leptothrix 217 
 
 Explanation of Plates - 225 
 
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Bacteria of tbe Sputa anb Cri^ptogamic 
 jflora of tbe fiDoutb. 
 
 FIRST MEMOIR. 
 ON THE SPUTA OF WHOOPING-COUGH. 
 
 Plate I. 
 
 On the presence of Myelin in the Sputa of Whooping-cough, and 
 on the Bacteria and Microphytes at times found therein, with 
 remarks on the Bacteria of Sputa in general, and hints as to 
 the Staining and Preparing of Specimens for the microscope. 
 
 Summary : 
 
 § I. — Previous researches on the sputa of Whooping-cough 
 (Cerasi, Poulet, Jansen, Ransome, Letzerich, Henke, 
 Tschamer, Biirger, Moncorvo, Barlow, Michael, 
 Afanasieff). 
 
 § 2. — Notes on the Clinical cases, in which the sputa have 
 been examined. Fikst case {Sputa of May lo, 
 May 12, June 5/ sputum in Pneumonia, July 31). 
 Second case ; Third ; Fourth ; Fifth , Sixth ; 
 Seventh ; Eighth ; Ninth ; and other Cases. 
 
SPUTA OF WHOOPING-COUGH. 
 
 § 3. — Myelin. General Remarks. Myelin in the sputa 
 IN Whooping-cough. In other sputa (i.e.. Normal 
 sputa in acute and chronic Bronchitis, in Pneumonia, 
 Pleurisy, Phthisis, and Hcemoptysis). Importance of 
 Myelin in Whooping-cough. Researches on the 
 Breath. 
 
 § 4. — Bacteria. Bacteria and BacilU found in Whooping- 
 cough. Bacteria and BaciUi in normal sputa, in nasal 
 mucus, in Bronchitis, in Pneumonia, in Pleurisy, in 
 Phthisis. 
 
 § 5 . — Fungi (Filaments found in Whooping-cough and their 
 fructification). 
 
 § 6. — Preparation and staining specimens (Gentian violet, 
 Solution of iodine. Picric acid. Methyl violet, Fuchsin, 
 Blue aniline. Carmine). 
 Conclusion. 
 
 § I. 
 
 PREVIOUS RESEARCHES ON THE SPUTA OF 
 
 WHOOPING-COUGH. 
 
 T'HE epidemic of Whooping-cough which broke out in the town 
 of Chieti, from April till July of the year 1888, afforded me 
 an opportunity of making some few researches on the 
 nature of the sputa in Whooping-cough. Feeling that the results 
 of such investigations may shed some light on questions regarding 
 the pathogenesis, phases, and peculiarities of disease, in so many 
 respects still obscure, I venture to offer a short account of them 
 in this memoir. 
 
 It will be well, first of all, to point out briefly what has been 
 done in this matter since 1867 up to the present time. 
 
 The first account of the microscopical examination of Sputa 
 in Whooping-cough appears in the monograph of Hagenbach, 
 given in Gerhardt's treatise on th^ Diseases of Children and in the 
 papers by Barlow* on the Nature of Whooping-cough, which 
 appeared in four numbers of the Lancet in 1886. 
 
 * W. H. Barlow On the Nature of Whooping-cough, Lancet, 1886, Vol. I., 
 pp. 870, 915, 966, and 1241. 
 
SPUTA OF WHOOPING-COUGH. 3 
 
 111 both these works mention is made of the researches of 
 Poulet, in 1867, who, whilst examining the breath of infants, found 
 a form of Monas ternio or Bacterium termo, as well as another of 
 Monas punctum, Bodo punctum, or Bacterium bacillus, which he 
 thought to be the origin of the disease In the second and fourth 
 sections I shall refer again to this.* 
 
 In the year 1868, Jansen, in his investigations, found bacteria 
 in the sputum of Whooping-cough, but they were not identical 
 with those described by Poulet, and he remained in doubt whether 
 they were specific indications of Whooping-cough or simply 
 microbes of the mouth. + 
 
 In the year 1870, Ransome % brought before the Manchester 
 Literary and Philosophical Society the first notice of the presence 
 of a Conferva similar to Penicillium glaucum in the breath of 
 subjects in Whooping-cough. His researches were corroborated, 
 as regards measles, by those of Braidwood and Vacher. || 
 
 These investigations were followed in 1873-74 by those of 
 Letzerich, which are spoken of by almost all later writers. § 
 
 In the work of Cornil and Babes on Bacteriology, we find on 
 this subject only the following short notice : — " He [Letzerich] has 
 endeavoured to find and to cultivate the microbes in the sputa of 
 Whooping-cough ; and has described and figured some enormous 
 micrococci, which dispose themselves in irregular chains and in 
 zooglffia. He even professes to have produced a convulsive cough 
 
 * Poulet, Comftes-rendus de PAcad. des Sciences, 1867, Lxv,, p. 254 ; 
 Presse Mklicale Beige, 1867, No. 51. In the original Italian edition I over- 
 looked the fact that in 1867 Dr. Cerasi, before the investigations of Poulet were 
 made public, found in the sputa of Whooping-cough, under a power of 150 
 diameters, certain forms of fungi, similar to those described later by Letzerich, 
 and which he called " Oidium pertussis." Nevertheless, these undoubtedly 
 anterior researches escaped the notice of authors, who turned their attention to 
 the sputa of Whooping-cough. — Cerasi, Giornale delle Scienze Mediche di 
 Venezia, Serie III. , Tomo XII. (F. V. ) 
 
 t Gerhard t, Tratt. conipleto delle Malattie dei Bambini, Versione Ital., Napoli, 
 1883, Vol. II., p. 474. 
 
 + A. Ransome, _/o«rK. Anat. and Physiology, 1870, iv., p. 217; also 
 Proc. Lit. and Phil. Soc. , Manchester, 1 870, Vol. ix. p. 106. 
 
 II Braidwood & Vacher, Researches on Measles, Trans. Path. Soc, London, 
 1878, XXIX., p. 422. 
 § Virchow's Archiv., 1873, Vol. Lvii., p. 518, and 1874, Vol. LX., p 409. 
 
4 SPUTA OF WHOOPING-COUGH. 
 
 in rabbits by inoculating them, by tracheotomy, with micrococci. 
 This work of Letzerich is absolutely contradicted by a communica- 
 tion from Biirger,* who finds in the sputa of Whooping-cough many 
 small, rod-like, ellipsoid bodies, with a restriction in the middle. 
 The sputa, to the naked eye, resembles tufts of matter, whitish at 
 first, afterwards turning slightly yellow. It is these tufts, spread 
 upon the glass slides, dried, stained with methyl violet, and 
 bleached by alcohol, that show the rod-like bodies. They are 
 found in or on the cellules, chain-like bodies being entirely absent. 
 Biirger thinks that the drawings of Letzerich do not represent 
 these parasites. According to Biirger, these rod-like bodies, so 
 numerous in Whooping-cough, bear an intimate relation to the 
 disease and its stages. He never cultivated them. As these 
 micro-organisms can so easily be accidentally present both in the 
 mouth and pharynx, they must always be looked upon with 
 suspicion." t 
 
 Hagenbach gives a more precise account of the work of 
 Letzerich in the monograph cited above. On the subject of the 
 sputa in Whooping-cough, he quotes : — " During the period of 
 convulsions we find in the mucus — fungoid filaments — sometimes in 
 very great quantities, interwoven and ramified, and in which a 
 vigorous formation of spores takes place. 
 
 " This fungus, a specific sign of the disease, can be detected 
 with the naked eye. It is not reproductive in the mucus mem- 
 brane, but lodged in the corpuscles of the mucus, and not on the 
 epithelial cells. The course, duration, as well as the intensity of 
 the paroxysms of the cough, vary with the fructification or propa- 
 gation of this fungus, producing spasmodic irritation in the mucus 
 membrane. The introduction of these fungi into the larynx or 
 tracheae of rabbits will in six to eight days bring about spasms of 
 convulsive coughing. As a rule, the fungus of Whooping-cough 
 invests the folds and cryptse of the epiglottis, the larynx, and the 
 tracheae, finding its way even into the air-cells of the lungs, giving 
 rise in its growth to catarrhal and inflammatory symptoms." % 
 
 * Berliner klinische Wochenschrift, No. i, 1883. 
 t Cornil et Babes, Les BacUries et leur rtle dans FAnat. et FHist. Path, 
 des Malad. Infec. Paris, 1885, pp. 556, 557. 
 
 + Gerhardt, loc. cit. , p. 474. 
 
SPUTA OF WHOOPING-COUGH. 5 
 
 Further on :— " Letzerich has found the smaller bronchi of the 
 inoculated rabbits, and even the air-cells, blocked by them, mixed 
 with micrococci and a slender mycelium." * 
 
 In the fifth section relating to the fungi, which I have found in 
 some specimens of sputa and their fructification, we shall have an 
 opportunity of again referring to this. 
 
 Following the investigations of Letzerich, Henke in the same 
 year (1874) discovered in the sputa, pus, corpuscles, and other 
 round bodies (? young epithelial cells), full of minute particles in 
 most lively motion, and which could be paralysed by solutions of 
 quinine. + To myself, it seems very probable that this motion was 
 not at all due to the corpuscles themselves, but was such a vibra- 
 tory motion as is seen in all living cells, whether of the saliva, or 
 white corpuscles of the blood, pus, mucus, or young ' epithelium 
 cells, wherever an osmosis takes place. Very striking examples of 
 such motion is seen in the corpuscles of the saliva, of mucus, of 
 normal sputa, of nasal mucus, or, better still, of the mucus of the 
 urethra, by allowing water, pure or coloured with aniline, to run 
 into the fresh preparations. 
 
 Tschamer, of Gratz, in his many researches during the year 
 1870, had not observed, except in a very few cases, any microbes 
 in the sputa of Whooping-cough ; and then only such as are found 
 in healthy sputa, especially that expectorated in the morning. But in 
 1874, one or two days before the convulsions set in, he came 
 across a reticulated mycelium, and many round or ovoid spores of 
 different sizes. These were at first colourless, but subsequently 
 became yellowish or of a darkish red colour. He thought these 
 to be very similar to, if not identical with, the dark mould on 
 decaying orange or lemon peel, and looked upon the spores of this 
 mould as the germs, or the pathogenic cause, of Whooping-cough. 
 
 The observations of Tschamer were confirmed by Oltramare 
 and others, but contradicted by those of Rossbach of Wiirtzburg. 
 But more of this in our fifth section. % 
 
 In the year 1883, Burger of Bonn describes, as we have seen 
 
 " Ibidem, p. 485. 
 
 \ Arch, fur Klinische Medicin, XII., 1874, p. 630. 
 
 J Barlow, loc. cit., p. 916. 
 
6 SPUTA OF WHOOPING-COUGH. 
 
 in the extract from Cornil and Babes, the microbes in the sputa 
 of Whooping-cough as sometimes linking themselves in chain-like 
 bodies, the larger ones being constricted in the middle and others 
 forming clusters in colonies. He took them to be quite distinct 
 from those of the Leptothrix.* 
 
 In the same year (1883) Moncorvo, of Rio de Janeiro, 
 observed swarms of micrococci scattered about the mucus or 
 inclosed in the pus corpuscles or in the epithelium cells, which 
 seemed to him to gradually decrease or disappear altogether, as 
 the disease ran out its course. Local applications of Resorcina 
 paralysed the movements of these micrococci.t 
 
 Full particulars of these researches from those of Tschamer to 
 those of Moncorvo will be found in Barlow's -papers. 
 
 In the year 1886, at the end of his investigations, Barlow pub- 
 lished the results of his microscopical examinations of the sputa in 
 Whooping-cough — examinations undertaken with the help of 
 Broadbent. A i/i2th inch objective was used, magnifying to 800 
 diameters. The preparations were stained with methyl violet and 
 showed numbers of nuclei of pus corpuscles, fibrinous particles, 
 and a few epithelium cells, some covered with minute micrococci 
 in chain-like lengths or masses (zooglaea). Though similar micro- 
 cocci were to be seen all through the rest of the field, nevertheless 
 it was evident that the principal nidus was in the epithelium cells. 
 Of this remarkable fact I shall have more to say. From these 
 observations Barlow was led to believe that the cause of Whoop- 
 ing-cough was to be found in the disquamation of the epithelium 
 cells of the larynx, brought about by foreign growths. J 
 
 Michael of Hamburg, in the same year 1886, declared his firm 
 opinion to be that Whooping-cough arose from a reflex and spasr 
 modic action, partially of the pneumogastric nerves, but principally 
 from that of the muscles of the upper part of the larynx, caused 
 by a specific irritation of the nasal mucus membrane, produced 
 by certain microbes. On this account he recommended nasal 
 
 * Berliner Klinische Wochenschrift, 1883, No. I. 
 
 t Moncorvo, De la Nature de la Coqueluche et de son traitement far la 
 Resorcine. Rio de Janeiro, 1883. Archiv. di Patalogia Infantile. Napoli, '85. 
 
 X Barlow, loc, cit., p. 967. 
 
SPUTA OF WHOOPING-COUGH. 7 
 
 insufflation of gum benzoin, quinine, and nitrate of silver mixed 
 with calcined magnesia.* 
 
 Hack, Schadewald, Wille, and Sonnenberger took the same 
 view, as may be seen from an article in the above-mentioned 
 periodical,^ and from a short notice of it by Morgagni in the 
 same year. J 
 
 But these writers do not speak of the morphology or biology of 
 these nasal parasites, nor of the methods of staining them for 
 observation. Nor is it clear that their opinions are derived from 
 their own bacterioscopical examinations, or are simply hypotheses 
 put forward to uphold particular therapeutical theories, or to 
 explain the true or supposed virtue of certain local remedies as 
 those above mentioned, or others, such as iodiform, salycilic acid, 
 boracic acid, injections or inhalation of corrosive sublimate, tinc- 
 ture of iodine, and alum, || or the use of other local or general 
 applications of quinine, antipyrin, etc.§ 
 
 Until more is known of this parasite, I think it best to leave 
 its discussion, turning my attention more exclusively to the exami- 
 nation of the sputa in Whooping-cough. My conclusions, as will 
 appear, are not opposed to the idea of nervous spasmodic action, 
 and are compatible with other researches as to the seat of its first 
 origin. 
 
 The following notice in the Lancet of December, 1887, will 
 show results of the work of Afanasieff : — 
 
 " Doctor Afanasieff has succeeded in finding and cultivating 
 what he believes to be the true bacillus of Whooping-coiigh. This 
 microbe differs distinctly from all other bacteria which have been 
 described. It is somewhat like Friedlander's Pneumonia bacillus, 
 but is shorter and thinner than the latter ; besides, in gelatine it 
 
 * Michael, Deutsche Medicinische Wockenschrift, 1886, No. 5, p. 74. 
 t Sonnenberger, same article. 1887, No. 73. 
 ni, 1888, Part II., No. 20, p. 256, and No. 39, p. 473. 
 
 II Vide, among others, Nasal Treatment of IVhooping-cough, Lancet, 1887, 
 Vol. I., p. 136. 
 
 § As, for example, Deutsche Medecinische Wochenschrift and Morgagtzi in 
 articles cited above. 
 
8 SPUTA OF WHOOPING-COUGH. 
 
 does not form nail-shaped cultures, those which are produced 
 having no hemispherical head.* 
 
 " Its potato cultures too, are quite different from those obtained 
 from Friedlander's bacillus. Afanasieff's bacillus exhibits a re- 
 markable degree of vitality, for jelly cultures that have become dry 
 and have been kept for four months, appearing under the micro- 
 scope to be more or less destroyed, are still capable of producing 
 fresh cultures when fresh media are inoculated from the dried mass. 
 " Dr. AfanasiefPs researches were chiefly made from the sputum 
 of some of his own children who were affected with Whooping-cough. 
 The mouth was well washed out with a permanganate solution, 
 and the mucus coughed up afterthe next paroxysm or two examined. 
 In this mucus, after staining with methyl-violet, and in the pus- 
 corpuscles contained in it, the bacilli could be seen with a magni- 
 fying power of from 700 to 1,000 (Zeiss's eye-piece 3 or 4, 1/12 
 inch oil immersion objective) as short rods, sometimes single, 
 sometimes in twos, or even in short chains running in the direction 
 of the mucus, sometimes again in small cultures. Their length 
 was from o'G fi io 2-2 fi. Of course other bacteria were found. 
 Pure cultures, however, were easily made on agar-agar, meat 
 peptone, jelly, potato, etc. Dogs and rabbits were inoculated with 
 a fluid culture, mixed chloride of sodium solution, some by means 
 of injections into the trachea, others by direct injections into the 
 lungs. All the animals were seriously affected, and many of them 
 died. The symptoms were somewhat similar to those of Whooping- 
 cough, including cough, dyspnoea, and redness of the eyes. Many 
 of the cases were complicated by broncho-pneumonia. On ex- 
 amining the bodies of those animals which died, the mucus 
 membrane of the air passages were found much reddened, and 
 coated with a tenacious clear mucus, in which, as well as in the 
 pneumonic patches in the lungs, the bacilli were found. Similar 
 appearances and the same bacilli were observed in the bodies of 
 children who had died from Whooping-cough. As to treatment, 
 inhalations, and spray of various antiseptic drugs would appear 
 
 * Dr. Afanasieff has personaljy examined and cultivated the so called 
 miciococcT. ( Compes Rendus hebd. SocUU de Biohgie, May 21, 1884, p. 356). 
 As to the club-headed cultures, see among others Cornil and Babes, loc. cit., 
 PI. xxvu., Fig. 27 (F. V.) 
 
SPUTA OF WHOOPING-COUGH. 9 
 
 to afford the most ground for hope in relieving and shortening this 
 complaint. The author points out that the quinine, benzoin, and 
 other substances which have been used as applications to the usual 
 mucus membrane by Michael and others, under the idea of com- 
 bating a reflex affection, are really, perhaps, beneficial from their 
 property of destroying bacilli." * 
 
 The Medical Times of Philadelphia, of April, 1888, gives a 
 shorter notice of AfanasiefFs discoveries.! 
 
 § 11. 
 
 A SHORT ACCOUNT OF CASES, IN WHICH THE SPUTA 
 HAVE BEEN EXAMINED. 
 
 The thought of examining the Sputa in cases of Whooping- 
 cough did not occur to me till the epidemic of this year, when I 
 came across a truly typical case of Whooping-case, typical both in 
 its clinical symptoms and in the character of the Sputa. The 
 principal features I have found in Sputa of Whooping-cough, as 
 will be seen, were the presence of an extraordinary and peculiar 
 abundance of masses, particles, filaments, and granules of myelin, and 
 it was through observing the quantity of myelin that caused my 
 surprise in this first case, a quantity that took up quite one half of 
 the surface of the preparations examined. Perhaps if I liad not 
 come across such a striking case, I should not have given so much 
 importance to the presence of myelin nor renewed my researches 
 in other cases. The microscopical examinations were undertaken 
 on this occasion through a special interest in the little sufferer and 
 the fear of some other threatening complication. 
 
 First Case. 
 
 A.S., a boy of 8 years, plethoric, robust, and well developed 
 for his age, was in the latter part of March seized by attack of 
 measles with a distinct nasal catarrh, confluent eruption, somewhat 
 haemorrhagic. After the disquamation, he showed symptoms of 
 
 * The bacillus of Whooping-cough.— Lancet, Dec. 3, 1887.— Vol. ii, p. 1131. 
 t Philadelphia Medical Times. Aprils, iSSS.—'No. Jjo., p. 403. 
 
10 SPUTA OF WHOOPING-COUGH. 
 
 bronchial catarrh, which, though slight at first, became more dis- 
 tressing by the middle of April, and at last took the form of 
 Whooping-cough. 
 
 The boy was subject to some twelve paroxysms per hour, with 
 facial adema, ecchymosis of conjunctive membrane, epistaxis, 
 and a severe form of quintes, as the French call them. He had 
 no signs of capillary bronchitis or pneumonia, nor any fever. The 
 case was therefore truly typical in the form of the paroxysms and 
 in the absence of other complications. 
 
 Sputa of May 10. — (a) The first sputum taken for examination 
 was after the third week of the period of convulsions of the 
 Whooping-cough ; it was colourless. Hyaline, very viscid, and 
 found to be free from any residuary particles of food. It contained 
 but few pus corpuscles, the whole mass being composed almost 
 entirely of the elements which will be described. No red blood- 
 corpuscles were found. A fair quantity of buccal epithelium cells 
 and pavement epithelium cells from the air-passages. I thought it 
 would be superfluous to reproduce these in the plates, with the 
 exception of a small particle of buccal epitheUum cell, as in f. 
 Fig. 2. In Fig. i, >^ is a salivary corpuscle, stained with gentian 
 violet, magnified to 1,750 diameters. Vibratile epithelium cells 
 from the air-passages were to be seen ; a small one is shown in f. 
 Fig. I, coloured in the same way and magnified to 2,500 diameters. 
 Rather a moderate number of ellipsoidal cells of epithelium were 
 present from the air-passages, called sometimes alveolar epithelium 
 cells. This is not the place to discuss whether or lio these cells 
 truly come from the air-passages or from the air-cells.+ One thing 
 is certain — that they are found often covered with pigment grains 
 and with myelin in sputa of all kinds. I have found them in 
 those of simple hoarseness. 
 
 In Fig. I, g, one of these cells will be seen, in its natural state, 
 with the granules of myelin magnified to 400 diameters. 
 
 From the very first, I was greatly surprised to find that nearly 
 
 * Epistaxis — bleeding from the nose. 
 
 t Senator held them to be epithelium cells from lower parts of the bronchial 
 mucus membrane. (Bizzozero, Man. di Micr. Clinica, Milano, 1882 
 p. 1882, pp. 144—145.) 
 
SPUTA OF WHOOPING-COUGH. 11 
 
 the half of the surface of the preparations consisted of numerous 
 masses of myelin, in all sizes and shapes. Fig. i, d, shows one 
 rather smaller than others, drawn in its natural state, magnified to 
 400 diameters. Among these masses were seen myriads of parti- 
 cles of myelin, some isolated, some grouped together, as shown in 
 a, a, a, a; others are shown in b, b, b, b, b (stained with gentian 
 violet), both magnified to 1,000 diameters. Besides these, there 
 were numberless grains of myelin smaller than the most minute 
 bacteria and barely visible, unstained, but rendered quite distinct 
 by gentian violet. These are shown in b on the left of the grouped 
 particles, and magnified to 1,000 diameters, but better still in 
 Fig. 2, i, where they are magnified to 2,500 diameters. Sorne 
 portions and detached filament are shown in Fig. i, b (1,000 diam.), 
 and at i are shown, for comparison, some minute particles and 
 some of the smaller granules unstained, as given by Dr. Beale, 
 magnified 2,800 times.* 
 
 It wjas evident that the viscosity and hyaline character of the 
 sputa was due to the great amount of myelin present, but of this 
 more later on, as well as of the bacteria and bacilli found in the 
 sputa. It will be enough now to say that both in the sputa of this 
 case and subsequent ones, there were found, even when fresh, 
 many forms of spirilli, Lepiothrix buccalis, Vibrio, Bacillus 
 subtilis, which increase day by day with the composition of the 
 mass. Too many plates and too much space would be required 
 to describe singly all the forms and varieties of the bacteria and 
 bacilli. Therefore, only the principal forms will be given in the 
 illustrations. 
 
 The different bacteria and bacilli of Whooping-cough are 
 stained with gentian violet. Fig. 2 shows a group of minute bac- 
 teria in a (800 diam.), some of which are reproduced in b (2,500 
 diam.). In c will be seen two chain-like bacteria— one shorter and 
 twisted, the other longer (800 diam.) ; d shows a part of the 
 longer chain, magnified to 2,500 diameters. Two dumb-bell bac-. 
 teria are shown in e (2,500 diameters), one of which is quite cap- 
 sular in appearance. Of such dumb-bell bacteria there was great 
 abundance, some spread throughout the mucus mass, but most of 
 
 * Beale, The Microscope in Medicine, London, 1878, PI. xxi., Fig. 6. 
 
12 SPUTA OF WHOOPING-COUGH. 
 
 them swarming around buccal epithelium cells, specially around 
 those showing signs of disintegration or corrosion. In / will be 
 seen a portion of buccal epithelium cell invaded by both these 
 forms of bacteria under the power of 2,500 diameters. 
 
 Such dumb-bell bacteria formed the greater part of the bacteria 
 of the sputum, seemingly identical with those discovered by Poulet 
 in vitiated air and by Biirger in sputa. 
 
 Some extremely minute bacteria and bacilli are shown in g 
 (2,500 diameters), with two specimens of curved diplococci mor- 
 phologically identical with the so-called gonococci of Neisser ; but 
 these curved diplococci were only found in some few portions. 
 
 In h is reproduced (2,500 diameters) a corpuscle shaped after 
 the pneumococcus of Friedlander, but larger, and which might have 
 been a very small pus corpuscle misshapened, as shall be men- 
 tioned later. 
 
 In the first examination but few specimens of the forms /, k, 
 and i were observed. Few also of those interwoven, as in u and 
 u, were seen. These last appeared afterwards in greater numbers 
 in this same sputa and also in other sputa. 
 
 Divers kinds of bacilli also were noticed. Two are shown in 
 m, m (2,500 diameters), which seem to be more mature forms of 
 the young bacilli, seen in n and «, which were found in other cases. 
 
 These examinations of bacteria and bacilli were made with a 
 , homogeneous immersion lens, i/i8th inch, and a Dujardin con- 
 denser of the improved pattern, made by Messrs. Bezu, Hausser, 
 and Co., successors to Hartnack and Prazmowski. The drawings 
 have been faithfully made from the preparations. 
 
 (b.) The day after. May n, other portions of the same 
 sputum were examined and found to contain the above forms of 
 microbes, in increased numbers, specially the beaded strings, as 
 in i and k. The forms shown in / (2,500 diam.) were observed, of 
 which some may represent smaller specimens of the forms d 
 and If, others are bacteria, very small and globular or oval in 
 germination. From these oval forms may be derived those of a 
 long biscuit shape ; of these the greater part were capsular in 
 appearance. They are seen low down and on the left in the figure, 
 some in twos, some in chain-like lengths, generally grouped round 
 the buccal epithelium cells or penetrating the cells themselves. 
 
SPUTA OF WHOOPING-COUGH. 13 
 
 Other bacteria of this group are, instead, representatives of the 
 same forms d and e, increased in size, with their heads not yet 
 united. Many individuals have a capsular appearance as seen on 
 upper part and to the right of the figure, at L There seems to me 
 every reason to believe the diplococci, though smaller, to be similar 
 to the pneumococcus of Friedlander, of which Dr. Afanasieff gives 
 an account. It may be here observed that I have found analogous 
 or identical forms in sputa under other circumstances, in normal 
 sputa and in sputa during affections of the air passages, likewise 
 in saliva, nasal mucus, urine, and even in spermatic fluid. 
 
 On the left and above the figure being described there is a 
 curved diplococcus, similar to that in g, but somewhat larger ; in 
 the chain (/, p. Fig. 2) there are some still larger, and the similarity 
 naturally suggests that the drawings g, /, /, are but three degrees 
 of development and increase of one and the same bacterium. 
 
 It was, perhaps, by inadvertence, that no trace of fungi was 
 noticed the day before, but on the eleventh I observed germinating 
 filaments of fungi of fair size. They were, at first, all after the same 
 type, as in Fig. 3, d, where they are shown stained with gentian 
 violet and magnified to 800 diameters. It should be said that 
 while the figure shows them with a regular elliptical outline, as in 
 e,f, g, etc., they were in reality somewhat flattened at- the ends. 
 
 Not to anticipate the description, given later on, of this fungus, 
 I would merely state that these filaments became more conspicuous 
 during the following days, and a great many vescicular gemmules, 
 sacculi, or sporidia were noticed, some yet attached to the filaments, 
 others already fallen off As these closely resemble those spoken 
 of by Hagenbach as observed by Letzerich used in his inoculation, 
 they are given in Fig. 3, b, b, stained with gentian violet, and in c, 
 with carmine [Fig. y is printed in violet, to be uniform with Fig. 
 T,bh\, magnified to 2,500 diameters. 
 
 (c.) During the following days there appeared in the sputa 
 (which had been kept in a tightly covered porcelain vessel), among 
 the larger filaments described above, other smaller ones, with many 
 small rounded spores of a light greenish colour. 
 
 In the same interval the larger filaments had increased in 
 numbers to such a point that by the i6th (the sputum was then 
 five days old) one could count some twenty or thirty filaments both 
 
14 SPUTA OF WHOOPING-COUGH. 
 
 small and large in one single preparation. It was probably due to 
 the multiplication of the greenish spores that the mass of the 
 sputum took the same greenish colour and gave off a musty smell, 
 which increased till the whole dried up in about a month at a 
 temperature of 20° or 22° C. 
 
 It was during this time that I observed the fructification of 
 both these fungi. That of the larger ones, as the filament, d, is 
 shown in Fig. 3 k and ^ (140 diameters) ; that of the smaller ones, 
 the filaments of which are not shown, is given in n. 0. p. 
 (800 diameters). Spores detached in the fructification of the 
 first fungus will be seen in / and /, those of the second in m and m 
 (t,ooo diameters). 
 
 I intend to refer later on to my observations on sputa after 
 boiling and on the dust deposited in the room of the sick boy. 
 
 These few notes will suifice for the present in regard to the 
 sputum of May 10. 
 
 Sputum of May 12. — On this day another portion of the sputum 
 was taken and it gave the same results as that of the day before, 
 showing the same extraordinary amount of myelin. In these were 
 present in great quantities the chain-like or beaded bodies budding 
 at one extremity as in Fig. 2. /, or at both ends as in k. These 
 beaded forms have been found in other cases of Whooping-cough, 
 in other sputa, in saliva and various other substances. 
 
 On the second day large numbers of the filaments of fungi 
 appeared in this sputum, as before described. I have not had the 
 leisure to follow the development of the filaments, nor to ascertain 
 whether the smaller ones made their appearance. 
 
 Sputum of June 5. — It was during the seventh week of the 
 convulsions that an attempt was made to ascertain if the same 
 amount of myelin was still to be found, and it was observed in the 
 same large quantity. No great notice was taken of the bacteria in 
 this sputum, but the large filaments of fungi and their germina- 
 tion were noticed, as in the preceding cases. 
 
 Towards the end of June, the cough became less intense and 
 less frequent, and by the end of this month might be said to have 
 passed away ; but the boy still brought up from time to time a 
 yellowish mucus, such as is seen in a case of very mild catarrh. 
 
 But the clinical history of the case does not end here. The boy 
 
SPUTA OF WHOOPING-COUGH. 15 
 
 on the 28th of July, through his profuse perspiring, was seized 
 with an attack of tonsillitis, and was slightly feverish. Two days 
 afterwards, on the 30th, at lo a.m., he was in a high fever, subject 
 to shivering fits, with heavy breathing and a sharp pain on his 
 right side. By the evening his pulse was 160 and his temperature 
 40-1° C. During the night some of the expectorations were 
 tinged with red. In the following morning (the 31st July) a fresh 
 portion of sputum was taken for examination under the micro- 
 scope. 
 
 Pneumonitic Sputum of July jist. — This sputum was somewhat 
 tinged with red, and on being examined was found to contain a 
 large number of red blood-corpuscles, a fair quantity of ellipsoidal 
 epithelium cells, covered with small granules of myelin ; through- 
 out the mass some few smaller particles of myelin were observed, 
 which seemed ^ to have fallen away from the ellipsoidal epithelium 
 cells. But no vestige was to be seen of the innumerable particles 
 nor of the numerous filaments and masses of myelin which had 
 been found in such large quantities before in the sputa during the 
 Whooping-cough. 
 
 What had become of the extraordinary abundance of myelin ? 
 If the copious formation and elimination of this matter arose 
 from some special condition or idiosyncrasy of the subject, there 
 would be no reason why they should not appear in this lung 
 attack ; all the more, as the quantity of sputum from the lungs in 
 twenty-four hours was very moderate. So that an equal amount 
 of myelin in this sputum would have given a much higher per 
 centage than that shown during the Whooping-cough. But this 
 matter will be taken up later. 
 
 The bacteria found in this sputum were not in any way differ- 
 ent from those that are usually met with in other pulmonary 
 attacks, and, on the whole, were not unlike the bacteria of sputum 
 in cases of Whooping-cough. The forms e, f, and /' (Fig. 2), 
 with a capsular appearance, were rather abundant. Many forms 
 in couples and in chains were present ; they are shown in/,/, and 
 are met with in cases of pulmonary complaints. Many of them 
 had a capsulated appearance, sometimes enclosing two in the 
 same transparent envelope, as in /', or in threes and fours, and in 
 lengths of six or more. All the forms e,f, and /', as well as the 
 
16 SPUTA OF WHOOPING-COUGH. 
 
 forms /, /, and / ', were seen either covering or penetrating the 
 buccal epithelium cells. 
 
 It should be noticed that the larger bacteria, as in / and /', 
 are found — and in great quantities, too, at times — in the sputa of 
 whooping and other sputa. But the author never met with the 
 forms curved after the fashion of the so-railed gonococci intermixed 
 in the chains except in Whooping-cough ; nor observed these 
 forms having in Whooping-cough a capsular appearance. 
 
 Besides the above, the forms shown in /', to the right, and in 
 / under the figure were plentiful ; forms which, together with that 
 in/', may be classed with so-called pneumococci. They are cer- 
 tainly numerous in the sputa in cases of pneumonia and pleurisy 
 cases, and are to be found generally in other sputa and other 
 substances. 
 
 It was very easily noticed in this specimen of the sputum how 
 the forms /' and /', with the knob-heads separated, passed into 
 those at /' with the knobs as yet only partly joined, and into those 
 at e and /all in one, and all the more, as these last two forms are 
 the same as those with the knobs separated, only seen fore- 
 shortened. 
 
 Along the striae of the mucus were seen swarms of very minute 
 bacteria; but they were similar in form to those shown in /and/, 
 and may be thought to be only younger specimens of the same type. 
 
 During the following day — August ist — a greatly increased 
 number of these minute bacteria were found in other preparations 
 of the same sputum, and others more minute were observed in 
 such beaded shapes, as shown in Fig. 2, s, and in interwoven 
 masses, as in u and u. The first are magnified to 2,500 diameters ; 
 the second, for the sake of space, only to 600 diameters. These 
 forms are found in other sputa and will be further described. 
 Among the swarms of the minute bacteria, I observed some 
 slender bacilli in their first growth (see b\ 2,500 diam.), where will 
 also be seen some very small bodies, perhaps micrococci, which 
 appear to be in process of being cemented together by a finely 
 granular matter forming between them. The chain-like bodies, as 
 in c, were present, and more than ever the forms p and / ', which 
 appears still more evidently to force their way into the buccal 
 epithelium cells. 
 
SPUTA OF WHOOPING-COUGH. 17 
 
 The forms shown in b', /, «', //, /,' on the left, /, /, lower 
 down, and in x' are stained with methyl violet, and are from cases 
 of mflammation of the lungs or pleurisy ; it is my experience 
 that methyl violet in these cases renders the capsular appearance 
 more distinct. 
 
 I shall have more to say on the sputa of Pneumonia or 
 Pleurisy ; as regards the boy the disease soon ran its course, with 
 a morning temperature of 39°C., 39-4°C., and evening tem- 
 perature of about 40° C. The fever abated on the fifth and the 
 boy was well again by the twelfth of August. 
 
 Naturally the results of the examination of the sputa in the 
 case of this boy led me to further researches in other cases of 
 Whooping-cough. In this there was a certain difficulty, for in the 
 case of infants the expectoration is usually very limited, only small 
 portions can be obtained, and in these the very elements sought for 
 may easily be wanting. For myelin is not always diffused to 
 the same,amount in sputa from different parts of the bronchial 
 secretion. And then, again, in private and ordinary practice it is 
 not always easy or becoming at times to seek for material for 
 examinations or researches that do not tend to the immediate 
 relief of the patient, _and in fact I could only in fourteen other 
 cases find means of examining the sputa. These cases will be 
 considered in the following pages. 
 
 Second Case. 
 
 On the 1 8th of May I examined the sputa of a little girl aged 
 five years during the seventh week of the convulsions. The 
 sputum was to all appearance a puriform mucus. Red. blood- 
 corpuscles were to be seen, but in no great quantity ; a fair quan- 
 tity of pus corpuscles; a few ellipsoidal epithelium cells were 
 present, together with many buccal epithelium and pavement 
 cells from the air-passages, but very few of the small vibratile cells 
 from the same. 
 
 Very few particles of myelin were found here and there in 
 the mass. However, the day after, the sputa having been kept in 
 a closed tube, the myelin, being a lighter body, came to the 
 surface in such quantity as to form about one-tenth of the whole 
 quantity. This habit that myelin has of rising to the surface of 
 sputa will be alluded to again. 
 
18 SPUTA OF WHOOPING-COUGH. 
 
 The bacteria were much of the same forms as in the preceding 
 case, but in far greater quantity, being found principally upon the 
 buccal epithelium cells. 
 
 Third Case. 
 
 The sputa of an infant of ten months was examined on the 
 19th of May, during the first week of the convulsions. Several 
 red blood-corpuscles were observed, many pus corpuscles, very few 
 ellipsoidal cells of epithelium, and fewer vibratile cells. Buccal 
 and pavement cells of epithelium were present as before. Very 
 few granules of myelin were found, and the bacteria were much 
 the same as those observed in the first case. A good many speci- 
 mens of the form 0, Fig. 2 on the left ( x 2,500 diara.), presented 
 themselves. This form, as can be seen from the figure, is much 
 the same as that in g and / ; in this case, there appears something 
 of the nature of a cementing medium between the cocci. Some 
 minute diplococci, with well-defined capsules feebly showing the 
 darker cocci within, are represented in (upper part of Fig. 2). 
 There were other more minute bacteria, but of the same form as 
 those before described. The infant died during an attack of 
 convulsions. 
 
 Fourth Case. 
 
 I was able on the 20th of May to examine the sputa of a boy 
 two and a-half years old during the first week of the convulsions. 
 The same morphological characters were found, and myelin was in 
 the same small quantity. The bacteria were the same as those of 
 the first case ; but a number of bacilli appeared, some longer, 
 some shorter, showing internally signs of segmentation, granular 
 masses or bacteria, or mere specks of coloured matter (see n, 
 Fig. 2, magnified 2,500 diameters). There will be occasion for 
 dwelling on the similarity of this form and that in m, m, referred 
 to in the first case, as well as on the probability of the origin of 
 the larger bacillus, n, from the mass, «, u, and that of the smaller 
 bacillus, shown close by, from that of the beaded form, v. 
 
 Among my notes there is no mention of fungi in these three 
 last cases, but perhaps they may simply have been overlooked. 
 
 Fifth Case. 
 On May 24 the sputa were examined in the case of an infant 
 
SPUTA OF WHOOPING-COUGH. 19 
 
 of five months during the third and fourth weeks of convulsions, 
 in which the quinte was very pronounced. The sputum was dense 
 and so thick, that in order to spread a portion on the slide it was 
 necessary to dilute it with a drop of chloride of sodium solution. 
 Red blood-corpuscles were absent, but a few pus-corpuscles. 
 Many ellipsoidal epithelium cells, some few vibratile epithelium 
 cells, with the usual pavement cells, were found. Myelin was in 
 almost the same abundance as in the first case. The smaller par- 
 ticles and lighter portions could be observed to collect on the 
 edges of the preparation, enclosing within the liquid belt the 
 thicker and viscid material. The bacteria were very similar to 
 those in the first case. 
 
 On the following day, the germinating filaments of fungi and 
 the vesicular gemmules of the first case were again met with in 
 another portion of the same sputum. 
 
 This infant recovered from the Whooping-cough, but died 
 three months afterwards from inflammation of the right lung. 
 
 Sixth Cask. 
 
 The sputum of a boy four years old was examined on the 24th 
 ' May during the fourth week of the convulsions. The attack was 
 of a much milder form and the cough less frequent than in the 
 preceding case. Here, too, the sputum was dense and thick, con- 
 taining some few red blood-corpuscles, a few pus-corpuscles, 
 scarcely any ellipsoidal epithelium cells, but about the same 
 amount of the pavement and vibratile epithelium cells as in the 
 case above. Myelin was present in a fairly abundant, although 
 not in such an- enormous quantity as seen before. The forms of 
 bacteria,/ and/, were observed, besides those described above. 
 One bacillus was seen slightly coloured, enclosing some darker 
 bodies, small masses— perhaps bacteria— irregularly disposed ; it 
 is shown in n on the right and magnified 2,500 diameters. 
 
 It appeared to me to be a younger specimen of the bacillus 
 m m, met with before. 
 
 Seventh Case. 
 
 {a) On May 2Sth I examined the sputum of a boy two years 
 old during the second week of the convulsions. The cough was 
 not so very distinct from that of an ordinary cold, but harsh, and 
 
20 
 
 SPUTA OF WHOOPING-COUGH. 
 
 taking into consideration that other boys of the family were evi- 
 dently suffering from Whooping-cough, I considered it to be of a 
 distinctly specific character. The boy had had a fit of convulsions 
 two days before, following a severe attack of indigestion. In the 
 stool of the lad portions of orange-peel were found. He was 
 teething and feverish. 
 
 In the sputum examined red blood-corpuscles were abundant, 
 as also pus-corpuscles and the ellipsoidal epithehum cells ; but of 
 myelin little or nothing could be seen. 
 
 (d) The sputa were again examined on the 5th of June during 
 the fourth week of the attack, which was still of a rather dubious 
 character. The fever had disappeared. I found a good number 
 of particles and granules of myelin, many masses and many iso- 
 lated specimens of the beaded bacteria (Fig. z, s, magnified 2,500 
 diameters). These bacteria, in form, size, and disposition, were 
 very similar to the so-called Koch's bacillus or tubercular bacillus, 
 but in a dried preparation they were completely bleached by a 
 mixture of nitric acid and alcohol (i 13). Many bacteria were 
 observed similar to those in p, p, which, as well as those beaded 
 forms, s, were principally crowding about or within the buccal 
 epithelium cells, as has been before mentioned. Here, too, were 
 observed the larger bacilli of the form, as in m, m. 
 
 Eighth Case. 
 (a) On the 27th of May the sputum of a little girl aged seven 
 was examined during the fifth or sixth weeks of convulsions. 
 The girl had some slight and irregular attacks of fever, but every 
 symptom of either capillary bronchitis or catarrhal pneumonia 
 were altogether wanting. Red blood-corpuscles, pus-corpuscles, 
 and ellipsoidal epithelium cells were present. Myelin was found 
 in considerable quantity ; the particles and free granules were so 
 numerous in proportion to the few epithehum cells, that they 
 could not be said to be simply a diffusion of the myelin so fre- 
 quently found clothing such cells. Among other forms of bacteria, 
 those shown in /, p, were in extraordinary numbers, some arranged 
 in line, others united together in twos and fours. Of those in line 
 with one another, some seemed bent and to have a striking 
 likeness to gonococci, as in the figure drawn from this very prepara- 
 
SPUTA OF WHOOPING-COUGH. 21 
 
 tion. In Fig. 2, r, is shown a large bacillus with four articulations, 
 and in ^ a form with one articulation. Both were observed in 
 this sputum (magnified 2,500 diameters). 
 
 (p) On the 9th of June, the eighth week of the con- 
 vulsions, the sputum was again examined, but little or no 
 myelin was found. In this sputum the bacteria in the largest 
 numbers were the groups, isolated specimens, or beaded chains 
 shown in d, e, and/ 
 
 Ninth Case. 
 On the 13th of June I examined the sputa of a little girl 
 SIX years of age, during the fourth week of the convulsions. 
 Some few red blood corpuscles were found, a good many pus 
 corpuscles, only a few ellipsoidal cells of epithelium, many 
 vibratile epithelial cells, and others as before. Myelin in this 
 sputa was so plentiful as to rival the abundance found in the 
 first case. The bacteria were very much the same as in the 
 other cases, but among them were observed some diplococci, 
 apparently capsular, similar, though smaller, to those shown in 
 / and /'. There were also some slender tufts of leptothrix. I 
 have no record of fungi in these last four cases. 
 
 Other Cases. 
 
 During the month of June I examined other sputa in six 
 different cases of Whooping-cough, and in these I limited thy 
 research to the observation of myelin and fungi. Myelin was 
 found always present in large quantities in severe cases and in the 
 height of the attack, but in more diminished quantities in doubtful 
 cases, and during the beginning and decline of the attack. The 
 larger germinating filaments of fungi were met with in one case 
 only. 
 
 Altogether in fifteen cases of Whooping-cough, nineteen 
 examinations of the sputa were made, and the results may be 
 briefly stated to have been : — 
 
 (i) That myelin is not found to show any relatiori with fever 
 or inflammatory symptoms which may accompany an attack of 
 Whooping-cough, nor to bear any proportion to the quantity of 
 the ellipsoidal cells of epithelium ; but to show evident relation 
 and proportion to the intensity of the nervous excitement. 
 
22 SPUTA OF WHOOPING-COUGH. 
 
 (2) That the bacteria and the baciUii in Whooping-cough 
 sputa are of a most varied character and such as are found in 
 other sputa and in many other substances. 
 
 (3) That the large germinating filaments of fungi have been 
 observed only in a minority of cases — in three cases out of 
 fifteen, and in five specimens of sputa out of the nineteen 
 examined ; they were, therefore, undoubtedly absent in the 
 majority of cases. 
 
 The following sections will be devoted to the general 
 discussion of myelin, bacteria, and fungi in the sputum of 
 Whooping-cough. 
 
 § III. 
 NOTES ON MYELIN. 
 
 Myelin in general. 
 
 DR. Beale, in ^is treatise, " The Microscope in Medicine," 
 thus describes myelin:— "Myelin is a colourless, glistening, 
 semi-fluid substance, prone to form drops, and capable of 
 being drawn out into long threads, which curve and twist into the 
 most peculiar forms. If the observer examines a portion of the 
 white matter of the brain or spinal cord in water, he will recog- 
 nise this substance without difficulty. The masses often exhibit 
 double contours, and not infrequently many lines may be discerned 
 equidistant from one another, but varying much in thickness and 
 in intensity (PI. XXI., Fig. 6a). It is present in the liver, and 
 may be detected in almost all the tissues. In many tissues of the 
 adult, myelin exists in considerable quantity in the outer part of 
 the cells (Fig. 4), and in old age a still larger proportion is present. 
 This substance was first described by Virchow, but Beneke has 
 shown that it may be obtained from all the tissues of the body, 
 and that it exists even in plants. It is soluble in hot alcohol, 
 ether, and turpentine. Cholesterine is a necessary constituent of 
 myelin, and can always be obtained from it. Iodine tinges 
 myelin of a reddish-brown colour. If sulphuric acid be added, a 
 blue or violet colour is induced. This reaction probably depends 
 upon the presence of cholesterine. Beneke showed that myelin 
 gave the reaction characteristic of the biliary acids upon the appli- 
 cation of Pettenkofer's test." * 
 
 * Beale, loc. cit., page 189. 
 
SPUTA OF WHOOPING-COUGH. 23 
 
 The words gocdole, gouttelettes, drops, myelintropfchen, used by 
 the English author and by Virchow and others,* do not seem to 
 me to express so well the physical structure of myelin and its many 
 forms as that more suitable one which Ranvier adopted, viz.— 
 filamentous buttons. Ranvier, in his treatise on Histology, thus 
 expresses himself on myelin in nerve-fibres :— 
 
 " When subjected to the action of water, the axis cylinder, 
 being laid bare, swells until the myelin surrounding it is broken 
 up into filamentous buttons. They may be described as trans- 
 parent threads wound up upon themselves. These threads swell 
 little by little, the outlines becoming less and less distinct till they 
 seem to merge into each other, and n about half-an-hour or an 
 hour the filamentous buttons arise, as little balls of various dimen- 
 sions, showing a very refractive outline, the concentric striae cor- 
 responding somewhat to the original threads. These masses of 
 myelin take the most varied shapes, from that of a cylinder to that 
 of a sphere, the bizarr'e forms in which they present themselves 
 defying all description. Finally, myelin, when isolated, is entirely 
 transformed into spherical or cylindrical masses of different 
 lengths, showing a double contour, presenting a highly refractive 
 border of varying width. This gradual transformation of myelin 
 does not in the least resemble coagulation. On the contrary, all_ 
 the fibres that at first are to be seen absorbing the water seem to 
 swell, to run together, and so produce these rounded masses with 
 the characteristic double outline. 
 
 It is only where the nervous fibres have been divided by sec- 
 tioning or tearing that the myelin breaks away in the additional 
 liquid and takes the various forms that we have already spoken of 
 The natural contour of the fibre remains regular, however, during 
 the working of the preparation, unless, by chance, there is some 
 other lesion from which other threads of myelin may flow, as from 
 the part operated upon."t 
 
 It is true we owe to Virchow the first description arid the very 
 name of myelin. | Nevertheless, myelin had been discovered 
 
 * Jaksch, Manuel de Diagnostic des Malad. Inter., par les Methodes Bacfe- 
 riologiques, etc. Trad. del'Allemand, Paris, 1888, p. 63. 
 tRanvier, Traits Technique d" Histologie. Paris, 1875—82 (ending '89), p. 720. 
 t Virchow, Archiv, 1854—6, p. 562. 
 
24 , SPUTA OF WHOOPING-COUGH. 
 
 before, for these masses, filaments, and drops which escape from 
 the nerve-sheath are described in the middle of this century by 
 Henle in his " General Anatomy." He speaks of them as the 
 globules of medullary substance, which coagulate.* 
 
 Under the word myelin in Selmi's Chemical Dictionary, we 
 find the following remarks : — " Myelin may be obtained by treat- 
 ing the yolk of a boiled egg with alcohol and subsequent evapora- 
 tion, or in like manner from the substance of the brain, crystalline 
 humour, and from other tissues. Beneke has found it in the 
 snail, Helix pomatia, in other lower animals, and in the buds, 
 flowers, and seeds of plants, together with cholesterine. Liebreich 
 thought it to be a combination of protogon, and its products of 
 decomposition (stearic acid, phosphoglyceric acid, and neurin). 
 Neubauer obtained it from oleic acid by simply placing small por- 
 tions of the acid side by side on the slide, and covering them with 
 a drop of liquid ammonia. He likewise obtained it from a mixture 
 of caproic and capric acids and ammonia. Hence we may con- 
 clude that myelin is a special form derived from modifications of 
 fatty substances." t 
 
 Having repeated this experiment of Neubauer with oleic acid 
 and ammonia, I can testify to the production of the various forms 
 of myelin, paler in colour, and with a prevalence of fringed forms. 
 When gentian violet is allowed to run into the preparation, some 
 of the particles are stained to a deep blue, as occurs with sputa ; 
 but the greater number are stained violet or brownish red, these 
 particles having a tendency to become more liquid and run to the 
 edge of the portion under examination. The solution of iodine 
 in potassic iodide, allowed in like manner to flow into a prepara- 
 tion, dissolves all masses of myelin, while it permanently stains 
 those of the sputa. But if, instead of allowing the ammonia to • 
 run by capillary attraction between the slide and the thin cover, a 
 drop of it is allowed to fall on the oleic acid without the cover- 
 glass, the whole becomes a saponaceous mass, wherein no trace of 
 myelin can be seen, it being dense and too opaque. 
 
 * Henle, Trait, di Anat. gen. edit. Levi, Venice, 1845, Vol. II., pp. 144 — 45, 
 PI. IV., Fig. 5, in which are drawn many of the forms taken by myelin when 
 escaping from incisions in the nerve-iibres ; magnified 300 diam. 
 
 + F. Selmi, Encidopedia di Chimica, Turin, 1873, Vol. VII., pp. 883 — 84. 
 
SPUTA OP WHOOPING-COUGH. 25 
 
 Myelin in Whooping-Cough Sputa. 
 
 It has been seen that myelin is not always distributed uni- 
 formly nor to the same extent in certain sputa. If on emptying 
 the sputa from the sputa-cup, striae of a hyaline mucus are noticed 
 adherent to the bottom or to the walls of the vessel, it probably 
 will be found that myelin is more abundant in them, as myelin 
 will render such portions, where it is in larger quantities, consider- 
 ably more viscid than the rest. But if a fresh portion of the sputa 
 is examined in its natural state, after a very short time the parti- 
 cles, filaments, and free granules of myelin, and, above all, the 
 masses, will rise to the surface of the specimen, flowing at times 
 over the lower matter to the edge of the thin cover-glass, following 
 the currents produced in the preparation caused either by greater 
 evaporation going on one side than the other, or by air-bubbles, or 
 by motion imparted to the mass under the hands of the manipu- 
 lator ; so it is clear that myelin is much lighter than the rest of the 
 matter of the sputa-, and being once free rises to the surface. In 
 a conical glass or in a test-tube myelin will very soon rise to the 
 surface, as the fatty globules do in milk, of course with the 
 exception of such particles that are in close adhesion to the epi- 
 thelium cells or firmly embedded in the mass of the mucus. 
 When to a dense mass of mucus some diluent — as, for example, a 
 solution of chloride of sodium — is added, very soon we have a 
 central mass of viscid matter surrounded by the more fluid diluent, 
 into which the lighter particles and granules of myelin will flow, in 
 the same manner as all the lighter and more fluid particles make 
 their way to the edges of preparations. 
 
 Such particles and granules that remain behind are those that 
 are deeply embedded in the mucus. They may be seen princi- 
 pally'in the striae, moving at times along the course of these till 
 stopped by the crossing and closing up of the tiny channels formed 
 in the viscid mass. These particles would easily escape observa- 
 tion in the natural state, but when stained become distinctly 
 visible, especially if gentian violet is used, for the edges of the 
 stria take a more or less blue tinge, and the imprisoned myelin 
 can hardly be overlooked. The granules of myelin adhering to 
 the ellipsoidal cells of epithelium are well shown, too, by staining 
 the mass with gentian violet on account of the colour imparted to 
 
26 SPUTA OF WHOOPING-COUGH. 
 
 such cells. In fresh preparation, especially in the unstained 
 sputum, after some little time, these granules of myelin will leave 
 the cells of epithelium and float off into the surrounding fluid. 
 
 The general appearance of the masses and particles of myelin, 
 in its free state or in sputa, is well described in the passages given 
 from Beale and Ranvier ; but it would be impossible to give a 
 complete idea of all the many forms in which it may be found. 
 
 In the natural state the masses and particles have the colour 
 of oUve oil at the edge, but the central portion seems to be of a 
 light greyish tint, but this changes under the microscope, as the 
 focus is altered by the slow motion. These are shown in Fig. i, 
 a, a, a, a, and c. When the mass is in repose, or is moved without 
 distortion, it is usually marked by concentric striae. But if, as in c, 
 an extremity, d, of a mass of myelin is caught by some foreign 
 body, a current of the containing fluid will push forward the rest 
 as a rounded head, as in e, leaving a narrow neck linking it to the 
 part held firm by the obstruction. The neck becomes thinner and 
 thinner as the progress of the moving portion of the mass con- 
 tinues ; the striae in this portion being distorted more and more, 
 taking at last the appearance of closely packed threads. The 
 neck may be stretched till its length becomes some ten times that 
 shown in the figure, and is proportionately thinner, and at last 
 snaps in two. These broken bands, when stained by gentian 
 violet, are quite distinct ; at times they seem to become solid, as 
 in b, or form new smaller globules, as in b above, on the left. 
 Near the middle of the group shown may be seen a small portion 
 of myelin with two wings, and which in the preparation seemed 
 endowed with a rapid vibratory motion. I'hese threads and fila- 
 ments are, of course, hardly visible when left unstained. 
 
 Besides the so-called masses, many of which are much larger 
 than that shown in c (mag. 400 diam.), there are found aggrega- 
 tions of particles, as in a on the right, of the most varied shapes 
 and sizes. Such united particles rarely coalesce, but take by their 
 mutual pressure a thousand different shapes. 
 
 In the unstained state the granules of myelin are even less 
 visible than the filaments ; in fact, are too minute to be visible. 
 A solution of iodine, picric acid, and fuchsine are a great help, as 
 we shall see in the sixth section ; but gentian violet is of a far 
 
SPUTA OF WHOOPING-COUGH. 27 
 
 greater value, as it gives to the granules and particles of myelin a 
 more or less blue tinge, whilst bacteria and other morphological 
 particles become decidedly violet under its action. This is shown 
 in Fig. I, where b, b, b, b, b should be compared with /and h in the 
 centre and right of the same figure, and then with/ and with the 
 stained bacteria in Fig. 2, especially with s, where the isolated 
 bacteria and the beaded forms are tinted with violet, while the 
 granules of myelin which interpose are stained pale blue. Without 
 this process of staining, the granules of myelin would easily be 
 passed over or mistaken for minute micrococci. 
 
 Myelin in other Sputa. 
 
 It must not be thought that myelin, as described above, is 
 exclusively found in the sputa of Whooping-cough, for it is well 
 known that traces of it are found in all kinds of sputa. 
 
 {a) I have frequently examined the sputa of persons in the 
 best of health, both during the day and immediately after rising in 
 the morning. In such sputa the elUpsoidal cells of epithelium are 
 constantly found covered with myelin, as well as some few parti- 
 cles or free granules of myelin. Nevertheless, I have never found 
 such profusion of masses as in typical cases of Whooping-cough. 
 In all probability, the free granules of myelin in healthy 
 sputa may be held to be simply proportions that have become 
 detached from the epithelium cells, whereas in cases of Whoop- 
 ing-cough the masses are out of all proportion with the 
 number of cells. Again, the proportion of such sputa in 
 healthy subjects is very small. During the whole twenty-four 
 hours the entire quantity would hardly be altogether more than 
 one single expectoration of sputa in Whooping-cough, so that the 
 total amount of myelin expectorated by a person in health is but 
 a very small fraction of that produced in Whooping-cough. 
 
 {b) In acute attacks of bronchitis the ellipsoidal epithelium 
 cells are found covered with myelin and also granular masses of 
 the same, as Bizzozero's * note testifies, but this only in very small 
 quantities as compared with that met with in cases of Whooping- 
 cough. And then it must be remembered that even in cases of a 
 severe character, when the ellipsoidal cells of epithelium are want- 
 
 * Bizzozero, Manuals di Microscopia Clinica, Milan, 1882, p. 154. 
 
28 SPUTA OF WHOOPING-COUGH. 
 
 ing, no myelin whatever is seen. So also in chronic bronchitis 
 myelin diminishes and disappears altogether, as the ellipsoidal 
 epithelium cells diminish or fail to appear. 
 
 (c) In cases of inflammation of the lungs, in which sputa are 
 so frequently examined, it is well known how small the amount of 
 myeUn is, though in such cases the above cells are more than ever 
 abundant. 
 
 {d) In no genuine case of pleurisy has myelin ever been found. 
 The mas's of the sputa may be said to entirely consist of mucus 
 overrun with bacteria, which are, for the greater part, to all 
 appearance, capsular. 
 
 {e) In phthisis, where the sputum is most often examined, 
 myelin is rarely met with, and when found is only present in 
 minute granules, and can only be discovered by the use of gentian 
 violet in fresh expectorations. Only one exception to my expe- 
 rience do I know of- — that mentioned by Dr. Beale *, in which 
 case the sputa must have contained, , according to his report, an 
 amount of myelin approaching that which I have found in 
 Whooping-cough. 
 
 I am therefore inclined to think that, both in health and 
 disease, there may be a continual, though very limited, flow of 
 myelin in the air-passages, as is found in the liver and other organs 
 according to Beale. It is a very different matter when we con- 
 sider the sputa in cases of Whooping-cough. 
 
 In these cases we have no transitory formation (as in the 
 above case spoken of by Beale), but the production of several 
 grammes of myelin within the twenty-four hours, and, what is 
 much more, a daily repetition of it for some months. If we 
 suppose some twelve paroxysms a day, and the amount of sputa 
 at each paroxysm to be about five grammes, there would thus be 
 sixty grammes of sputa expectorated ; and the preparations des- 
 cribed above show that of this amount a very large proportion 
 consists of myelin. What is the cause of this appearance ? 
 
 I will here briefly say what at once came to my mind in answer 
 to this question. First, we have a disease in which an extra- 
 ordinary formation of myelin is produced ; and secondly, a tissue 
 affected which is most rich in myelin, namely, the nervous tissue. 
 * Beale, loc. cit., p. 282, and PI. XL., Fig. 8. 
 
SPUTA OF WHOOPING-COUGH. 29 
 
 The very character of the disease (Whooping-cough) is the intense 
 excitement of this tissue. May it not be supposed that this rapid 
 and intense deterioration or disintegration of this tissue gives the 
 key to the solution of the question ? 
 
 Though Beale does not consider the existence of an alveolar 
 epithelium sufficiently proved, nevertheless, he admits that par- 
 ticles of myelin are found in the air-passages and air-cells of the 
 lungs. Such are shown in i in Fig. i * of the plate, so that this 
 abundance of myelin plight well be thought to be merely the 
 result of physiological action, the increased excitement of the 
 nerves of the part affected bringing about a disintegration of the 
 medullary sheath. 
 
 The Importance of the Myelin in Whooping-Cough. 
 (a) There is no doubt that the far greater part of medical 
 authorities hold Whooping-cough to be due to either a direct or 
 reflex nervous action. 
 
 Cullen, Brouzet, Pinel, and numerous others are of opinion 
 that Whooping-cough is caused by an irritation of the mucous 
 membrane of the gastro-intestinal tract, and Padalme adds to 
 these causes irritation of the lungs and diaphragm. Watt, Alcock, 
 and Laennec consider the disease to be a spasmodic variety of 
 bronchitis ; and Brousseais thinks, in addition, that an increased 
 irritability of the mucous lining of the inflamed bronchial mem- 
 brane is another cause. Trousseau and Richard de Nancy 
 believe it to be but bronchitis grafted on a neurosis.f 
 
 Giuseppe Frank quotes the cases of autopsy made by Hermann 
 Kilian, who in the bodies of fifteen boys, victims of Whooping- 
 cough, found the pneumogastric nerves highly inflamed. | 
 
 G. Pietro Frank places Whooping-cough among the nervous 
 diseases. § 
 
 Copland, in his Dictionary of Practical Medicine, states that he 
 has found in nearly all the bodies of those dead from Whooping- 
 
 *Dr. Beale, loc. Hi., p. 389, PI. XXI., Fig. 6. 
 
 t Barlow, Lancet, 1S86, Vol. I., p. 870. 
 
 X G. Frank, Prax. Med. Univ. Pmcept., Lipsise, 1823, Part II., Vol. II., p. 833. 
 
 § G. P. Frank, Epitome di Medicina Pratica, Chiaverini edition, Naples, 1833, 
 
 Vol. XIII., pp. 153, etc. 
 
30 SPUTA OF WHOOPING-COUGH. 
 
 cough severe inflammation of the cerebral membranes and of the 
 medulla oblongata.* 
 
 Hagenbach, in above-mentioned monograph, mentions many 
 authors of the same opinion. " The seat from which spring the 
 attacks of Whooping-cough (says Biermer) has been placed in the 
 vagus and recurrent nerves, as well as in the phrenic or in the 
 intercostal nerves, in the sympathetic and in the solar plexuses. 
 Webster (London Med. and Phys. Journal, December, i8z2, 
 p. 478) holds " that the actual seat of the complaint may be 
 in the head, and that the affection of the respiratory organs is only 
 to be considered as a secondary effect." Hufeland and others 
 attribute it to excitement of the respiratory nerves. Fried- 
 leben (Archiv. ficr Physiologische, Heilk.^ Volume XII., 1853) 
 thinks it due to a compression of the vagus and recurrent nerves 
 by the swelling of the glands of the bronchise and tracheae. 
 Bouchut thought it to be a complication of catarrh and nerv- 
 ous affections. Henoch (Beitrdge zur Kinderheilk. Neue 
 FolgCy 1868) thinks it to be an affection stimulating in some 
 unexplained manner the nerve-centres of the medulla oblon- 
 gata, producing spasmodic coughing. It is true that Hagenbach, 
 relying on the observations of Letzerich, thought Whooping- 
 cough a mycotic catarrh, but admits the presence of a nerve 
 stimulus given to peripheral ends of the superior nerve of the 
 larynx. \ 
 
 Rilliet and Barthez, in their treatise of Pediatrics, have placed 
 Whooping-cough as a class of disease between the acute exanthe- 
 mata and the neurotic diseases. The following observation of 
 these authors are worthy of great consideration : — " Whooping- 
 cough may show itself at any period of infancy. We have 
 observed it in a new-born child whose mother had suffered from 
 Whooping-cough a month before her confinement. The quintes 
 showed itself in a most violent and distressing manner on the day 
 of birth." I 
 
 * J. Copland, Dictionary of Practical Medicine, Part V. , Art. Whooping- 
 cough, London, 1838. 
 + Gerhardt, loc. cit. , pp. 479 — 80. 
 
 X Rilliet and Barther, TraiU Clinique et Pratique des Malad. des Enfants, 
 Paris, 1853, Vol. II., p. 644. 
 
SPUTA OF WHOOPING-COUGH. 31 
 
 Bouchut has known cases where Whooping-cough developed 
 itself in a child but two days old, but he says nothing of the 
 quintes.* This sudden appearance of the disease distinguishes it 
 at once from such maladies as are contracted from living germs 
 which require for development a period of incubation. We 
 cannot suppose any infection communicated during interuterine 
 life, for such infection would seem impossible in a disease so local 
 as Whooping-cough, and is only known in infections more general 
 in their nature, as in syphilis, acute exanthemata, etc. ; whereas 
 any disturbance of nerve excitability may well be transmitted by 
 the law of sympathy from the nerves of the mother to the homo- 
 logous nerves of the foetus. Henning was of opinion that even 
 the sound of Whooping-cough may cause in another subject a 
 stimulus of the bronchial nerves, f Niemeyer, though he held 
 Whooping to be an infectious cough, speaks even of using the 
 whip in this malady. J 
 
 Delaberge, Monneret, and Fleury, in their compendium of 
 Practical Medicine, mentions a case, referred to by Gendrin, in 
 which a persistent cough, with all the characteristics of Whooping- 
 cough, was observed by Dupuytren and Husson to result from an 
 operation for opening the parotid gland. § 
 
 " Unless we reject the evidence of well-established cases, it is 
 impossible," these authors add, " to refuse to look upon Whooping- 
 cough as a nervous affection, more especially if we consider— ist, 
 That the respiratory parts present no alteration, or if any are 
 present they are of so varying a character as not to be considered 
 a cause of the disease ; 2nd, That the course of the symptoms is 
 evidently remittent, but with such an absence of fever, when any 
 complication does not exist which is not observed in inflammations 
 of a general or specific character; 3rd, The sudden cessation and 
 return of the spasms of coughing, brought about by emotion or by 
 change of place, are phenomena that point to nerve excitement, 
 * Gerhardt, loc. cit., pp. 475—76. 
 t Ibidem, p. 480. 
 t Niemeyer, Patologia e Ter. spec. int. Versione Cantani, Milano, 1863, 
 
 Vol. I., p. 38 (in a note). 
 
 § Delaberge, Monneret, and Fleury, Comfendzo di Med. Pralica, Versione 
 
 it. Firenze, 1,851, Vol. VI., p. 3I4- 
 
32 SPUTA dF WHOOPING-COUGH. 
 
 and not to inflammations that require time to run out their course ; 
 4th (and lastly), That rapid regaining of perfect health, and the 
 healthy condition of all functions in mild cases, the resistance of 
 the malady to treatment, and the inefficacy of the usual remedies 
 prescribed for inflammatory diseases, the success of narcotics and 
 antispasmodics, are all circumstances met with in nerve affections, 
 but not in diseases accompanied by inflammation." * 
 
 GrisoUe repeats the same arguments for including the Whoop- 
 ing-cough in the class of neurotic diseases, t 
 
 Jaccoud also admits the specific excitement of the nerves in 
 Whooping-cough. . It is proved beyond doubt by Rosenthal's 
 researches that a centripetal stimulus given to the superior laryn- 
 geal nerve determines a relaxation of the diaphragm, the closing 
 of the glottis, and a convulsive expiration. That is, the pheno- 
 mena that distinguishes a convulsive cough from others altogether 
 catarrhal, are explained by an irritation of the superior branch of 
 the vagus nerve, produced by a reflex action on the medulla oblongata, 
 thus arresting inspiration and causing a spasmodic expiratory action 
 of the glottis. I 
 
 West believes Whooping-cough to be a bronchial and nervous 
 affection. He mentions the fact that the pneumogastric nerves 
 have been found on examination to be reddened, swollen, and 
 softened, but only in exceptional cases. According to him, Albers 
 of Bonn in forty-seven autopsies found in three cases only the 
 vagus reddened on the right side, and in one case on the left, which 
 might have been merely the result of position. Only in one case 
 out of twenty-four, he found the nerve on both sides of a higher 
 colour than in the normal state. Hence, he concluded that such 
 changes were simply accidental or due to alteration after death. 
 He does not make any allusion to the superior laryngeal nerve. § 
 
 Certainly, the sudden beginning and ending of the spasms, 
 which he describes, plainly point to a nerve affection rather than 
 to infection by living germs. || But not to prolong this matter, I 
 refer the reader to the respective authors. 
 
 * Ibidem, p. 336. 
 
 t GrisoUe, Tratt. elem. eprat. di Pat. int. vers. Del Corso, Livorno,'i853, P- 877- 
 
 t Jaccoud, Tratt. di Pat. int. vers. Borrelli, Napoli, 1872, Vol.^I., p. 751. 
 
 § West, Lezioni suite malattie dell' infanzia efaruiullezza, vers. Blasi, 
 
 Milano, 1869, pp. 498—99. || Ibidem, pp. 476 and 493. 
 
SPUTA OF WHOOPING-COUGH. 33 
 
 In the first paragraph 1 mentioned Michael's and Sonnen- 
 berger's opinions as well as that of others, who hold that Whoop- 
 ing-cough is but a reflex nerve affection. 
 
 In a treatise on the pathology and treatment of Whooping- 
 cough during last May, Genser, without entering into any new or 
 microscopic researches, combats the practice of nasal insufflation 
 recommended by Michael, and advocating in their place the use 
 of antipyrin, not as a parasiticide, but on account of its sedative 
 action on the nerves, reducing or suppressing reflex action in them. 
 This shows how much he considers nerve aff"ection has part in the 
 disease.* 
 
 Cohnheim and Habershon likewise consider Whooping-cough 
 to be of the nature of a nerve derangement. Habershon, more- 
 over, thinks it proceeds from some derangement in the function 
 of the pneumogastric nerve, f 
 
 The hypothesis of the breaking up of the medullary sheath, 
 replete with myelin, of the nerve-fibres in the air-passages, includes 
 the supposition of some morbid excitement of these nerves. 
 Nerve-fibres, with the exception of the fibres of Remak, are 
 invested with a medullary sheath, forming a layer of considerable 
 relative thickness between Schwann's sheath and the axis-cylinder, 
 being interrupted by the constrictions of Schwann's sheath at 
 regular intervals ; the fibres of Remak are readily distinguished 
 among the meduUated nerves by the absence of the medullary 
 sheath and by their anastomosing plexuses. % 
 
 Here we are confronted with a difficulty of explaining the 
 passage of the myelin, escaping through such alterations from the. 
 nerve-fibres into the interior of the air-passages. It would appear 
 that myelin must be included amongst the colloid rather than the 
 , crystalloid substances ; its power of passing through animal mem- 
 branes would then be very little. 
 
 But such a difficulty would militate chiefly against that portion 
 
 * Genser, Zeitschr. f. Ther., No. 9, 1888 ; Riforma Medica, July, 1888 ; 
 and Morgagni, Part II. , No. 39. 
 
 tCohnheim, Lez. di pat. gen.. Vol. II., Naples, 1882, pp. 149, 150. 
 Habershon, Lez. sulla pat. del n. pneumogastrico, Milan, 1879, pp. 19, 20, 
 
 and p. 30. 
 
 tRanvier, loc. cit, p. 772, etc. 
 
34 SPUTA OF WHOOPING-COUGH. 
 
 Of the myelin which is supposed to be formed at the expense of 
 the nerve-fibres which surround the alveoH, or the termmations of 
 the ultimate bronchials; where the existence of a mucous mem- 
 brane, properly so.called, cannot be demonstrated on the mternal 
 surface of the membrana anista. It is not the same with the 
 whole of the remaining bronchial tree, the trachea and the larynx, 
 which are invested with a mucous membrane very rich in nerve- 
 fibres. 
 
 In the epiglottis, according to an example drawn by Beale, the 
 network of nerves lies on the surface of the mucous membrane 
 immediately underneath the epithehum.* 
 
 In the mucous membrane of the larynx and trachea it is less 
 superficial, but in the bronchi it again comes to the surface with a 
 thinning of the mucous membrane.f 
 
 Hitherto we have only expressed a doubt as to the origin of 
 the myelin in the sputum of Whooping-cough, and we refrain 
 from entering into further disquisition on the subject, for which 
 there would be no occasion, except the fact stated by us of the 
 extraordinary abundance of myelin in such sputum, was confirmed 
 by further and more authoritative observations. 
 
 (b) The abundance of the myelin once proved will also give 
 us the explanation of the singular character which the sputum of 
 Whooping-cough always presents. Its viscosity, which presents an 
 impediment to expulsion, does not depend on the condensation of 
 the mucus, nor has it anything in common with the thick and 
 tenacious mucus which is commonly met with. The sputum of 
 Whooping-cough is, on the contrary, fluid, thin, and transparent. 
 
 * Beale, loc. cit., p. 384, PI. LX., Fig. 2. The same in the Lectures on 
 the Structure of Simple Tissues, etc., Borrelli, ed., Naples, 1865-, p. 284, 
 PI. XV., Fig. 95. 
 
 tKoUiker, EUm. d'histolog. hum., Vax'is, 1871, pp. 510 — 529. Sappey, 
 Traite dAnat. Descrip., Paris, 1889, torn. IV., p. 439. Stohr, 1st. dHstolog., 
 etc., Naples, 1887, p. 164. Concerning the degeneration and reproduction of 
 nerve-fibres, the travelling of the myelin particles, and the very slow repro- 
 duction of the medullary sheath, see, amongst other authors, Tizzoni, Sulla 
 paiolog. deltessuto nervosa, etc., Arch. p. le Sc. Mediche, Turin, 1879, Vol. III., 
 No. I ; Alonzo, Sulla degeneraz. dellef. nervose, etc.. Arch, citato. Vol. XIII., 
 1889, p. 229, etc. 
 
SPUTA OF WHOOPING-COUGH. 35 
 
 Now, these properties, combined with its viscosity, are perfectly 
 understood if they relate to the myelin. 
 
 Further, the presence of a notable quantity of myelin will 
 furnish a critical diagnosis in certain cases. 
 
 Rilliet and Barthez have established the great difficulty of a 
 differential diagnosis in grave cases of pulmonary complications 
 which are evolved in the early stages of the disease, and are pro- 
 tracted in the later stages, resembling capillary bronchitis with fits 
 of coughing (with quinies). The differential diagnosis between 
 Whooping-cough and tuberculosis of the bronchial ganglia is yet 
 more difficult.* 
 
 Hagenbach, in order to make the diagnosis more certain in 
 doubtful cases, considers it indispensable to be present during a 
 paroxysm, and if required to provoke it with some stimulus, 
 Whooping-cough being liable to be confounded with capillary 
 bronchitis, pulmonary phthisis, tuberculosis of the bronchial 
 glands, hysterical cough, and relaxed uvula.t 
 
 Now, in all these cases or others similar, the microscopical 
 examination of the sputum, whenever there is a great abundance 
 of myelin, will easily solve the question. 
 
 Researches on Expired Air. 
 
 We shall see in the sixth section that the minutest particles of 
 myelin — so minute that they can scarcely be observed unless 
 stained — are animated by a lively molecular motion. Its extreme 
 tenuity and motobility in liquid currents has already' been noticed. 
 It cannot be thought impossible, nor even improbable, that these 
 minute particles of myelin may be expelled with the expired air, 
 especially during the attacks of coughing, and be diffused in and 
 remain suspended in the surrounding atmosphere, for such parti- 
 cles are less, both in weight and volume, than the small bacteria 
 found in the air. 
 
 For want of better apparatus, I have made use of large plates 
 of water, to which has been added a small quantity of phenol, and 
 
 * Rilliet and Barthez, loc. cit., p. 639. 
 tGerhardt, loc. cit., pp. 495—496. 
 
36 SPUTA OF WHOOPING-COUGH. 
 
 kept them some ten days in the rooms of patients. But on exa- 
 mination of the flocculent deposit in this water, which was poured 
 into a conical glass, I have not found either particles or granules 
 of myehn. Many living amoeba were found ; also spores (of 
 which the greater part was very similar to the spores of the larger 
 fungi observed in the sputa), as well as two forms of bacteria. 
 One of these was dumb-bell shaped, and similar to the types ^and 
 e in the plate. Fig. 2, and most likely the same as that found by 
 Poulet ; the other showing spotted granules very much after the 
 manner of the supposed bacillus of typhus fever. I need not 
 give a full Ust of other substances observed, such as pollen and 
 starch-grains, etc. In the doubt that the particles of myehn 
 might have remained floating on the surface of the water, this, too, 
 was examined without any discovery of myelin. 
 
 But this examination was too imperfect to enable me to draw 
 any definite conclusion. I leave others, better equipped with 
 suitable apparatus, to settle this point, and to enter into the ques- 
 tion whether such a diffusion of myelin in expired air, if proved, 
 might not be a means of spreading the disease. 
 
 To ascertain, moreover, whether vapours could diffuse the 
 granules of myelin, some sputa were diluted with distilled water 
 and boiled in a test-tube for some two or three minutes. A glass 
 slip was applied to the mouth of the tube and held there by a 
 spring. The examination of the dewy film in the glass slip did 
 not reveal any trace of myehn, but I observed some minute 
 globules, oleaginous to all appearance, which seemed to tend to 
 unite in larger globules, and were not at all stained with gentian 
 violet. I cannot say that these were formed in any way by the 
 boiling of the myelin, but of this I am sure, that the quantity of 
 the myelin in the boiled liquid was diminished. 
 
 It will not be out of place here to mention an incident which 
 took place during one of these examinations. Having inadver- 
 tently taken some ordinary cistern-water in place of distilled 
 water, I found in one of the preparations a fungus as shown in its 
 natural state in Fig. 3a, magnified 400 diameters. I have had 
 occasion to speak of this fungus in another communication to the 
 Society in reference to certain diplococci, similar to the gonococci 
 
SPUTA OF WHOOPING-COUGH. 37 
 
 of Neisser, and observed by me in a case of cancer of the bladder.* 
 The spores of this fungus, being light, float on the surface of 
 liquids and are taken up by pipettes, or glass rods, and deposited 
 on test-tubes or on glass slips for examination. Very likely the 
 warmth from the boiling favoured the germination of the fungus, 
 as shown in the figure, for in all the many preparations examined 
 I have only found this fungus in germination once before, in a 
 dried portion of sputa of a consumptive patient. 
 
 The casual germination of the fungus in this sputum, diluted 
 and boiled as it was, offers by analogy an explanation of the ger- 
 mmation of the fungi, k and n in Fig. 3, the sole difference being 
 that the fungus a was deposited in the sputum after expectoration, 
 and those in k and n received internally. 
 
 §4. 
 OBSERVATIONS «& CONSIDERATIONS ON THE BACTERIA. 
 
 IN the second section I have briefly described the principal 
 forms of bacteria and bacilli observed in the sputa in 
 Whooping-cough. It will now be well to take a more sys- 
 tematic view of their forms, and compare them with others found 
 in different sputa and other substances, not with the object of 
 raising the question of their pathogenic power in engendering 
 disease (which I do not uphold), but to show the value of their 
 chnical importance in the diagnosis of the disease. 
 
 Bacteria and Bacilli found in Whooping-Cough. 
 All the forms of bacteria represented at a, b, c, c, d, Fig. 2, 
 with exception of c , are very frequently met with, and can hardly 
 be said to differ much from Bacterium termo. They are found in 
 all kinds of sputa, and in substances of widely different character ; 
 and it should be noticed that they do not differ from the forms e 
 and 75 which are found covering and infesting the buccal epitheli- 
 um cells and the salivary corpuscles, but do not in any way attack 
 the vibratile epithelium cells, nor the pavement cells of the air 
 passages. 
 
 * Sopra un diplococco analogo al gonococco del Neisser, riiwenuto nella orina 
 in un caso di carcinoma della vescica. See Tom. XLIII. , Atii R. Accad. Med. 
 Chir., 1889. The discovery of such curved forms in diplococci, even in the 
 sputa of Whooping-cough, is fatal to the supposition of their being a specific 
 characteristic of any disease. 
 
38 SPUTA OF WHOOPING-COUGH. 
 
 This was observed before by Barlow, as stated in the first 
 section of the memoir,* who was of the opinion that the epithe- 
 lium cells infested by these bacteria were only those of the larynx 
 and pharynx ; but comparing my observations, on the sputa of 
 Whooping-cough with those on other sputa, healthy or otherwise, 
 I cannot in the least agree with the illustrious doctor of Man- 
 chester. It is quite sufficient to compare these pavement cells, 
 large and thick, with those of the saliva, to assure oneself that they 
 proceed from the buccal cavity, and not from either the pharynx or 
 larynx. Moreover, we must remember that if the epithelium cells 
 of the larynx or pharynx become detached by a supposed specific 
 pathogenic action of these bacteria, they could not in any possible 
 way undergo any regeneration to allow them fully to develop, 
 especially considering that the mucous membrane would be in a high 
 state of irritation. How then can we account for the constant 
 presence in the sputa of such fully developed epithelium cells, 
 and which are always infested by swarms of these bacteria ? 
 
 This fact is to me a most convincing proof of the buccal 
 origin of these bacteria. For if they originated in the sputa, they 
 would doubtlessly be found more frequently among the epithelium 
 cells of the air passages than among those of the mouth, or in the 
 salivary corpuscles, or at the very least as often among the first as 
 among the latter. The former could hardly be attacked by the 
 bacteria in the short period elapsing between the expectoration 
 and the immediate examination, and such immediate examination 
 of the sputa shows that while epithelium cells of the mouth 
 swarm with such bacteria, the epithelium cells of the air passages 
 are entirely free from them. 
 
 We must conclude, 'therefore, that the germs of such bacteria 
 found free in the sputa come from the mouth, having been 
 inspired with the air, and lodge within the entrance of the air 
 passages, after the manner in which Dante describes the fate of 
 the souls of suicides: — 
 
 " Into the wood it falls, at no fixed spot ; 
 
 But where blind fortune haply shoots its bolt, 
 There like a grain of corn it germinates." 
 
 — Inferno, XIII., D. Johnson's trans., 1867. 
 * See page 2. 
 
SPUTA OF WHOOPING-COUGH. 39 
 
 This simile, though it may appear to be out of place in a 
 scientific treatise, is an apt one, not only in regard to the bacteria 
 a, b, c, d, but to all other bacteria or bacilli found in the sputa ; 
 .for there are none, we may safely say, that are not often found at 
 the same time in the epithelium cells of the mouth, or in the 
 saliva, and the germs of which may not have been introduced 
 into the air passages by the currents of the air as it is inhaled. 
 
 This view is not only in complete accordance with the 
 observations of Barlow, but with those also of Jansen and 
 Tschamer, and with the deductions which Cornil and Babes drew 
 from the observations of Biirger, as was noted in the first section. 
 
 As before stated, it is not my intention to enter into the 
 pathology of the subject, but I should like to point to one fact, 
 which I believe has not been referred to before by any one, and it 
 is this : — (it seems to me) that there is evidently a great dispropor- 
 tion in the power of producing disease attributed by many to 
 bacteria on the one part, and their extreme smallness on the other. 
 
 The bacteria a, b, c, and d are of medium size as compared 
 with other bacteria ; now if we take a single grain of wheat, 
 weighing some five centi-grammes, we shall see that it is about half 
 as long again as one of these bacteria, on the scale of the figure ; 
 twice as broad and twice as thick, and would be in volume equal 
 to that of six bacteria, and in weight some eight times that of a 
 single •bacterium. A grain of wheat if put into water sinks at once 
 to the bottom, whilst the bacterium will float on the surface. Now 
 the bacteria in the figure are magnified to 2,500 diameters, or 
 6,250,000 times superficially, or 15,625,000,000 in volume. So 
 we must conclude that this number of bacteria would equal some 
 eighth part of the five centigrammes, or the weight of one grain 
 of wheat. In other words, to make up five centigrammes of 
 bacteria, we must have 15,625,000,000 bacteria multiplied by 
 eight; that is to say, 125,000,000,000. Hence in one centi- 
 gramme we may say there must be some one hundred and twenty- 
 five billions of bacteria. These calculations will not appear to 
 be exaggerated when compared with those of Naegeli, which, as 
 is well known, are some twelve times greater. 
 
 How far such extreme smallness can be reconciled with the 
 power of producing disease attributed to these bacteria must be 
 
40 SPUTA OF WHOOPING-COUGH. 
 
 left to pathologists to decide. We hasten to reconsider the clinical 
 and semiological questions. 
 
 Enough was said about the bacteria V and d in the clinical 
 notes of the first case. In c, above, and in d are seen the dumb- 
 bell like bacteria in series ; while in c, below, are seen the bacte- 
 ria, cylindrical in shape, somewhat swollen in the middle, and 
 arranged in shorter and more sinuous threads. Now, it seems to 
 me that from this last form may be derived the other forms, c d, 
 which are magnified 860 diameters. 
 
 I have occasionally come across this chain-like series, shown 
 in c c, in the sputa of consumptive patients ; it is stained with 
 gentian violet. The individual articulations or bacteria differ but 
 little from those shown in c, below, but they are somewhat larger 
 (in the figure they are shown too long), inclosed in a sheath with 
 a well defined outline, and at the points of contact they seem to 
 coalesce, for no septa, but only restrictions — such as are seen also 
 in Fig./' — are discernible. 
 
 Of the bacteria e, f, I, the forms e and / are nearly identical 
 with those shown in b and d\ but are met with, sometimes in large 
 numbers, enclosed in a sheath, whether they are within or without 
 the epithelium cells. They thus have a distinctive feature in this 
 sheath, real or apparent, in common with the bacteria of the types 
 shown in /', /, /', 0, and / '. The sheath seems to be better defined 
 when the specimen is stained with methyl violet, and still better 
 when the medium is less refractive. I have been able, even in 
 water, to detect this semblance to a containing sheath; but glycerine, 
 when it once permeates the preparation, renders the sheath at first 
 almost invisible, and after some hours totally so. But in air it is quite 
 the reverse. In those parts which, by evaporation of the medium, 
 are left dry on the slide, the sheath becomes at first quite distinct, 
 though eventually, when quite dry, both the sheath and the bac- 
 teria become a confused mass. 
 
 Many bacteria, morphologically identical and with the same 
 apparent sheath, are found in the saliva and in the accompanying 
 buccal epithelium cells. I have met with them in all kinds of 
 sputa, and believe them modified forms of the same kind. So in 
 / and /', on the right, there is seemingly the beginning of a union ■ 
 of the extremities, and a tendency to take the form of e and /* 
 
 * See Fig. 33, p. 76, in the previously cited manual by Jaksch. 
 
SPUTA OF WHOOPING-COUGH. 41 
 
 I shall have to refer to other bacteria with a similar apparent 
 sheath. 
 
 The bacteria, g, p, and/', curved and coupled together, closely 
 resemble the so-called Gomcocci of Neisser, supposed to be the 
 specific characteristic of virulent Blennorhoea, and are found, though 
 m small numbers, in saliva and occasionally in the sputa of Whoop- 
 mg-cough. In Fig. / we have a similar but larger form, inter- 
 mediate between the curved forms seen in g and/. I fail to see 
 why these forms, though varying in size, should be held to be 
 distinct species. 
 
 I must draw attention to the curved forms shown in /. They 
 are linked in a chain-like thread, together with bacteria of about 
 the same size, but straight, and, in all but their size, identical with 
 those shown in b, d, and / Now, it seems quite inconceivable 
 that such a chain or beaded thread of bacteria could be made up 
 of individuals of totally different species ; in other words, that a 
 complex organism— a truly Horatian monster, as we should have 
 m the forms/,/, could possibly be the outcome of the fortuitous 
 grouping of organisms of quite different species. We must, I 
 think, come to the conclusion that thfe curved and the straight 
 forms in p belong to the same species. That the straight forms 
 are identical with those in b, d, and /, is established by the fact 
 that they both are found living on the buccal epithelium cells and 
 have the same dumb-bell shape, and take the same sheathed forms, 
 which the curved forms do not. This shows, on one side, that the 
 semblance of a sheath does not of itself constitute a specific differ- 
 ence ; and on the other, that all the so-far mentioned forms, from 
 a to /, in all probability, are but modifications or stages of deve- 
 lopment of one and the same species. 
 
 Fig. / ' shows two bacteria crosswise enclosed in a common 
 transparent capsule ; but they are found oftentimes singly, or three 
 or four together so enclosed. The single or isolated forms cannot 
 be distinguished from those shown in /, above, where these are seen 
 more or less foreshortened, for then the transparent or semi-trans- 
 parent division is invisible until the bacteria is changed in position. 
 Again, the bacteria shown in /' may take the appearance of diplo- 
 cocci, after the style of /' and . /', with disjointed members, 
 whenever they are seen, not sideways, as in the figure, but end- 
 
42 SPUTA OF WHOOPING-COUGH. 
 
 ways, for their club-headed extremities have the same apparent 
 shape as the rounded diplococci. 
 
 The bacteria of the type shown in / are sometimes met with 
 in quite large agglomerations, mingled with other very minute 
 bacteria of the same shape. This we saw in the second section. 
 These are very numerous in sputa in phthisis, inflammation of the 
 lungs, and in other sputa. With methyl violet they take some- 
 times a dense blue, sometimes a violet tint, both tints showing at 
 times in the same preparation. I shall have occasion later to 
 speak of some probable cause of the appearance of the sheathed 
 or capsular forms. 
 
 The form presented in h was, in linear dimensions, almost 
 double the size of the so-called pneumococci, as will be seen if 
 my drawing be compared with Fig. 13 of PI. XV. in the quoted 
 work of Cornil and Babes, under a, b, and k ; bearing in mind 
 that the drawings of these authors are on a scale of something less, 
 viz. : 2,000 diameters. In my drawing a distinct outhne seems to 
 show the extent of the capsule or sheath. 
 
 I was sorely perplexed as to the nature of this body till, in a 
 case of pleurisy, I met with a large number of similar bodies of 
 about the same size as those I had found in Whooping-cough, with 
 this difference, that those in the case of pleurisy were of various 
 shapes, some being elongated and constricted in the middle, others 
 ellipsoidal, rounded, or altogether irregular in outline. I was soon 
 convinced that such bodies were but small pus or mucus corpus- 
 cles, having a nucleus highly stained, the rest pale blue by osmosis. 
 I had used as a dye a fresh preparation of methyl violet, and con- 
 cluded that the body h, stained with gentian violet, was of the 
 same nature, and had taken by mere accident the appearance of a 
 diplococcus with a sheath, compressed in one direction and 
 lengthened out in another by the threads of the mucus in which 
 it was held. 
 
 The beaded forms of bacteria / and k have nothing to distin- 
 guish them from those found in urine, blood, and many other 
 substances undergoing decomposition. In the form / germination 
 seems to be carried on at one end only, and the larger members, 
 which may be taken to be older bacteria, are more deeply stained. 
 In the form k increase seems to proceed at either end, but no 
 
SPUTA OP WHOOPING-COUGH. 43 
 
 specific difference can be seen between the two types. It is easily 
 conjectured that these beaded forms may develop into the chain- 
 like threads in c and d. 
 
 From my observations of these bacteria, or rather bacilli, of 
 the beaded forms, and the difference of colour imparted in stain- 
 ing to the young and to the older members, I am led to think that 
 the germinal matter predominates in the younger ones, and this is 
 less readily acted upon by aniline dyes ; whereas the older mem- 
 bers become invested with some covering which takes up the stain 
 more easily, as we often see is the case in frustules and filaments 
 of vegetable origin. For with a parity of development I find the 
 contrary takes place : the smaller bacteria are more readily stained 
 than the larger forms. 
 
 For the sake of brevity and clearness I will defer till later the 
 description of the Figs, m and n. 
 
 In Fig. o we have one of the curved forms of bacteria enclos- 
 ing a pale-blue substance. It greatly resembles the curved forms 
 g and /, and is not very unlike those contained within the bacillus 
 n on the lower left side of the figure. I have not met with many 
 specimens of this type, and of the type shown in o above still 
 fewer have come under my notice. This last is a true diplococcus 
 enclosed in a capsule, and its nature is unknown to me, for the 
 internal matter as well as the rest is stained. 
 
 The bacteria q is rarely seen in sputa of Whooping-cough, but 
 is more frequently found in other sputa. It has no specific 
 importance, being, as we shall see, but an undeveloped Leptothrix. 
 
 The form r is of very rare occurrence, but from both these we 
 can see that the longer forms of the bacteria, single or linked, as 
 in q and r, do not differ substantially from the more rounded 
 forms, as shown in / and y. 
 
 At first sight, the beaded form s seemed to me to bear great 
 resemblance to the bacillus of Koch, Bacillus tuberculosis. They 
 are found in many various kinds of sputa. These beaded forms 
 may be called in truth bacilli, for when magnified to some 400 or 
 500 diameters, they appear to be one entire, unbroken thread, as 
 also, under the same power does the bacillus of Koch. Under higher 
 powers, both one and the other show well their moniliform struc- 
 ture. This is well seen if my drawing in the Plate (2,500 diam.) 
 
44 SPUTA OF WHOOPING-COUGH, 
 
 is compared with Figs. 12, 13, and 14 of Plate XXII. of Cornil 
 and Babes, bearing in mind that these last are only drawn to 1000 
 diameters. Such beaded forms did not stand the test with dilute 
 nitric acid (one part acid to three of water), and alcohol as we saw 
 in the second section. I shall have to refer to this again in 
 another place. 
 
 These beaded forms and the bacilli of the same type, are 
 frequently observed in saliva, and seem to predominate both in 
 saliva and sputa around or within the buccal epithelium cells, as I 
 observed in the seventh case of Whooping-cough. I have good 
 reason to think that these beaded forms are substantially the same 
 as the chain-like threads in u, u. 
 
 It will appear from this short description, firstly, that all 
 bacteria found in Whooping-cough sputa can be reduced to a very 
 few species, identical with those found normally in the mouth and 
 perhaps also in the nose ; secondly, that until experiments in 
 culture and inoculation, invariably and for a long period of years, 
 give ,us exact and conclusive results to the contrary, we are right 
 in maintaining that none of these forms are specific causes of 
 disease, and that Whooping-cough is not rendered contagious by 
 bacteria.* 
 
 Bacteria and Bacilli found in Healthy Sputa. 
 In healthy sputa, collected in the manner indicated in the 
 foregoing section, the following bacteria are found : — the dumb-bell 
 forms, b, d, e, and f the beaded forms, i and k, and those 
 shown in g and /, I have observed, tbo, in the sputa of healthy 
 persons, colonies and numerous samples of the straight forms of 
 bacteria, /,/. Even in the sputa examined at once, these bacteria 
 were found in greatest numbers in such portions where there was 
 
 * This second conclusion is entirely supported by Mackenzie, of Edin- 
 burgh, in his monograph on sputa in general, who, by the way, makes no 
 mention of myelin.— G. Hunter Mackenzie, Le crachat dans ses rapports 
 avec le diagnost. ; etc., Paris, 1888, page 21. Such conclusion is thus spoken 
 of by Sternberg:—" Dr. Vicentini very properly arrives at the conclusion that 
 no one of the bacteria observed by himself, or by those who have preceded 
 him in examining the sputa in cases of Whooping-cough, has been demon- 
 strated to bear an etiological relation to this &\s,^?&e^"— Proceedings of the 
 American Microscopical Society, July, 1893, P- '5^. [F. V.] 
 
SPUTA OF WHOOPING-COUGH. 45 
 
 a mixture of the saliva ; these portions were marked by the 
 abundance of buccal epithelium cells, salivary corpuscles, and 
 spirilla, elsewhere scarce. Later, during the day after, and the 
 following days, this type of bacterium became more numerous and 
 the colonies more abundant. Both in the fresh and stale sputa 
 the disposition in groups of two, three, and four together, sur- 
 rounded apparently by a capsule, was frequently observed, and 
 rendered very distinct by staining with methyl violet. 
 
 _ The beaded forms, s, were observed in sputa immediately after 
 expectoration, at times arranged side by side, sometimes inter- 
 lacing, oftentimes longer than the specimens shown in the figure. 
 Sometimes numerous examples were found in which the threads of 
 the same minute articulations as form the type s run parallel. 
 In the stale sputa, on the third to the fifth day, many of these 
 threads took the appearance of long ribbons, by the formation of 
 some connecting matter not only between the threads, but on all 
 sides. This matter takes a pale violet colour, and evidently is 
 connected with the production of the young forms of bacilli, after 
 the style shown on the left and lower side of n. The limited space 
 of this memoir will not permit of my giving an illustration, but the 
 type shown in l>', Fig. 2, will give a very fair representation of it. 
 
 To return to the simple chain-like forms, these oftentimes so 
 increase in length, and mat themselves together, that they present 
 much more intricate forms than those shown in u, u, especially in 
 stale specimens of sputa ; the drawing was made fromi a fresh 
 specimen, and magnified to 600 diameters. Such ' interwoven 
 masses are found not only in whooping-cough, as we saw in the 
 second section, but also in the sputa of phthisis, in cases of 
 chronic laryngitis, and in inflammation of the lungs, especially a 
 day or so after expectoration, and in warm weather. 
 
 I beheve that these masses, of the type u, u, spring from the 
 beaded forms s, for several times I have come across specimens 
 just at the period of transition. Want of space prevents me from 
 giving a drawing of these forms; so, too, many specimens of 
 ribbon-like forms are not given for the same reason. These forms 
 lead me to think that the bacilli of type n, on the right side of the 
 plate, and below, are derived, as I have said, from the single 
 articulations of the type u, u. There still remains the doubt 
 
46 SPUTA OP WHOOPING-COUGH. 
 
 whether the curved bacteria as at o in the lower part of the plate, 
 coupled and surrounded by a cementing medium, can give origin 
 to identical bacilli. 
 
 Again, in healthy sputa I have found short chains of ellipsoidal 
 bacteria, v, among such masses as are shown va.u,u; such chains 
 should be looked upon as the origin of the bacilli of type n, as 
 seen on the left of the drawing. These bacteria contain, in fact, 
 a close succession of similar bacilli, and a paler surrounding 
 medium ; it should be observed that the chain, v, is drawn on a 
 scale of 600 diameters, whereas the bacillus, n, is magnified to 
 2,500 diameters. Such transition is proved by the fact that I 
 have found specimens which clearly showed the formation of the 
 cementing medium among the individual bacteria, and ellipsoidal 
 articulations when the sputum was again examined the following 
 day. Figure b' will give an idea of such' specimens, though taken 
 from a bacillus of different character, viz., in a case of inflamma- 
 tion of the lungs. 
 
 The isolated bacilli, v\ v', are distinctly the same as those of 
 the type q, taking into account the difference of scale under which 
 they are drawn. In z are shown many of these bacilli in long 
 lines (2,500 diam.), tending to form a chain-like thread ; these 
 appeared in the same sputa on the second day. They gradually 
 increased in size, and by the fifth day, under a temperature of 
 28°C., took the form of distinct threads or tufts of Leptothrix ; 
 thus we must take the forms q, »', and z, to be but stages of one 
 and the same bacillus, viz., Leptothrix. 
 
 The beaded forms, x, and the rounded bacteria of the type y, 
 were also found in healthy sputa, and it is evident that the 
 rounded members of such beaded forms are the same as the cocci 
 in group y. The upper drawing at x deserves special attention, 
 for it shows the identity of the oblong and rounded members in 
 the specimen. We have here, in one and the same chain, an 
 oblong member with three rounded bodies at each extremity, in 
 little or nothing differing from those in Figs, q and z. This, as we 
 have argued before, proves them to be identical. The figure x' 
 proves the same thing, showing as it does two of the rounded 
 forms, and three oblong ones at each extremity. The lower 
 specimen in Fig. x shows identity with the bacteria in Fig. r, by 
 
SPUTA OF WHOOPING-COUGH. 47 
 
 the similarity of the middle {"ortion. All these forms, therefore, 
 q, r, V ', X, X ', y, and z, belong to one and the same species. We 
 shall see that in like manner the bacilli m, n, n', and therefore 
 those of type u, u, and the beaded forms s, belong to the same 
 species. 
 
 From this we must conclude that taking bacteria as the sole 
 guide, the sputum of Whooping-cough does not differ from the 
 sputum in health. In both cases the micro-organisms proceed 
 from the many that habitually thrive in the mouth, and most 
 probably in the nose as well. This conclusion is precisely that 
 which Tschamer arrived at in 1870 from his first observations on 
 Whooping-cough, as I stated in Section I. 
 
 Bacteria and Bacilli found in Nasal Mucus. 
 
 Leaving for the present my observations on nasal mucus in 
 Whooping-cough, I will make a few remarks on the microbes that 
 infest the mucus in healthy subjects, for the germs of the bacteria 
 found in the air-passages partially come from the mouth and 
 partially, as I have said, from the nose. 
 
 The secretion from the nose in health consists principally of 
 hyaline mucus, more or less dense, containing a number of granu- 
 lated mucus corpuscles, which are endowed with a lively dancing 
 motion as soon as they are subjected to any osmotic action. This 
 motion is better observed after lightly staining, as their granules are 
 rendered so much more distinct. But few vibratile epithelium cells 
 are found, and these are generally of medium size, while some are 
 very minute. They, specially at the beginning of an attack of 
 coryza, show a rapid, persistent, vibratile motion ; a large number 
 of pavement cells of different sizes also appear. Some few red 
 blood corpuscles are met with, and of course many foreign particles 
 inhaled with the air. There is no trace of myelin. 
 
 Quite a large number of the dumb-bell-shaped bacteria are to 
 be seen, as well as many of the types shown in a, b, d, and e, 
 though larger than those figured in the plate. Above all the 
 oblong forms, q and z are prevalent, some isolated, others m 
 colonies or masses, many adhering to the epithelium pavement 
 cells Some of these oblong forms have a swollen extremity, 
 resembling the Vibrio rugula. Other minute bacteria are usually 
 
48 SPUTA OF WHOOPING-COUGH. 
 
 present, more especially on the mucus corpuscles, and from these 
 minute bacteria are derived those of the beaded type, s. There 
 are, in moderate numbers, bacteria after the type of those shown 
 in / (below), but without any apparent capsule, as also various 
 diplococci with apparent capsules, as those shown in /'. These 
 last were first noticed by Thost* and Lowenberg, in 1886, as will 
 be seen from the Fig. 23 in the quoted work on Microscopical 
 Diagnosis by Jaksch.f 
 
 Lastly, some few specimens of a rare bacterium, of the straight 
 type, />, /, are found in couples, and now and then bacilli, quite 
 small and very slender. In no single case did I notice in healthy 
 mucus the Sirepiothrix Forsteri, but once found it in a rhinolitic 
 concretion, having as a nucleus a cherry-stone, which had been 
 buried in the right nasal fossa for a whole year. 
 
 Bacteria and Bacilli found in the Sputa in Bronchitis. 
 
 In the clear mucus, during acute attacks of bronchitis, the 
 bacteria are very numerous, accompanied by a few mucus corpus- 
 cles, together with some salivary corpuscles and buccal epithelia, 
 a large number of pavement epithelium cells from the air-passages, 
 as well as some few vibratile epithelium cells from the same, and 
 in some instances by some red blood-corpuscles. 
 
 The bacteria found are those of the types «, b, c, d, <?,/, i, k, I, 
 m, n, p, g, s ; to these may be added masses, as shown in u, u, of 
 small slender bacilli, spirilli, and filaments of Leptothrix, etc. I 
 must also draw attention to the fact — not noticed, I believe, by 
 other observers — that the mucus corpuscles are at times invaded 
 by minute diplococci, occasionally free, but sometimes apparently 
 enveloped in a capsule. Such a fact is of no less importance than 
 that which was held to be a characteristic of inflammation of the 
 lungs — viz., the formation of numerous diplococci within mucus 
 and pus corpuscles. Besides the imprisoned ones, others are to 
 be seen with a semblance of a capsule, such as are shown at f, 
 l\ and p'. 
 
 * Thost, Deutsche Medicinische Wochenschrift, 1886, 12, 161. Lowenberg, 
 ibid., 446. 
 
 t Jaksch, Manual (quoted), p. 58, Fig. 23^. 
 
sputa op whooping-cough. 49 
 
 Bacteria and Bacilli found in Pneumonitic Sputa. 
 
 The same variety of types just enumerated are found in the 
 sputa in cases of inflammation of the lungs, but the types with 
 apparent capsule seem relatively to predominate. 
 
 The straight forms, as /, /, sometimes free, at times enclosed 
 within a transparent envelope, and now and then covering and 
 invading such buccal epithelium cells as are breaking up ; their 
 numbers increase day by day, especially in hot weather. The 
 beaded forms, s, and the matted groups, u, u, are present. 
 
 The forms with clear surrounding matter vary in nuhibers in 
 different cases, but never seem to predominate over the other 
 forms. They oftentimes are mixed in the same group or mass 
 with bacteria of different types. These, too, increase in numbers 
 in stale sputa. We find also the forms, as in the lower part pf 
 Fig. /(biscuit-shaped), those as shown in /' on the right, the forms 
 e andy^ and that shown in /', made up of four rounded bacteria 
 or cocci strung together. 
 
 The apparent capsule is not merely the outcome of some opti- 
 cal illusion caused by contraction of the surrounding medium, 
 leaving a vacant space, for such capsules are equally visible in 
 specimens contained in newly added liquid medium, beyond the 
 striae of the mucus ; and, moreover, some of the bacteria pre- 
 senting such apparent capsules — those of type /', — are endowed 
 with a somewhat lively motion, and still carry with them the same 
 appearance. But, still, there is not usually any distinct sign of a 
 true capsular membrane or sac. It seems to me, therefore, that 
 we have here simply a clear, transparent matter, not acted upon 
 by any dyes, surrounding the bacteria, or else cilia of the most 
 minute kind. This last supposition will explain the motion of 
 these forms and the difficulty of staining the apparent capsule ; 
 but no one appears to have observed such cilia, even under the 
 highest powers.* 
 
 * On this question of vibratile cilia, see, among others, Van-Tieghem, Sur 
 les pritendus His des'bactiries. Bulletin de la Soc. Botanique, Paris, 1880. As 
 regards the flagella of Bacterium ternio, see Dallinger and Drysdale On the 
 existence of flagella in Bacterium termo. The Monthly Microscopical Journal, 
 1875. Cornil and Afanasieff attributed the capsular appearance to the con- 
 traction of the medium (Lemons & la Faculti, etc., 1883—84) ; but Cornil seems 
 
 B 
 
50 SPUTA OF WHOOPING-COUGH. 
 
 In my researches I have found that the capsular appearance of 
 these bacteria is rendered more distinct by the use of methyl violet 
 than by gentian violet. This may arise from the methyl violet 
 staining more deeply and with a bluish tint both the bacteria and 
 the surrounding mucus, thus showing up more distinctly the 
 intervening and unstained zone. 
 
 All these capsular bacteria, but principally the smaller ones, 
 invade the buccal epithelium cells in pneumonitic sput£i, but in 
 fresh specimens they are difficult to perceive within pus corpuscles. 
 
 In some pneumonitic sputa I have noticed the beaded forms, 
 as in ic' (x 1,250 diam.), where will be seen two rounded bacteria 
 or articulations, with three others, elongated, at each extremity of 
 the chain. These elongated forms in no way differ from those in 
 q and x (above) and z ; the difference of scale in the drawings 
 should be borne in mind. This tends to establish still more the 
 identity of the round and elongated forms. 
 
 In other pneumonitic sputa, examined at the same time, I have 
 observed innumerable and most minute bacteria arranged in line 
 among th,e striae of the mucus as a string of beads. I believe 
 them to be identical with the forms e, f, and / (the straight speci- 
 mens), but younger in growth. 
 
 Examining the sputa on the following day, which was a very 
 warm one, I found these minute bacteria had increased in greater 
 numbers than any of the other forms. Slender threads of growing 
 bacilli, as V, were also seen ; among such chains as are seen in c 
 were notjced, with a greatly increased number of the straight bac- 
 teria, some of the type/. The invasion of the buccal epithelium 
 cells by the bacteria was more and more evident. 
 
 In a case of inflammation of the lungs, the temperature being 
 30'C, some sputum was preserved in a closed glass tube, previously 
 cleaned with sulphuric acid and rinsed in alcohol. On the fourth 
 day the material turned quite black. I found this change was 
 caused by the buccal epithelium cells turning to a clear brown, 
 
 to have modified his views. See, too, Dallinger on The Cilia of Bacteria ; 
 Carpenter, The Microscope and its Revelations, 7th ed., London, 1891, p. 587.. 
 The researches of Loeffler and others on the staining of these ciUa are well 
 known. As to the apparent capsule, the succeeding memoirs may shortly be 
 referred to.~(F. V4) 
 
SPUTA OF WHOOPING-COUGH. 51 
 
 and by the nuclei and granules taking a still much darker colour, 
 which had also spread in a lesser degree to the other morphologi- 
 cal contents. The sputum also contained a large number of red 
 blood-corpuscles. It seems to me not at all improbable that some 
 tannate or malate of iron had been formed by the mutual action 
 of hsematin and some tannic or malic acid, which could be ac- 
 counted for by the patient having eaten some unripe grapes or seeds 
 of pomegranate or similar fruits. The patient was a farm labourer. 
 
 I cannot believe any of these bacteria can have any specific 
 character, whilst even the so-called pneumococci, with apparent 
 capsule, are not exclusively found, either in pulmonary inflamma- 
 tion or pleurisy. It has certainly never as yet been proved that 
 the germs of the bacteria with apparent capsule do not owe their 
 origin to identical forms found usually in saliva ; and what is more 
 urgent still is, that I have found these identical bacteria, as I men- 
 tioned before, within corpuscles of the bronchial mucus, in urine, 
 and in the spermatic fluid. A youth suffering from spermator- 
 rhcea, emitted in every defecation some of this fluid, and on this 
 being examined an abundance of spermatic filaments in lively 
 motion was found, together with a few pus corpuscles and mucus. 
 On this material being stained when fresh with methyl violet, I 
 found present some of the dumb-bell bacteria, and not a few of 
 the capsular bacteria of the type of those in /' and/'. 
 
 I am not pretending to bring this forward as a new discovery, 
 for such apparently capsular bacteria and diplococci, morphologi- 
 cally identical with the so-called pneumococci, have been observed 
 by others in many different substances. Taiamon went so far as 
 to say that all those microbes that are found in bronchial mucus can 
 assume a capsular appearance.^' 
 
 It is well known, too, that the diplococci, whether capsular or 
 not, which infest the blood, spleen, and liver in salivary septicaemia 
 (an experimental disease produced by the subcutaneous inoculation 
 of rabbits with pneumonitic sputa) cannot be differentiated from 
 analogous forms found in the saliva.t 
 
 * Comptes-rendus de I' Acad, de Midecine, 1886. 
 t Some attribute the discovery of salivary septicaemia to Fraenkel ! but 
 according to Cornil and Babes (pp. eit., p. 187), Klein appears to be the first to 
 have inoculated rabbits with sputa from patients during inflammation of the 
 lungs of the human subject. 
 
52 SPUTA OF WHOOPING-COUGH. 
 
 As far back as 1885, we find that Fraenkel and Platonow 
 observed this semblance of a capsule in the most varied forms of 
 bacteria found in sputum,* and some few months later Sternberg 
 also observed the capsular appearance in certain bacteria of the 
 saliva.t Previously, Thost had described some capsulated diplo- 
 cocci and a diplococcus identical with that of Friedldnder, v?hich he 
 observed in nasal mucus, both in the healthy state and in cases of 
 Rhinitis and Ozaena.J 
 
 More recently Netter noticed in the mouths of healthy persons 
 microbes identical with those observed in pulmonary inflammations 
 and in salivary septicamia. § 
 
 Though it is true that the forms with capsule never seem to be 
 so abundant in other cases as they are in cases of pneumonia and 
 pleurisy ; yet this may depend upon the peculiar condition of the 
 secretion in the air passages and bronchial tubes in these diseases, 
 which may favour their development and increase. || 
 
 Bacteria and Bacilli found in the Sputa from Pleurisy" 
 
 In genuine cases of pleurisy, where there is no collateral hyper- 
 aemia, the' sputum is poor in the matter of morphological elements ; 
 it contains only a few pus corpuscles, and the red blood-corpuscles 
 
 * Deutsche Medicinische Zeitung, Feb., 1885. 
 
 •^American journal of Medical Science, July, 1885. 
 
 X Berliner Khnische Wochenschrift, 1884, No. 25, p. 388. 
 
 % Bulletin Midical, May, 1887. 
 
 II Hunter Mackenzie denies the specific character of the pneumococci, and 
 quotes a case in which an unusually large number of these bacteria were found 
 in the sputa ot Hydrothorax. In a case of traumatic pneumonia, through the 
 fracture of the fourth right rib, the sputa showed to me for many days an abun- 
 dance of pneumococci. In cases of pneumonia supposed to be infectious, I have 
 found these pneumococci in larger numbers during the latter period of the 
 malady (as in the third week) than at its beginning ; surely, on the supposition 
 that such forms have a specific character, the contrary should take place. 
 Lately, Sternberg (The Medical Record, March, 1889, Nos. 958 — 59) acknow- 
 ledges their buccal origin, and attributes their lodging themselves within the 
 lungs to a peculiar disposition arising from the very cause of the malady. 
 
SPUTA OF WHOOPING-COUGH. . 53 
 
 are frequently entirely absent. Where the fever is high and the 
 difficulty in breathing acute, these capsular bacteria are even more 
 numerous and larger than in pneumonia, as shown in f on the left 
 side of the plate and in / below ; the quick and frequent inspira- 
 tions carrying the germs from the nQse and mouth will account for 
 this. There are found in this sputa the dumb-bell form and 
 others — even those of the type /,/, gathered often round the 
 masses or tufts of Leptothrix when presept. 
 
 On the third day, in a case of pleurisy with effusion, fresh 
 sputum was treated with methyl violet, and was found to contain 
 the bacillus drawn in Fig. 2, «' ( x 2,500 diam.). The two ends of 
 the bacillus are deeply stained and opaque, whilst the inter- 
 mediate part is clear but granulated, and contains minute bacteria, 
 some placed lengthways and others across the axis of the bacillus. 
 This specimen may be taken as an example of transition of the 
 younger forms, n, n, to the maturer forms, m, m, here evidently 
 showing a stage of the transition. For in n' the two extremities are 
 exactly similar to the forms m, m, .whilst the middle part retains 
 its.likeness to that seen in n (below, on the left). There can be 
 no doubt as to the identity of these two forms. 
 
 These bacilli of the type «', and similar ones, surrounded a 
 new formation of a tuft of Leptothrix, or really formed a part of 
 it ; and we may safely infer that the types n', n, m, u, s are the 
 same as q, if, and z ; and these, again, are the same as x', x, y, 
 and r. 
 
 Judging from the types shown in the plate and alluded to in 
 the text, we may conclude that all may be reduced to two species: 
 Bacterium termo amd Leptothrix. We can refer the forms a, b, c 
 (above), d, e,f,f, g, i, k, I, l,p, and / to Bacterium termo, and 
 1 the forms V, c (below), m, n, n', o (on the left), q, r, s, u, v, 1/, x, 
 x', y, and z to Leptothrix. 
 
 Nor is it improbable that even these two forms are one and 
 the same species. Cohn, Naegeli, De Bary, and others hold that 
 Bacterium termo, Leptothrix, and the round bacterium {Monas 
 crepusculum) are but three forms or stages of one and the same 
 species. I am also of opinion that the forms known as Vibrio 
 
54 . SPUTA OF WHOOPING-COUGH. 
 
 rugula, Clostrium, and Rhabdomonas are of the same species, 
 being varieties of the same form of bacilli.* 
 
 Bacteria and Bacilli found in the Sputa in Phthisis. 
 
 I can but merely touch on the matter of bacteria in phthisis, 
 as it presents a field far too wide to be treated adequately in the 
 short space of this memoir, and for the same reason I shall not be 
 able to enter into a discussion of the methods of culture or 
 inoculation. As my observations were only of a semiological . 
 character, and although I do not pretend to offer the results of a 
 fully scientific research, I am of the firm opinion that the morbific 
 character of bacteria cannot be fully established until the natural 
 history of micro-organisms, in part at least, be first ascertained, 
 and this, I believe, has yet to be done. This much is certain : 
 that bacteriological research cannot be considered an infallible test 
 in the diagnosis of disease, nor, without caution, can it be consid- 
 ered as pathognomonic in clinical medicine, until in daily practice 
 it is found to tally with the other observations made during the 
 course and termination of disease. 
 
 Having made this statement, and reserving for another place 
 some statistics on the question, I will now proceed to offer an 
 objection to the assumption of the existence of any tubercular or 
 phthisical bacillus in sputa. 
 
 All the fervid supporters of this bacillus and its pathogenical 
 power have arrived at the conclusion that its morphological cha- 
 racters are dubious and of little value, and that no bacillus can bS 
 assumed to be the tubercular bacillus, unless when stained with 
 gentian violet it is not decolourised, although treated with a mix- 
 ture of dilute nitric acid and alcohol. Without this test, they say, 
 no bacillus can be held to be the bacillus of tubercle, even if its form, 
 length, and disposition be the same as that of the type described. 
 
 * For the sake of brevity, I have omitted the distinguishing characters of 
 the pneumococci (Friedlander's and Fraenkel's, etc.). Most likely, the differ- 
 ences observed are but accidental, and may arise, I think, from different 
 degrees of staining imparted to this or that portion of each member of a 
 microbe, to the point, side or connecting matter. Anyone can assure himself 
 of the numberless forms, too many to describe here, found in any one mass of 
 pneumococci. 
 
SPUTA OF WHOOPING-COUGH. 55 
 
 These are the words of my esteemed friend, Prof. Primavera, a 
 most strenuous upholder of this bacillus.* 
 
 The only distinguishing test between the bacillus of tubercle 
 and other bacteria or bacillus in sputa is a relative difference in 
 their staining power, or, in other words, a difference in their che- 
 mical value. I call it a chemical test, as the process of drying, 
 the colourising with aniline dye, and the warming over the lamp 
 must destroy all the vital properties of the bacillus, t Gentian 
 violet is a basic stain, owing to the amount of oil of aniline it con- 
 tains ; it is readily soluble in alcohol, as are all other aniline dyes. 
 The test, therefore, resolves itself into the question as to whether 
 the affinity of the bacillus to retain the stain is stronger than that 
 of the power of the acid and alcohol to remove it. 
 
 In other words, if the staining of the bacillus be only a che- 
 mical phenomenon, the decolourisation, too, must be of the same 
 nature. So that the taking up and retaining the colour on the one 
 hand, and parting with it on the other, will depend upon condi- 
 tions altogether fortuitous and variable, and which must be of a 
 purely chemical nature. Now, a purely chemical characteristic 
 can hardly be elevated into a morphological or biological one, fit 
 to determine a particular species of micro-organism. At the 
 very most, it might be used as a criterion in diagnosis ; but, even 
 reduced to this, can we be sure that this criterion is not fallacious? 
 
 You may heat the preparation a little below or above the 
 proper temperature ; you may add a little too much or too little of 
 the stain ; you may apply it while the preparation is warm or cold ; 
 you may allow the dye to act for too long or too short a time ; 
 you may use a stain which is quite fresh or it may be old ; you 
 may allow the nitric acid io act a second too long or too short; you 
 may, when washing the preparation in alcohol, allow the violet or 
 green tint to be visible to the naked eye, or by the addition of 
 more alcohol destroy the tint altogether. In each of these differ- 
 ent suppositions you will necessarily obtain different results. In 
 all probability, sometimes the results will be satisfactory in a 
 
 * Primavera, Manuale di Chimica e Microscopia Clinica, Nap&li, 1887—88, 
 
 p. 400. 
 ilbidem, p. 399. Relating to the poisonous action of aniline oil on the 
 above bacilli. See G. Sormani, Nota al R. I. Lombardo, 15 Dec, 1887. 
 
56 SPUTA OF WHOOPING-COUGH. 
 
 practical sense ; but when the microscopical examination is 
 systematically deferred till the decolourisation with nitric acid and 
 alcohol is complete to the naked eye, or, what is more important, 
 until the alcohol used in washing is completely colourless as it drops 
 from the slide, the reliable results will be considerably lessened 
 and your faith greatly shaken in this distinguishing test. 
 
 It is some time since I noticed this difficulty, and only lately I 
 happened to come across an observation of Jaksch to the same 
 effect. This author recommends that the dilute nitric acid and 
 alcohol should be allowed to act only long enough, to change the 
 violet tint to green. " C« ne saurait," he. adds, " consei liter 
 d'attendre que les preparations soient complktement decolorkes, car, si 
 on attend trop long temps, sous Faction knergique de Vacide, les bacilles 
 (de KocK) finissent par se d&colorer." * 
 
 Another source of error ' is the unequal distribution of the 
 sputum on the slide ; but I must now leave this question. Nor 
 can I now give an account of the technique in use, but must 
 reserve the matter for another time. 
 
 One thing, very evident from these bacteriological observations 
 is, the very great affinity which the beaded forms of the bacilli in 
 question have for gentian and methyl violet. But those who bear 
 in mind the various grades of tint imparted to the bacteria by 
 these stains will not be at all surprised, for we nearly always find 
 that the more minute bacteria in sputa take the stain more tena- 
 ciously, with the exception of those more fully developed, which 
 are enveloped in some sort of cuticle (as we saw in the case of 
 the forms i and k) ; in such the opposite takes place — the larger 
 and full-grown specimens are the more strongly tinted. Now, the 
 individual bacterium or articulation that forms the beaded type, 
 J, is the smallest met with in sputa, and it is not unlikely that 
 
 * Jaksch, Manual, p. 73. It is well to notice that the Gratz' Professor, a 
 strong upholder of the value of this bacillus in sputa in diagnosis, uses methyl 
 violet. Hunter Mackenzie does not believe in the selective power of this 
 bacillus for gentian violet, preferring fuchsine ; but this even disappears si le 
 bain cFacid nitrique est trop prolong^ (more than 30 sees.) ( V., loc. cit.,p. 141). 
 Lately, carbolated fuchsine has been recommended ( To Stain Tubercle Bacilli, 
 in The Journal of Microscopy and Natural Science, edited by A. Allen, July, 
 1889, pp. 165 — 66), and which answers well in practice. 
 
SPUTA OF WHOOPING-COUGH. 57 
 
 for this reason alone, or on account of the enveloping cuticle, and 
 not through any biological faculty, they are able to take up the 
 dyes more energetically. 
 
 But there is still more to be said on this subject. This power 
 of retaining the dye does not exclusively belong to the bacillus of 
 Koch, as I have satisfactorily proved in my own experience. 
 
 In the first place, it is not an unusual thing, in the examination 
 of phthisical sputa, which has been stained when dry with gentian 
 violet and decolourised with nitric acid and alcohol, to find a 
 small foreign body, such as a shred of minute membrane, or 
 frustule or filament of vegetable origin, or some epithelium cell 
 which still retains the blue tint. Then if such foreign body is 
 covered with, or retains any bacteria, these will be found- un- 
 bleached, though they are of a type altogether different from that 
 of Koch's bacillus. These bacteria are extremely minute, yet 
 after the type of those in /' and p\ usually found in the sputa 
 under consideration, not only when fresh, but even after treatment 
 with potash, as I shall presently show. 
 
 In the second place, we not infrequently find in such dry 
 preparations, treated with gentian violet and then decolourised, 
 colonies of minute capsular diplococci, of the types /', /', and /', 
 sometimes singly, or two, three, or four together in one capsule. 
 These colonies alone retain a distinct, though faint, tint or shade 
 of colour, which allows the individual members to be made out 
 when mounted in a clear, refractive mounting medium (oil of 
 cloves or glycerine) ; while all the other forms, not including those 
 of the beaded type, s (Koch's bacilli), are bleached, and conse- 
 quently invisible. 
 
 From this it is evident, not only that many bacteria of various 
 types retain the gentian violet stain, but even do so at times in 
 higher degree than the so-called tubercle bacillus. I have 
 observed many striking examples of this in pneumonia. The 
 specimens, which were from sputum extraordinarily rich in pneu- 
 mococcus, had been dried and stained with gentian violet. Any- 
 body can verify their resisting power to a second altevipi to bleach 
 them with the usual dilute nitric acid and alcohol mixture, though 
 the retained tint is faint. 
 
58 SPUTA OF WHOOPING-COUGH. 
 
 Nevertheless, it is not my intention either to impugn or affirm 
 the existence of the bacillus of tuberculosis, but simply to raise a 
 reasonable doubt on this most important question of clinical micro- 
 scopy, which is still sub judice. The judges of the matter in the 
 first case may be the band of technical investigators, but the 
 ultimate and definite decision must be left to clinical practi- 
 tioners. 
 
 One of the most striking arguments to support the idea of 
 infection or contagion by bacteria is the diffusion and reproduction 
 of these micro-organisms through the tissues in different parts of 
 the body. Although this question only touches us indirectly, I 
 would observe that all living material, whether healthy or in a state 
 of decomposition, is liable to this whenever the parts are suffi- 
 ciently attenuated to allow the particles to pass from their first 
 nidus to parts more or less deep, so we must not be surprised to 
 find in autopsies that the tissues have been invaded by bacteria, 
 and especially by those that thrived at the seat of the disease. 
 There is no reason for urging this as a proof of the morbigenous 
 power of the organisms in question. In fact, the very opposite 
 opinion that such bacteria are harmless in general can offer a very 
 simple and plain explanation from the alteration of the affected 
 parts. It is but a general law, which holds good both in a 
 healthy and a morbid state, that the products of disassimilation in 
 the higher organisms become, ipso facto, the source of assimilation 
 in micro-organisms ; and the reason is this, that bacteria deprived' 
 of chlorophyll are incapable of decomposing the anhydrous car- 
 bonic acid of the atmosphere, and their growth is carried on at 
 the expense of higher organisms. This law accounts for the 
 diffusion and the increase of micro-organisms wherever particles of 
 matter are found in disintegration or in any part removed from 
 vital influence, the sovereign power in higher organisms. In all 
 cases where such alteration of living matter is in progress, and 
 where the bacteria or their germs are not excluded by special 
 and exceptional means, we not only find micro-organisms, hut they 
 are necessarily there, and cannot by any possibility be absent. 
 
 Further remarks on this point will be found in a previous 
 communication of mine on Relapsing Fever, where I have 
 
SPUTA OF WHOOPING-COUGH. 59 
 
 brought forward other objections to the supposed power of bac- 
 teria in communicating and spreading disease* 
 
 I think, moreover, that the relative predominating number of 
 the beaded bacilli, s, can be explained without having recourse to 
 the assumption of their power to originate disease. During the 
 chronic process of disintegration in the pulmonary substance, the 
 morbid products will naturally be more frequently, and for longer 
 intervals remain, lodged in cavities, not being carried to other points 
 by the currents of air in respiration. This quiescent state tends 
 to the further decomposition of the morbid secretions, and we 
 have seen in the second section {Sputum in Pneumonia, July 31), 
 and in the fourth section {Bacteria, etc., in Healthy Sputa), that in 
 sputa undergoing decomposition the more minute forms of bac- 
 teria are predominant. In the second place, this quiescence and 
 immobility of the menstruum, or nourishing medium, must neces- 
 sarily favour the juxtaposition and formation of the single bodies 
 in lines or threads, then to their multiplication and increase in 
 linear series, and eventually to the formation of beaded strings, 
 and from that to rods or filaments. This can be verified in all 
 kinds of sputa (not excluding healthy sputum) whenever they 
 have been closely kept in vessels, and day by day the beaded 
 bacilli may be seen to'lhrive and multiply, together with the inter- 
 woven growths of bacteria. 
 
 Returning to my own observations, I maintain that the bac- 
 teria found in phthisical sputa, which has been stained when fresh, 
 does not substantially differ from that found in other sputa beyond 
 the numerical predominance of this or that type. In some the 
 form s may abound, and in other specimens it may scarcely be 
 seen. The interwoven forms — as u, u — may at times be observed, 
 more especially when the larynx is affected, probably .on account 
 of the ease with which morbid secretions can be lodged for some 
 time in the folds or recesses of that organ. In dry preparations 
 
 * So'pra un caso di febbre ricorrente e sul risconfro degli sfirilli nel sangue, 
 Atti R. Accad. Med. Chir. Napoli, Vol. xxxvi., 1883. I would remark in 
 this matter that Ebstein gives an account (Berliner Klinische Wochenschrift , 
 31, 1887) of a case of chronic relapsing fever with nine relapses and a total 
 duration of 211 days, a case analogous to one described by myself four years 
 before, but in which there were six relapses in a course of 94 days. 
 
60 SPUTA OF WHOOPING-COUGH. 
 
 the beaded forms j seldom resist the decolourising action of nitric 
 acid and alcohol. But any conclusion arrived at in this way, as 
 to the nature of the bacteria, is but a doubtful one. Any two 
 observers may easily arrive by it at very different conclusions ; 
 hence, unfortunately, such conclusions as may be arrived at cannot 
 be relied upon in judging of the state of a patient. I am of 
 opinion that of the bacilli in sputa, those showing a reticulated 
 structure (they are unbleached by the usual reagent) are a much 
 better guide in the diagnosis of phthisis. If any be taken as such, 
 I, however, take them to be simply an unstudied form of Leptothrix. 
 
 In this sputum the dumb-bell bacteria are found in profusion, 
 unchanged in form, and groups and colonies of considerable size, 
 even in the sediment from sputum which has been treated with an 
 emulsion of potash. Both the free and capsular forms of bacteria 
 of the type /, / are also very abundant. 
 
 Those apparently capsular bacteria of' the type p' are quite 
 visible in the sediment from emulsionised masses of sputa, even 
 when they are mixed with other minute forms of bacteria or dip- 
 lococci, surrounded with the same kind of clear capsule, and 
 which, though of smaller dimensions, resemble the forms of the 
 types p', f, and I'. 
 
 It may be well to state here that, after having treated and 
 heated the sputa with potash, it is my custom, when searching for 
 the elastic pulmonary fibres, to allow the sediment to settle in a 
 conical glass, and by shaking and twisting the glass to hasten the 
 descent of the tufts that might settle on the sides. When the 
 sediment has been well deposited (which may take some two 
 hours), I decant the clear, supernatant liquid, and add about as 
 much distilled water, stirring the whole with a glass rod to break 
 up the sediment. In this way it is thoroughly washed from all the 
 numerous granular particles and oily globules that would other- 
 wise spoil the preparation. If one washing is not sufficient, I use 
 another. 
 
 Preparations from these deposits are excellently stained, when 
 fresh, with gentian violet or methyl violet, rendering the beaded 
 types f, with the other bacteria and diplococci, both large and 
 small (spoken of above), very distinct, as well as the apparent 
 
SPUTA OF WHOOPING-COUGH. 61 
 
 capsular forms. Very often the above beaded forms and bacteria 
 of other types are mixed together in one group or mass.* 
 
 In the article on Sputa in Whooping-cough (see the first chap- 
 ter of this Section), I spoke of the bacillus /, c, of which I found 
 some few specimens in this class of sputa. 
 
 Summing up what I have hitherto observed, my opinion is that 
 the character of the beaded bacillus of the type s is altogether 
 accidental ; while the contrary opinion respecting their specific and 
 pathogenic character is confronted with two very grave objections. 
 Firstly, that these beaded forms are found to exist in other kinds 
 of sputum, including that of healthy subjects. To this objection 
 it may be answered that the beaded forms are spurious when they 
 do not resist the decolourising action of the nitric acid and alco- 
 hol. And secondly (and which is a more serious objection), these 
 beaded forms, also from phthisical sputum, abound, not only 
 surrounding, but actually within, the very body of the buccal epithe- 
 lium cells in the process of breaking up. Some of these cells are 
 literally packed with the bacilli. This can hardly be explained 
 except by granting that the origin of such bacilli is to be found in 
 the mouth. t 
 
 Respecting the diagnostic value of bacteria in the examination 
 of any kind of sputa, I maintain that it is quite impossible to 
 
 * This method of examination for Koch's bacilli in sputa treated with an 
 emulsion of potash is of my own devising. I find that Biedert uses a modifica- 
 tion of this method when staining and examining the sputa in its natural state. 
 He adds a little fresh egg albumen, and then stains a portion of the albuminous 
 deposit. (Journal of Micro, and Nat. Sci., N.S., Vol. II., p. i66, July, 1889, 
 loc. cit.) 
 
 tin a resume of a lecture from Sternberg at Albany, written in February, 
 l88g (four months after my communicating the results of my researches to the 
 Royal Academy of Naples), I find the following quotation, which, though sus- 
 taining other views, seems to predict the fact, already observed by me, that 
 Koch's bacilli might be found in buccal epithelium cells, even under normal 
 conditions : — " It is quite probable, in the present state of our knowledge, that 
 the tubercular bacillus may live in the buccal mucous membrane, in that of the 
 nose, or even in the larger bronchi of those persons living in crowded districts, 
 and that it then attacks the pulmonary tissue when this last is predisposed, 
 in an unhealthy condition, or so injured as to present a suitable medium for its 
 development." — The Med. Record, 1889, Nos. 958—959. 
 
62 SPUTA OF WHOOPING-COUGH. 
 
 ascertain from any baderioscopic investigation whether the sputa is 
 from a healthy or from a diseased subject ; or, under the second 
 hypothesis, what is the nature of the disease ? There may be 
 exceptional cases — sometimes in tuberculosis, pneumonia, and 
 pleurisy — to this general rule, as in such cases of tuberculosis 
 where the beaded forms, s, by combination, resist the -decolouris- 
 ing medium, or are accompanied by reticulated bacilli j and in 
 those cases of pneumonia and pleurisy, in which the forms of 
 bacteria or capsulated diplococci are in such quantities as to out- 
 number all other bacteria present. In all other cases, if any 
 certain conclusion is arrived at, it will never be from merely bacteri- 
 oscopic examination, but from other characters of the sputum, 
 
 § V. 
 OBSERVATIONS AND CONSIDERATIONS ON THE FUNGI. 
 
 As I before stated, I have observed two kinds. of fungi in 
 sputa of Whooping-cough. One, the most conspicuous, was met 
 with in three cases out of the fifteen. In the first of- these cases, 
 it was found in all the sputa examined up to the end of the attack. 
 It was not found at all in the pulmonitic sputa of the patient after 
 his recovery from the Whooping-cough. The other fungus, which 
 was much smaller, was observed only in one case ; but it might 
 have escaped my notice through want of time and patience in 
 searching for it. 
 
 The larger fungus was formed of very distinct tufts of filaments, 
 freely germinating in the sputa from the day after its collection. 
 In Fig. 3, d, is shown the middle portion of one of the tufts 
 ( X 800 diam.). It was stained a pale blue by gentian violet ; but 
 the single cells are drawn somewhat more oval in the figure than 
 they were in the specimen. The mother cell of the tuft is seen at 
 e ; on the left will be noticed a smaller cell, from which arise the 
 base of two filaments, partly seen in /and g. Between the cell 
 and the base of the filaments there was a small granular mass 
 more deeply stained than the rest. On the right of the 'mother-cell 
 two filaments branched out directly, with the granular mass at the 
 base, as in h and i. These filaments or threads of the mycelium, 
 little by little, become more slender, and were more deeply stained 
 
SPUTA OF WHOOPING-COUGH, 63 
 
 at tHe extremities when they were treated with gentian violet, and 
 even more distinctly so when treated with carmine. This is 
 observed in all such tufts, and arises from the germinal matter in 
 activity being so much denser at their extremities. 
 
 As the second fungus does not present any striking peculiari- 
 ties, I need not dwell upon its filaments. 
 
 Many vesicular gemmiiles, sacculi, or sporidia were given off, 
 as I have said, from these larger tufts of the type d, and many 
 were observed quite detached, sometimes isolated, sometimes 
 united in small groups. Two of these are shown in b, b, as they 
 appeared in an ordinary preparation, stained with gentian violet, 
 and magnified 2,500 diameters. To better observe the interior 
 structure, I stained a suitable specimen of sputum with Beale's 
 carmine. My method was, first, to add a drop of diluted acetic 
 acid to a particle of the sputum, and, then place it in a watch-glass 
 containing the carmine stain, taking it out the moment it appears 
 to be stained sufficiently, otherwise the tint becomes much too 
 diffuse. One of these gemmules is shown in c,* stained in this 
 manner with carmine under the same power as b. It will be seen 
 to present a lightly stained exterior layer, and an interior one con- 
 taining granules more deeply stained. These granules are more 
 vividly stained within the narrow and pointed neck, where the 
 two layers or coats seem to run into one another. Probably a 
 new growth of gemmules begins at this point, and at the same 
 time new tufts are reproduced. The contents seem to be a 
 hoiiiogeneous matter, in which are suspended some masses of 
 germinal matter and granules darker than itself. 
 
 To obtain the fructification of the fungi in question, I pre- 
 pared a culture on the inner surface of a fresh piece of lemon 
 peel by placing on it a little of the mucus which was rich in the 
 tufts, and enclosing it in a glass vessel, which was then placed in a 
 dark cupboard. Another culture was tried on potato and flour 
 paste without any result. Perhaps these substances dried up too 
 quickly at a temperature of 22° C. The first culture was examined 
 every morning. On the fourth day I found it covered with mould 
 in greenish patches, from which arose little dark heads, quite 
 
 * This figure should be carmine, but has been tinted violet for 
 purposes of illustration. 
 
64 SPUTA OF WHOOPING-COUGH. 
 
 visible under a single lens. The greenish patches were the smaller 
 fungus ; the small dark heads the larger one. After two more 
 days' growth I ?liced off the aerial hyphag with a scalpel, placed it 
 in a drop of glycerine on a glass slide, and applied a thin cover 
 glass supported by two slips of paper, to prevent" the little heads 
 from being crushed. 
 
 The fructification of the larger fungus — which, so far as I can 
 judge, belongs to the genus Mucor — is shown in its natural state, 
 k and k\ being mounted in glycerine and magnified to only 140 
 diam. The reproduction of the drawing of k is somewhat imper- 
 fect, owing to the difficulty of representing exactly the degree of 
 transparency at different points and the tone of the natural colour. 
 
 The hyphse of this fungus are not septate. They are slightly 
 granular towards the extremities, clear and transparent elsewhere, 
 and terminate in a rounded head, smooth and comparatively large, 
 as shown in k. This head is entirely covered by a layer of rather 
 dark spores closely packed together. Fig. k' shows some of the 
 spores still attached, while the rest have fallen off. The Fig. k 
 shows the layer of spores intact and enclosed by a thin, darkish 
 envelope, which barely allows the spores to be seen at all. It is 
 somewhat more easy to make them out at the edges, and still 
 more so at the point of attachment to the stem, where the enve- 
 lope seems to be thinner. The detached spores are shown in / 
 ( A 1,000 diam.), somewhat shrunken through exosmosis under the 
 action of glycerine. Probably the fungus noticed by Tschamer, 
 as I mentioned in the first paragraph, belongs to this species.* 
 
 The fructification of the more slender fungus is very much 
 smaller, and shown in n, 0, and /. The preparation was mounted 
 in the same medium, magnified to 800 diameters. The hyphse of 
 this fungus are not septate, and give rise at their extremities, 
 which are swollen, to a series of sterigmata or ramules, bearing 
 
 * I refer this fungus to the genus Mucor (stolonifer ?) from the following ^ 
 description given by J. Payer : — " Mucor, Micheli. Flocci erecti, simplices, 
 conlinui, afice thecam subglobosam, eyatho laceram gerentes. Columna cylin- 
 driea vel ovata. Spora simplices" in his Botanique Cryptogamique ou Hist, 
 des Families Natur. des Plantes Infh'., Paris, 1868, p. 85. As regards the 
 vesicular gemmules of Mucor, see Cooke, Les Champignons, sous la direction 
 de M. /. Berkeley, Paris, 1875, pp. 51, 52. 
 
SPUTA OP WHOOPING-COUGH, 65 
 
 an erect string of spores. The fructification in n is scarce, there 
 being but four ramules; in o it is abundant, although I should add 
 that the specimen shown at o had lost many of the spores in the 
 upper part of the branches, for in many other specimens the fan- 
 like pencils are three or four times as long, contracting as they 
 rise and ending in a plain horizontal section, as is often observed 
 in Aspergillus nigricans. In the specimen shown in Fig. p, the 
 fructification is covered with a dark, opaque veiling, through 
 which only the marginal spores can be seen ; but in many speci- 
 cimens the veil only partially envelopes the spores, sometimes at 
 the lower parts and sometimes longitudinally. So it would seem 
 that the various phases of fructification denote no specific differ- 
 ence, but are those of one and the same species. 
 
 This fungus may be Aspergillus fumigaius or a similar species, 
 or perhaps it belongs to the genus Rhodocephalus.* 
 
 Probably the fungi found by Ransome in the breath of 
 patients with VVhooping-cough (see first section) were the same as 
 the above. But I hardly think he can be right in describing it as 
 Penidllium, as the hyphfe of Penicillium, as well as their ramules, 
 are septated, whereas those of this fungus are continuous. 
 
 I observed in Section II. (Sputa of May loth, c) that the 
 smaller fungus, after some days, gave the sputum a greenish 
 colour. I may here add that even on the fourth or fifth day this 
 colour may be observed by transmitted light. If the sputum be 
 preserved in a glass tube, the formation of cloudy patches of a 
 light green colour will be easily noticed. Later the whole mass 
 becomes green by reflected light. I tried to discover whether or 
 no the development of these fungi correspond with any chemical 
 change in the sputum, and found that this was not the case. The 
 
 * " Aspergillus, Micheli. Receptaculuin floccosum eredum, continuum, 
 simplex, apice in capitulum inflatum. Flocci sporarum capitulum tegentes. — 
 Rhodocephalus , Corda. Receptdculum floccosum erectum, continuum, non 
 septatum, apice in ramulos simplices, capitulum formantes divisum. Flocci 
 sporarum apicibus ramulorum. singulatim innati." — Payer, loc. cit., pp. 70, 71. 
 In these definitions the hyphae are sometimes called flocci, and at other times 
 called receptaculum. Flocci sporarum are the moniliform strings of spores that 
 rise from the lop of the hyphse or its ramules, or from the columna. For 
 Rhodocephalus, see Fig. 303 of the same author, showing the fructification of 
 Rhodocephalus candidus. 
 
66 SPUTA OF WHOOPING-COUGH. 
 
 sputum in Whooping-cough, when quite fresh, gives a neutral or 
 very slightly alkaline reaction ; and up to the seventh day the 
 reaction remains the same, notwithstanding the visible formation 
 of these greenish patches and the increase of accompanying tufts 
 in the preparation. 
 
 There is nothing new in this matter of specimens of vegetable 
 growth in the air-passages. Nowak says " that micromycetes 
 (specially the genera Aspergillus and Mucor) are frequently found 
 in the air-passages of birds suffering from respiratory diseases."* 
 And Perroncito, speaking of Aspergilltes fumigatus, says that 
 it has been found in the human lungs (Virchow and Pagenstecher) 
 and in the air-sacs of Otis tarda, producing these pneumonitic 
 nodules {Pneumomicosi aspergilliana). Passing over the Aspergilli 
 that are found living in the auditory canal, the same writer quotes 
 cases of Aspergillus nigricans, found by Robin in the air-sacs of a, 
 pheasant, and by Generali and others in other birds; of 
 Aspergillus virens, also found in the air-sacs of birds ; of Asper- 
 gillus candidus, found in the air-sacs of Pyrrhula vulgaris ; and of 
 Aspergillus glaucus in the air-passages of crows and other birds, f 
 
 The sputum that furnished the above specimens of fungi was 
 kept in a tightly closed glass vessel until it was completely dried 
 up. On the dried film at the bottom of the vessel there was no 
 aerial vegetation, but on the sides there were growths, white in 
 colour, of the smaller fungus, as shown in n, o, and p. I suppose 
 that these growths were caused by particles of sputum reaching 
 the sides whilst tipping the glass and extracting portions for 
 examination. 
 
 In no way do I attribute any pathological importance or spe- 
 cific power to these two species of fungus which are found in the 
 sputum in Whooping-cough. Their forms are diffused in -profu- 
 sion in the air, and in the cases mentioned they may have 
 accidentally found their way into the sputum, as they were met 
 with in these few cases only. I may remark that microscopic fungi 
 
 * Nowak, Le malattie infeitive, etc., trad, di Vesica, Naples, 1884, p. 10. 
 Payer, loc. cit., p. 60. Beale, loc. cit., p. 500. G. H. Mackenzie, Figs. J 
 and 24. Jaksch, Figs. 30 and 31. 
 
 t Perroncito, I parassili delV uomo e degli animali utili, Milan, 1882, pp. 78, 79. 
 
SPUTA OF WHOOPING-COUGH. 67 
 
 found living in sputum is not a new discovery. I have often 
 found that when pneumonitic sputum has been kept for some days 
 in closed tubes, a rather dark scum of mould rises to the 
 surface; this is not to be confused with the diffused dark- 
 ening spoken of in the fourth paragraph, for under the micro- 
 scope it proved to be Penicillium. Other tufts are at times found 
 in phthisical sputum and in the sputum of persons suffering from 
 buccal disorders, or even when wearing artificial teeth. I have 
 obtained a fungus of the type k, and observed its fructification, 
 together with specimens of Penidllium glaucum from sputa of 
 persons suffering from typhoid fever. These last are very common 
 fungi, thriving and fructifying together on common flour-paste. 
 
 As regards the spores from the cistern water, of which I spoke 
 in the third paragraph, and of which I give a drawing of a speci- 
 men when germinating in Fig. 3, a. I am in doubt whether it 
 belongs to the genus Hysterographium, of the Hysteriacece family, or 
 to the genus Septosporium of the Botrytidae* These are diffused 
 everywhere in the country, even on vine-tendrils. So far, I have 
 never observed them in water laid on in towns, but they are of 
 frequent occurrence on wall-papers in moist situations or where 
 the air is confined. 
 
 Naturally, I must leave the exact determination of the three 
 species of fungi, shown in Fig. 3, to more competent micologists. 
 
 * Hysterographium, Corda. Conceptaculum simplex, sessile, rima longitu- 
 dinali dehiscens. Thecce paraphysibus mixta, tubulosa. Spora septatte." — 
 Payer, loc. cit., p. 97. Compare with Fig. 433 of the same author, showing 
 the spores of Hysterographium elongatum : — " Septosporium, Corda. Flocci 
 erecti, ramosi, pauco septati. Sports celluloses, pedicellata " — Ibidem, p. 76. 
 Compare with Fig. 340 of the same author, showing Septosporium bulbotrichum. 
 The spores of the genus Hendersonia are longer and narrower." — Ibidem, p. 83. 
 Compare with Fig. 49, Cooke, loc. cit., p. 112. 
 
68 SPUTA OF WHOOPING-COUGH. 
 
 § VI. 
 HINTS ON THE PREPARATION AND STAINING OF 
 SPECIMENS. 
 
 Leaving to another time the determination of the source and 
 seat of the production of substances ejected from the mouth, so 
 that other substances— as mucus coming from the nose, or merely 
 from the throat, or fragments of food — may not be mistaken for 
 expectorations. I will now offer a few remarks as to my method 
 of research. 
 
 When about to examine a quantity of sputum, I collect it when 
 possible on a plate, spittoon, or glass, or I receive it on a clean 
 cloth, from which it is removed by means of a spatula to any of 
 the above vessels. At other times it was found to be sufficient to 
 collect a little in a glass tube, thoroughly cleaned with sulphuric 
 acid and washed in alcohol. 
 
 In the examination of sputa in the natural state, it will be - 
 sufficient, if a portion be taken about the size of a seed of millet, 
 no diluent should be used except a little solution of sodic chlo- 
 ride, if the sputum is proved to be too dense and tenacious. In 
 staining, even a smaller portion may be taken when the consis- 
 tency will allow of it. This small portion may be taken up with a 
 curved needle, or if too liquid a similar quantity may be transferred 
 to a glass slide by means of a pipette or dropping-tube. 
 
 Having placed the portion upon the slide, and, if necessary, 
 spread it with suitable needles, a drop of the stain is applied from 
 the end of a glass rod, unless the stain is carmine, in which case 
 another method should be followed, as below. As soon as the 
 film of the sputum is seen to be sufficiently stained, it must be 
 washed, by allowing drops of distilled water to fall on it, and 
 these drops are made to run off by tilting the slide. This must 
 be done until all granulated deposit from the dye be removed. 
 The specimen is then again spread by needles, reducing it to a 
 thin uniform film, so to say, a little less in area than the covering- 
 glass to be used. A cover-glass should be used of known thick- 
 ness, and should be gently pressed down by degrees at different 
 points with the handle of a mounting glass rod well cleaned, so that 
 the film of the specimen be rendered as thin as possible and the 
 
SPUTA OF WHOOPING-COUGH. 69 
 
 cover-glass made to adhere as firmly as possible, and air-bubbles 
 squecEed out at the same time. Without these precautions, the 
 cover-glass may attach itself to the front lens of the objective 
 when the immersion oil is used, and so render any good observa- 
 tion almost impossible, on account of the difficulty of focussing 
 and of the displacement of the specimen under the cover-glass, 
 which at times may be even lifted up from the slide by its adhe- 
 sion to the objective. 
 
 After the preparation has been examined under a dry lens, the 
 condenser is adjusted and the immersion lens substituted, each 
 portion of the specimen being successively examined. If during 
 the examination (generally a slow operation in these thin layers of 
 mucous matter) there are signs of the edges of the specimen 
 becoming dry, a small drop of distilled water, very slightly tinted 
 with the dye used, is allowed to fall on the opposite side close to 
 the cover-glass, and by means of a needle drawn up to the cover 
 and round its edge for a short distance, to render the flow of the 
 water between the glasses easier. 
 
 As regards the dyes I have used, I will briefly offer the follow- 
 ing notes : — 
 
 Gentian Violet. — I generally prefer gentian violet when examin- 
 ing sputa of Whooping-cough for the reason that it is a double 
 stain ; the filaments, particles, and granules of myelin appearing a 
 more or less deep blue, the true morphological elements and the 
 bacteria being stained violet. This double staining affords great 
 ease in observations, and moreover renders most distinct the 
 myriads of the minutest granules of myelin, which would other- 
 wise be passed over or mistaken for amorphous granulated matter 
 or for micrococci. These granules are mostly to be found, often 
 in lively motion, in those clear spaces (or little gulfs, as they 
 might be called) formed by the liquid medium flowing, as it will 
 do in these preparations,' backwards and forwards, within the spe- 
 cimen ; or else they will be found around the edges of the 
 preparation, where the granules and particles of myelin are carried 
 by the tiny centrifugal currents. So also the larger particles arid 
 masses are well marked, the gentian violet rendering them less 
 refractive j but it does not prevent the internal striae from being 
 quite visible. 
 
70 SPUTA OF WHOOPING-COUGH. 
 
 If the staining has been carried too far, the tint of the myelin, 
 after four or five hours, changes from blue to violet. Moreover, 
 the excessive stain is decomposed, and runs together in smaller or 
 larger drops of a deep blue colour, and at last they surround and 
 absorb the myelin particles, which then begin to disappear, 
 remaining only visible where the staining was more feeble. So it 
 is important to avoid staining too deeply, for this would render 
 the preparation quite useless after a few hours. As I have stated 
 before, the preparation should be carefully washed with distilled 
 water after staining. 
 
 Solution of Iodine in Potassic Iodide. — This stain, either applied 
 as above or simply allowed to run into the preparation by capil- 
 larity, gives the myelin a very brilliant yellow colour, making it 
 quite easy to recognise this substance. Pus corpuscles, bacteria, 
 and epithelium cells are either stained very feebly, or not at all, 
 with the exception of the ellipsoidal or alveolar epithelium cells, 
 which then show distinctly their myelin granules. This stain has 
 one drawback : it does not colour the thin granules and filaments 
 of myelin, which are at best feebly stained by gentian violet. 
 
 Picric Add. — The saturated solution of this acid in water 
 imparts a pale yellow tint to myelin, but renders the internal striae 
 rather indistinct ; bacteria, pus corpuscles, and epithelium cells 
 are stained to a deeper yellow. It is a useful stain, the smallest 
 particles of myelin becoming visible, as it diminishes their exces- 
 sive refrangibility, and when the simple round diaphragm is used 
 the molecular motion is most distinctly seen with it. The granules 
 of myelin are then observed as minute points, somewhat darkened 
 when out of focus, but quite brilliant when within the true focus. 
 But in a few hours the myelin begins to undergo a slow decompo- 
 sition ; it is reduced at first to mere faded fringes, spmewhat akin 
 to those produced with oleic acid and liquid ammonia, and these 
 eventually totally disappear. 
 
 Methyl Violet. — This stain renders myelin more opaque, and 
 sooner or later dissolves it into a homogeneous mass of a pale 
 blue colour. This mass adheres to any corpuscular bodies near it, 
 and takes many fantastic shapes. I have already spoken of the 
 advantages of methyl violet as a means of distinguishing the 
 apparent capsule of certain bacteria in pulmonitis and pleurisy. 
 
SPUTA OF WHOOPINQ-OOUGH. 71 
 
 Fuchsim. — This stain, either diluted or used in small propor- 
 tions, is very effectual in showing up the smaller granules of 
 myelin, as it increases their refrangibility very considerably. At 
 the same time, this increase makes it necessary to moderate the 
 light as much as possible, and then the feeble light will not allow 
 the higher eye-pieces to be used, without which the very minute 
 granules of myelin are not seen. Fuchsine, too, unfortunately is 
 apt to render somewhat indistinct the internal strise of the particles 
 and masses of myelin. 
 
 Aniline Blue. — This stain is inferior to the foregoing as regards 
 making out either the different forms of myelin and bacteria from 
 the fresh sputum, and so I have only used it experimentally. 
 
 Carmine. — This is most useful to determine the internal dispo- 
 sition of parts and the gortical layers of the vesicular gemmse of 
 the tuft, d. Fig. 3. As I observed in a preceding paragraph, it 
 does not stain any form of myelin, even when portions of sputum 
 are left in it for several weeks. I have always used Beale's carmine 
 stain. This solution, on account of the glycerine, renders almost 
 invisible the myelin granules, and even the particles and masses of 
 myelin are barely seen in it. This may be because the refraction 
 index of myelin and glycerine are about the same value, or at 
 least because the difference is too small for distinct vision.* 
 
 Resume. 
 
 From the foregoing remarks I may briefly conclude — 
 I. — In severe cases of whooping-cough, during the period of 
 convulsions, myelin is found in the sputum in the greatest abund- 
 ance, and bears no relation with the number of ellipsoidal epithe- 
 
 * The correspondence between the myelin found in sputa with the myehn 
 of the nerve-fibres should be noticed. In staining the nerve-fibres with carmine 
 the medullary sheath is not stained, while the cylinder axis is stained a full 
 red (Frey, Man di Teen. Micr., Naples, 1873, pp. 236 and 245. Ranvier, 
 loc. cit., pp. 97, 98 and pp. 722 — 724). I have recently tried all the above 
 methods of staining on myelin flowing from nerve-fibres and on preparations of 
 nerves from animals recently slaughtered, with the result that I find that myelin 
 in sputa behaves in exactly the same way as myelin of the nerves. This is sur- 
 prising, especially when picric acid is used, for a preparation of healthy sputum 
 mixed with myelin from nerve-fibres cannot be distinguished from the sputum 
 of whooping-cough. I hope to»treat this matter on another occasion. 
 
72 SPtTTA OF WHOOPING-COUGH. 
 
 Hum cells, nor with any inflammatory or febrile symptoms, but 
 solely with the intensity of nervous excitement. It certainly 
 cannot be imputed to any idiosyncrasy of individual patients. 
 
 II. — That though there is a formation and elimination, daily, 
 of small quantities of myelin from the air-passages in the state of 
 health, in no other state or affection of these passages is such a 
 remarkable quantity ever observed as in cases of whoopinE;-cough, 
 if we except the one case of haemoptysis mentioned by Beale; 
 
 III. — The extraordinary abundance of myelin may account for 
 the viscosity and transparency of the sputa of whooping-cough, 
 and also for the difficulty of expectoration, and may in many 
 cases serve as a guide in diagnosis. 
 
 IV.— That if the facts I have stated in this memoir are con- 
 firmed by further observations, and the. nervous phenomena of 
 whooping-cough are proved to be connected with the very great 
 abundance of myelin in the sputa in other epidemics and in other 
 countries ; this constant connection may open a new view of the 
 pathogenesis and etiology of whooping-cough, or may at least 
 induce others to undertake similar researches on the nature of 
 expired air, on the structure or condition of the medullary sheath 
 of those nerve-fibres that secrete myelin, and which are distributed 
 throughout the mucous membrane of the air-passages, and which 
 lie as a network just beneath the layer of epithelium at all points 
 where the mucous membrane extends. 
 
 V. — That the bacteria generally found in sputa, and particu- 
 larly in the sputa of whooping-cough, arise from germs, in all 
 probability, lodged in the nose or mouth ; so that, until the con- 
 trary is proved by definite facts, their presence in sputa should be 
 held to be in no way connected with the cause of the disease. 
 
 VI. — That the bacteria — though of many and divers forms — 
 can be reduced to two types, of which they seem to be only so 
 many different forms or phases of transition. One group can be 
 identified with Bacterium termo, the other with Leptothrix, and 
 probably these two belong in reality to one species. 
 
 WI. — That in any case, by simply relying upon the forms of the 
 bacteria found in sputa, it is impossible to judge with any certainty 
 at all whether the sputum examined be from a person in health or 
 disease, and on my second supposition, to determine the nature 
 
SPUTA OF WHOOPING-COUGH. 73 
 
 of the disease, except in cases of tuberculosis, in which Koch's 
 bacilli, chiefly of a cancellated appearance, are not bleached 
 under the usual test, and in those cases of pulmonitis and 
 pleuritis, where those apparently capsular forms are in the greatest 
 abundance, forms which nevertheless are met with in other sputa, 
 in acute bronchitis especially, they are found in large number 
 within the mucus corpuscles, and they are also found in urine and 
 the semen. But even in these excepted cases of tuberculosis, pul- 
 monitis, and pleuritis, the baderioscopical examination alone cannot 
 be assumed to be a positive and certain guide in diagnosis. 
 
 ^11^- — That any conclusion arrived at in a diagnosis from the 
 discovery of Koch's bacilli in phthisical sputum must, in many 
 cases, be doubtful. For even admitting that such a bacillus is the 
 cause of tuberculosis, which has yet to be proved, it has not, in 
 the first'place, any special morphological characteristics ; and, in 
 the second place, it is decolourised by the action of nitric acid and 
 alcohol, though perhaps in a less degree than are the other bacilli 
 and bacteria of sputa, if the decolourising test acts too far. 
 
 IX. — That fungi are found in whooping-cough sputa only in a 
 few cases ; judging by the germination of the species observed, 
 one appears to belong to the genus Mucor, and the other to the 
 genus Aspergillus ; in all probability both of these are inhaled 
 from the air. 
 
 X. — That gentian violet is to be preferred in the examination 
 of sputa in the fresh state, and this because of its double staining 
 - value ; the myelin appearing blue, and bacteria and other mor- 
 phological elements violet. The solution of iodine, picric acid, 
 and fuchsine come next in point of usefulness ; at the same time 
 carmine has its special use in showing the internal formation of 
 the vesicular gemmules of one of the fungi. 
 
 These conclusions are based on the examination of nineteen 
 specimens of sputa, taken in fifteen cases of whooping-cough, out 
 of which I have given particulars of nine. This is not a large 
 number, but these epidemics of whooping-cough are not frequent, 
 and I may have a long tin:\e to wait to renew my researches, and 
 gather still further information. I have therefore preferred, 
 without any delay, to bring what facts I have already learnt before 
 this august academy (R. Acad. Med.-Chir., Napoli), that others, 
 
74 SPUTA OF WHOOPING-COUGH. 
 
 when the opportunity offers, may be urged to verify the results of 
 my observations, and ascertain positively whether or no the 
 abundance of myelin be a constant feature of whooping-cough, or 
 be due to the peculiarity of a single epidemic, which by chance 
 has fallen under my notice.* 
 
 To any thoughtful mind the question must arise. How could 
 such an abundant flow of myelin, as I have described, fail to be 
 noticed by preceding observers? How is it that it has never 
 been mentioned by any of the many authors I have quoted ? I 
 have asked myself this question, and it seems to me that the only 
 answer is that either past examinations have not been fully carried 
 out, or that the presence of myelin has been treated as a fact of 
 Uttle importance, or else that casual phenomena has drawn me 
 into error. Finally, I should recommend that anyone who desires 
 to investigate into this matter should remember that myelin may 
 be scarce, or even non-existent in sputa during the first or 
 catarrhal period of the disease, and also during the close of the 
 attack. And to bear in mind, too, that the scarcity or absence of 
 myelin during the convulsive period is not an argument against 
 my hypothesis, viz., the lesion of the medullary sheath of the 
 nerve-fibres of the mucous membrane of the air passages. For 
 this may be affected, and yet myelin be kept from flowing into the 
 air passages whenever the minute bronchise are solely attacked by 
 the disease, or when, by chance, a portion of sputum be examined 
 coming from these smaller bronchiae. A final opinion cannot be 
 formed unless there is every certainty as regards the origin of the 
 sputum, and no doubt left that it did proceed from the throat 
 merely, or from the nose ; in such cases it is but natural that no 
 myelin would be found. There may even be cases where myelin 
 is totally absent, and this not because the nerve-fibres are intact, 
 but because the irritation has not reached the point of breaking 
 up the medullary sheath from which the myelin is given off. 
 
 * The present memoir had already been prepared when, at the end of 
 September and the beginning of October, there reappeared in this town several 
 cases of whooping-cough. I have, therefore, been able to compare my obser- 
 vations, and find that they entirely confirm my previous conclusions, particu- 
 larly with respect to the myelin and the fvingi ; should there at any time arise 
 any facts which may induce me to modify these conclusions, or merit any 
 especial consideration whatever, I shall be pleased to bring the same before 
 the notice of the academy. 
 
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[ 75 ] 
 
 IRecent Bacterlologfcal IResearcbes on tbe Sputa ; 
 tbe /iDorpbologs anb BIoIoqs of tbe /lOfcrobes 
 of tbe /iDoutb. 
 
 By Filandro Vicentini, M.D., Chieti, taly. 
 Translated by Professor E. Saieghi. 
 
 Further Remarks on the Bacteria and Bacilli found 
 IN THE Sputa, 
 
 and their relation to the " Lepfothrix bu'aalis ;" the result of 
 original observations on the morphology and biology of this 
 parasite, principally on its higher phases, now for the first 
 time examined and described. 
 
 (Continuation of the Memoir on the Sputa of Whooping-Cough, Vol. XLIII., 
 of theJ'Atii Accademia, Medico-Chirurgica di Napoli") 
 
 " The manner of reproduction and the reproductive bodies of the Lepiothrix 
 are not known ; but, perhaps, more accurate research will discover them : 
 this hypothesis being based on the fact that, in the interior of filaments 
 magnified to 800 diameters, small, round bodies are seen which might be 
 spores." — Robin. 
 
 Summary : 
 
 § I. — Further remarks on bacteria and bacilli found in the sputa 
 and contents of the mouth — A rhum& of the preceding 
 work — Some forms of bacteria not therein described — Other 
 forms of bacilli, ut supra. 
 
 § 2. — Summary of the present investigations and methods of tech- 
 nique — Collecting and preparing the sputa and the buccal 
 contents. 
 
 § 3. — Previous opinions on the micro-organisms of the buccal 
 cavity, according to Miller — History and general facts — 
 Lepiothrix buccalis (Lepiothrix innominaia, M.) — Other 
 buccal micro-organisms — Buccal pathogenic fungi. 
 
76 BACTERIA IN THE SPUTA 
 
 § 4.— Observations on the morphology and biology of Leptothrix 
 buccalis—The. four phases of Leptothrix, particularly the fruc- 
 tification by spores— Production by points (male organs, ?) 
 — Fructification of spores reproduced in the sputa — Dissemi- 
 nation of microbes in the posterior organs. 
 
 Recapitulation. Bibliography. 
 
 §1- 
 
 FURTHER REMARKS ON THE BACTERIA AND BACILLI 
 
 FOUND IN THE SPUTA AND IN THE CONTENTS 
 
 OF THE MOUTH. 
 
 AT the meeting of 2Sth November, 18S8, I had the honour of 
 presenting to this illustrious Academy a memoir on the 
 sputa of Pertussis, in which I incidentally touched upon 
 sputa of another character. 
 
 It is not my intention to re-open all the questions before treated, 
 but simply to refer to the bacteriological aspects. The recent 
 observations refer to all bacteria and bacilli found in normal sputa 
 as well as in those of various morbid conditions, the so-called 
 tubercular bacilli not excluded, it being understood that the 
 many special questions belonging to the latter bacilli will be 
 fully considered later on. 
 
 Confining my remarks to the bacteriological researches in 
 sputa, I will briefly repeat the conclusions I came to in the 
 preceding memoir : — " The bacteria generally found in the sputa, 
 and particularly in those of Pertussis, or Whooping-Cough, are 
 derived, in all probability, from germs originating in the mouth 
 and nose, so that (and until irrefutable proof to the contrary) their 
 presence is to be taken as a natural fact, which has nothing to do 
 with the cause of the disease. Such bacteria, although of very 
 varied forms, can be reduced to two sole types, representing as many 
 phases or forms of transition. One group may be identified with 
 Bacterium iermo and the other with Leptothrix; and probably even 
 these two groups belong to a single species." 
 
AND CONTENTS OF THE MOUTH. 77 
 
 And elsewhere :— " In summing up the feature of the different 
 types, we may conclude that the many forms of Bacteria delineated 
 in our plate, together with the others simply pointed out in the 
 text, can be reduced to two species, viz. : Bacterium termo and 
 Leptothrix. 
 
 The forms a, b, c, above (Fig. 2, PI. I.), d, e,f,f, g, i, k, I, t, 
 p, and p', apply to Bacterium termo ; the forms V, c (below), m, n, 
 «', (left), q, r, s, u, v, if, x, x', y, and z, to Leptothrix. And it is 
 not improbable that, in their turn, the two species, according to 
 Cohn, Naegeli, De Bary, and others, may form only one. These 
 authors classified the round Bacterium (monas crepusculum). Bade" 
 rium termo and Leptothrix, as three forms or developments of one 
 and the same species. 
 
 I am rather inclined to believe that even the forms known 
 under the names of Vibrio rugula, clostrium, and rhabdomonas may 
 be attributed to the same species, as three simple varieties of its 
 bacillary forms." I base these conclusions upon indirect signs, 
 and especially upon certain morphological characters, common or 
 transitional, on the usual habitat, etc. 
 
 Now, having followed up this line of research for the last two 
 years, I think I may be able to give a direct demonstration of my 
 work, by indicating, so to speak, the explanation of the polymor- 
 phism of the various microbes lodged in the sputa. 
 
 Moreover, in the preceding paper, several types of Bacteria 
 and Bacilli, for brevity's sake, were overlooked, as well as other 
 specimens drawn from further observations.* " 
 
 * Whilst again touching on Whooping-cough, and in continuation of our 
 bibliographical notices, we may mention a late article from Mircoh on " Renal 
 Alterations in Pertussis " (Arch. p. le Sc. Meduhe, 1890, p. 63, etc.) Mircoli 
 inoculated, ineffectually, the cultures of bacteria, taken from the larynx of 
 children who had died from Whooping-cough, into the larynx or under the skin 
 of rabbits. He mentions articles from Sseurtschenho and Wendt, confirming 
 the observations of Afanasieff. In the Journal of Microscopy and Natural 
 Science, edited by Alfred Allen (January, 1890). there is a Lecture given by 
 Dr. Shingleton Smith before the Microscopical Society of Bristol ( "On Some 
 Recent Developments of the doctrine of a Contagium Vivum " ), where (at p. 
 32) it is said that the efficacy of special micro-organisms in most of the con- 
 tagious diseases, and particularly in those most common to children, as 
 Measles, Whooping-cough, and Scarlet fever, is not proved. 
 
78 BACTERIA IN THE SPUTA 
 
 Some forms of Bacteria not described in the preceding 
 Memoir (Fig. 4). 
 
 All forms of Bacteria and Bacilli herein described (Figs. 4 and 
 S) are coloured with gentian violet, excluding the filament or ser- 
 pentine bacillus, h! (Fig. 5), which has been stained with iodine 
 solution. 
 
 {a) In the preceding Memoir I maintained that the typical 
 curved Diplococci— ^, /, p (Fig. 2) — which are morphologically 
 identical with the Gonococci of Neisser, never present a capsular 
 appearance, and this proves to be so in most cases. But from 
 further observations I have found exceptions to this rule, as is 
 shown in Fig. 4« (magnified to 690 diameters). Here, in fact, the 
 curved Diplococci are intermixed with straight Bacteria, on parallel 
 cross lines, and are enclosed with the same envelope, which con- 
 tains the whole. I found on December, 1890, similar specimens 
 for the first time in the thick, opaque sputa of a child of seventeen 
 months affected with Whooping-cough, and on the seventh day after 
 the expectoration. The sputa contained, in the midst of common 
 bacteria and bacilli, a fair number of red blood-corpuscles with a 
 few ellipsoidal epithelia and granules of myelin. But, later' on, I 
 accidentally met with similar specimens in urine. This is no cause 
 for surprise, as I believe that the impregnation of microbes in the 
 urinary and spermatic passages to be the same as that of microbes 
 vegetating in sputa, as they proceed from the same parasite, Lep- 
 totkrix, which vegetates both in the mouth and in the external 
 genito-urinary passages, as I shall prove later on. 
 
 In the third section I hope to show likewise the analogy between 
 this type of Bacterium and the Jodococcus vaginatus of Miller. 
 
 But, besides these curved diplococci, which are intermixed with 
 straight bacteria in a common envelope, others are to be found 
 coupled only and enclosed in one sheath by pairs (seldom) in the 
 patina dentaria (white deposit upon the teeth) around the rich 
 colonies of Diplococci. Their capsular appearance is made 
 striking, owing to the staining with picric acid and successive 
 saturation in glycerine. 
 
 <5, v. — This specimen (magnified to 1750 diameters) is taken 
 from \he patina dentaria, mixed with saliva ; groups of diplococci 
 are seen in it, both large and of medium size, interwoven with 
 
AND CONTENTS OF THE MOUTH. 79 
 
 Strings of very minute cocci, morphologically identical with those 
 of type s (Fig. 2). 
 
 Such specimens, as well as diplococci with the capsular appear- 
 ance of type / (same figure), reproduced in d' (Fig. 4), are num- 
 berless. These diplococci are of two sizes, large or medium of 
 type I' (Fig. 2) and others, surrounded likewise by a clear sheath, 
 but small, as in d' (Fig. 4, right). The small cocci enclosed in i' do 
 not differ materially from those of the beaded forms (with the 
 exception of being surrounded by a sheath). I have observed that 
 these minute diplococci are the heavier, because they are found 
 mostly on the slide, whilst the large diplococci adhere to the 
 cover-glass. 
 
 Now, the specimen drawn in & shows that the beaded forms, 
 morphologically identical with the bacilli of Koch, are found in 
 the patina dentaria, as well as in the solid and stagnant substrata, 
 viz., within the buccal epithelia or in the morbid secretions, either 
 in the crypts of the larynx or the tubercular products, in so far as 
 the inactivity of the part favours the multiplication of the lineal 
 series of cocci, which first form strings of beads and then small 
 rods or filaments, as I have already demonstrated. 
 
 In the second place, from this specimen, I believe there exists 
 a close afJSnity between those beads and diplococci, either of the 
 buccal cavity or of some other habitat ; otherwise I cannot under- 
 stand why they should be so close together as to resemble fruits 
 fallen from the same tree. 
 
 Another example of the tendency those minute cocci have to 
 adhere to the diplococci is seen in /. 
 
 (c) The couples of dumb-bell bacteria herein shown (magnified 
 to 2,500 diameters) are taken from the nasal mucus in its normal 
 condition. I contend that they do not differ at all from the ordi- 
 nary dumb-bell and chain-like bacteria found in the sputa upon 
 the types c, d, e (Fig. 2), which remind us of the form and size of 
 Bacterium termo. 
 
 On examining thick nasal mucus, I met with myriads of such 
 bacteria. In this case the material to be examined must be taken 
 from the dry cavities of the nose by a glass rod. 
 
 We shall see that those dumb-bell bacteria are more apt to form 
 chains ; in fact, they constitute the remarkable groups or bundles 
 of chain-like bacteria found on the surface of the tongue. 
 
80 BACTERIA IN THE SPUTA 
 
 According to my researches, each dumb-bell is the result of 
 two cocci linked together by their heads. For this reason, it is 
 often found, enclosed by a sheath, either inside or outside of the 
 epithelia of the mouth, nose, or urethra. It is this adhesive ten- 
 dency which forms them into chains or groups, from which in less 
 moist parts will germinate the growth of Lepiothrix, as we shall 
 see later on. 
 
 I must refer to the preceding Memoir for the other forms of 
 bacilli and bacteria, all related to Lepiothrix. With regard to the 
 nasal mucus, its pavement epithelia are smaller than the same epi- 
 thelia of the mouth and less softened, perhaps from not being 
 under the dissolving action of the saliva. 
 
 The bacteria which invade the pavement epithelia of the 
 urethral mucus, taken directly from the meatus, often resemble 
 dumb-bell forms. However, in the blenorrhcegic flow, are often 
 found rather small diplococci, some in the pavement epithelia and 
 others within pus corpuscles ; but curved diplococci or gonococci 
 of Neisser are rarely found in it. Beside these common forms, I 
 also detected very minute cocci, like those rosaries of type s (Fig. 
 2) or ^ (Fig- 4)1 ^nd filaments or short articulations of Lepiothrix. 
 
 d, e. — I have said before that the features of pneumococci of 
 Friedlander, or Frankel, etc., were probably derived from a 
 degree of colouration of some points of their single articulations 
 or from modifications of forms, as types d and e will Show. 
 
 In d (Fig. 4, magnified to 2,500 diameters) are seen three 
 sheathed diplococci, partially coloured. 
 
 In certain cases of pneumonia, we found some specimens of 
 these, intermixed in the same group or colony. 
 
 In e the diplococci have no sheath and show points of varied 
 partial colouring. Such specimens were obtained from a pulmon- 
 ary sputa, in which, amongst other forms of bacilli and bacteria, 
 were pneumococci of types j'j g, h, i, i'. 
 
 /) /'i i- — These are specimens of the second kind, taken from 
 the same sputum in a case of croupal pneumonia. In / (magni- 
 fied to 1,750 diameters) the capsular appearances are so well 
 surrounded as to resemble a capsular membrane. The single 
 pneumococci are sometimes in groups of eight, ten, and fourteen. 
 The internal articulations are prolonged and end in points. 
 
AND CONTENTS OF THE MOUTH. 81 
 
 In/' (magnified to the same degree) the articulations are also 
 acuminated, but have no halo, and at first are very pale. In suc- 
 cessive days they become partially coloured in the preparation if 
 kept artificially moist. These pale Diplococci, without sheath, 
 appeared to be in a state of active germination, as we detected 
 therein the more minute and proportionally paler forms, some 
 being composed of unequal cocci, and several were moving about 
 in the medium. The forms surrounded by sheath are firmer or 
 less active, as if the capsule indicated the quiescent state of the 
 microbe. In fact, if it moves, it is by a motion of simple transla- 
 tion, without vibrating, as is the case with bacteria in active 
 germination,* 
 
 It appears that the capsule, owing to its great refraction, and 
 also to the difficulty the near small bodies find in crossing it, 
 resembles a fatty substance surrounding the microbe or proceed- 
 ing from it. This is only an hypothesis. 
 
 In g (magnified to 2,500 diameters) is shown a distinct diplo- 
 coccus, but with the internal articulations nearly cylindrical, with a 
 large and somewhat irregular capsule, resembling the mono-articu- 
 lated bacillus of type q (Fig. 2). The sheath indicates, even here, 
 a quiescent state of the microbe and the successive secretion of 
 pale substance. Some analogy may be found in it to the Bacillus 
 crassus sputigenus of Kreibhom. (See section 3.) 
 
 h. — Intermixed with the diplococci e and /' we find, at times, 
 in the pulmonitic sputum, these forms without a sheath (magnified 
 to 2,500 diameters). In one of these forms we infer the process 
 of increase by seeing the pointed articulation at the right part as 
 pale as in /'. The lengthened part of the mono-articulated 
 bacillus is strongly coloured, and, in a good light, exhibits internal 
 punctuations of deeper colour, especially near the edges : a remark- 
 able circumstance that makes them resemble the Leptothrix, in 
 which are also these punctuations or future gemmules. 
 
 The above forms are met with in the second period of pul- 
 monary affections after the fever has subsided, but may also be 
 
 * In a recent work, Billet states he has met in other species with this form 
 of incapsulated diplococci, and considers it a phase of life common to various 
 species of bacteria. He calls it the zoogloeic state. (This will be again 
 referred to in a note at the end of Section 3.) 
 
82 BACTERIA IN THE SPUTA 
 
 found in successive periods. These pneumococci retain tena- 
 ciously the gentian violet, as I stated in the first memoir. 
 
 Several of the preceding forms of pneumococci, particularly 
 those of types d, e, f, /', and h, are analagous with those drawn by 
 Cornil and Babes in Les Bactkries etleurrdle, PI. XV., Figs, i, 4, 
 6, and 13.* 
 
 i, i'. — The large round diptococcus, i (magnified to 2,500 
 diameters), was also found in the second period of a pulmonary 
 affection. Its articulations resemble two detached hemispheres, 
 and round the envelope adhere very minute cocci, not one of them 
 being able to penetrate the clear zone. This fact clearly indicates 
 the solidity of the sheath. 
 
 In /' there is a small chain-like bacterium, the articulations of 
 which exhibit a series of similar diplococci, so connected that the 
 hemisphere of the one adheres to that of the other, leaving a 
 clear space between the hemispheres, of each diplococcus. Such 
 chain-like bacteria are sometimes found grouped together, although 
 in small number, in pneumonic sputa. 
 
 Other Forms of Bacilli, ut supra (Fig. 5). 
 
 Some forms of bacilli were omitted in the other memoir for 
 want of space ; others were collected later on, and these I will 
 now briefly describe. The new forms added to the first will better 
 show the identity of the bacilli of the sputa with the articulations 
 or fragments of the filaments of Leptothrix vegetating in the mouth. 
 
 In the preceding Memoir the bacillus n' (Fig. 2) represented a 
 form of transition from the young bacillary forms, n, n, n, to the 
 mature forms, m, m. I then pointed out other forms of transition. 
 
 a, b, c. — In a (Fig. 5, magnified to 1,250 diameters) is seen a 
 bacillus that partakes of three distinct types, divided into three 
 segments : one internally occupied by four ellipsoidal bacteria, the 
 clear segment in the middle, slightly coloured, and the other 
 deeply coloured and opaque. It shows that the clear bacilli 
 inside — those containing bacteria and those entirely opaque — 
 belong to the same species. This bacillus was taken from influ- 
 enza sputum, but similar forms were found in other sputa and in 
 the saliva. 
 
 * Work quoted in the Bibliographical Appendix. 
 
AND CONTENTS OF THE MOUTH. 83 
 
 Another analogical proof we have in b, where a bacillus, opaque 
 for five-sixths of its length, exhibits the other one-sixth clear and 
 slightly coloured. This specimen was taken from the saliva. 
 
 In c we find four types — viz. : a short, clear segment above, 
 and on the top of it a bacterium of elliptical form ; in the middle, 
 a long, opaque segment ; and in the lower part a pale, granular 
 segment like the middle feature of type n' (Fig. 2) and types e and 
 e' (Fig. 5). This specimen was also taken from the saliva. 
 
 d. — This short but large bacillus, taken from the patina den- 
 taria (magnified to 2,500 diameters), exhibits the upper part 
 opaque and deeply coloured ; the other part is divided into two 
 segments. The middle segment is very short, clear, and contains 
 a dumb-bell bacterium placed across ; the lower one is opaque, 
 but not entirely, as there is on the right a clear space between the 
 opaque part and the outline, and presents a kind of indentation.- 
 Other bacilli have the same features towards the edges, or near the 
 junctions of. their segments. 
 
 As regards the enclosed bacterium, there is no doubt about its 
 position in the envelope of the bacillus, from the look of the spe- 
 cimen as well as from the oscillation of the preparation, in which 
 the bacterium was seen to be always inside. In the preceding 
 specimens we have also abundant proof of this fact. 
 
 Now in the first place, it shows that not only elliptical, very 
 minute, and linear bacteria, but also dumb-bell bacteria, may be 
 found in the envelope of the bacilli ; from which we conclude 
 there is no difference between the dumb-bell bacteria and the 
 bacilli. And since the small chains, and the bundles especially, 
 adhering to the tongue, are mostly formed of dumb-bell bacteria 
 we must infer the analogy of those forms with the bacilli, and 
 consequently of both to the Leptothrix. 
 
 In the second place, comparing the various specimens of bacilli 
 (as a and d), containing bacteria with the fertile filaments of Lep- 
 tothrix, we find in the latter a variable disposition of internal 
 granules, or buds, adhesive to the envelope. Some are alternate ; 
 others on the same level at both sides. If we imagine that in 
 those filaments the buds grow up towards the middle line of the 
 stem, it will result that, when the left bud is placed higher than 
 that on the right, the bacteria resulting from their increase will 
 
84 BACTERIA IN THE SPUTA 
 
 range themselves in a longitudinal line, taking the elliptical form, 
 as in bacillus a. On the contrary, if the two buds are on the same 
 level, we shall have the case of bacillus <^— viz., the two cocci, 
 resulting from the increase of the buds, will meet in the centre of 
 the stem, and, there joining together, will produce, in a cross line, 
 the dumb-bell bacterium. 
 
 Therefore, wherever we turn our eyes, the notion of the poly- 
 morphism of the microbes, and their probable derivation from a 
 single species, is gaining firmer ground. 
 
 e, /.—The bacillus drawn in e (Fig. 5, magnified to 1,750 diam.) 
 was found in the sputa of a young woman in labour, who was 
 affected with bronchitis. It was mixed with other similar granular 
 bacilli, clear and of the same length, but more slender, as we see 
 in / These bacilli, however, are common also in other sputa and 
 in the contents of the mouth; they present a pale, granulated 
 mass, similar to that of bacilli, n' (Fig. 2) and c (Fig. 5). They 
 also show, in various places, buds or internal granules, smaller and 
 paler than those found in the fertile filaments of Leptothrix. 
 
 In e the ends are pointed, but in / they are round. The larger 
 of "the two is faintly coloured, but exhibits near one of the heads a 
 clear interstice, as is seen in a and d. The other specimen, 
 smaller and paler, exhibits a younger form of the same type. 
 These two last bacilli were found in pulmonitic sputa, and were 
 also found in the contents of the mouth. 
 
 In the uncoloured preparations of the patina dentaria there are 
 nail-shaped bacilli, like the type e, soft, flexible, slightly veined, 
 and having quick motion (Fig'. 14, d). These bacilli are originally 
 lodged on some pointed productions, as was observfed by me for 
 the first time ; their slender points form the bacilli /i h, and h'. 
 
 /, g, h, H. — The very slender bacilli, / and g (magnified to 
 2,500 diameters) are also taken from a case of pneumonia. In 
 / we see a mono-articulated and strongly coloured bacillus, as well 
 as another pale and granulated. In g, a very slender one, the 
 ends of which appear to have been recently detached from a 
 longer filament. 
 
 By comparing these specimens with the Leptothrix, no import- 
 ant difference is noticed. 
 
 In h another specimen of curved bacillus, resembling a point 
 
AND CONTENTS OF THE MOUTH. 85 
 
 of interrogation, taken from pulmonitic sputum. These bacilli are 
 not uncommon in the patina deniaria. They have quick vital 
 movements. They resemble the Spirillum sputigenum of Miller 
 or the Comma bacillus of the saliva described by Lewis, as 
 identical with the cholera bacillus of Koch, Bacillus virgola.* 
 
 I have, however, found that, between these bacilli and the 
 lesser points, which are abundant on certain filaments of Lep- 
 tothrix, there exists a great similarity. These points vary in size, 
 but all possess quick movements (Fig. 14). We shall consider 
 later on if these bacilli act like the reproductory filaments (sper- 
 matia or antherozoids) of many cryptogams, although in Leptothrix 
 they are not originally contained in special receptacles (spermo- 
 gones,or antherids). 
 
 In a more advanced hypothesis, we shall have also to consider 
 if these bacilli represent the spirilli, or if the spirilli proceed from 
 such filaments, endowed with the same motile power. It appears 
 more probable that the spirilli may generally be derived from the 
 fragments of certain curved filaments, fertile or not, of Leptothrix. 
 Sometimes the spirillum is thicker than the filaments of Leptothrix, 
 as in Fig. i of Cornil and Babes.t 
 
 In h' is seen a larger filament (stained with iodine), found in 
 ihs patina dentaria, which is somewhat similar to spermatozoa. 
 
 In the uncoloured preparations I have seen the very rapid 
 movements of all these bacilli. In conclusion, all these forms of 
 bacilli are common in the expectorations and contents of the 
 mouth. 
 
 § II. 
 
 SUMMAEY OF THE PRESENT INVESTIGATIONS AND 
 
 METHODS OF WORKING. 
 
 Argument. 
 In the fourth section of my Memoir on Whooping-cough, I 
 stated that bacteriological researches on the sputa would be fruit- 
 
 * Lewis, A Memorandum on the Comma-shaped Bacillus, etc. 
 
 t Cornil and Babes, op. «V.— Vicentini, "Sopra un caso di febbre ricorrente ' 
 ecc. e sul riscontro degli Spirilli nA Sungue," Jtti del. R. Accad. Med. Chir. 
 di Napoli, 1883, t. xxxvi. 
 
86 BACTERIA IN THE SPUTA 
 
 less until the natural history of the pathogenic or non-pathogenic 
 microbes that lodge there is thoroughly known. 
 
 In most cases a complete organism grows in length, breadth, 
 and thickness. But the chain-like bacteria and the filaments only 
 grow in one direction (in length) simply by a repetition of particles 
 in a lineal series. We must, therefore, suppose that neither these 
 original particles, nor the chain-like bacteria and filaments resulting 
 from them, are complete organisms, but that they are rather rudi- 
 ments of more perfect ones. 
 
 By examining all the different forms of bacteria and bacilli, we 
 were led to believe in a single type, to which all other forms were 
 related. Now, we will show that the existence of this vegetation 
 is to be found in the patina dentaria, in certain sputa, and jn the 
 mucus of the urethra, all the forms hitherto described being only 
 fragments of it. 
 
 If somebody, for instance, not knowing all the phanerogams, 
 should enter a wood-house, looking at the wood, heaps of branches, 
 leaves, etc., he might at first suppose that they were different 
 things instead of parts of a whole. Likewise, if he should see a 
 threshing-floor upon which is spread and beaten Indian corn, he 
 might take the stems, roots, leaves, tufts, and grains for as many 
 different objects. But it will be sufficient to take him out into 
 the field, to convince him of his mistake. Looking through 
 the lines of the corn, he would see that most of the plants (the 
 female ones) are provided with ears, and he would see also others 
 higher than the rest (the male ones), having instead of ears only 
 a tuft. 
 
 Then he might suppose that the plants of this second type 
 were, perhaps, of different kind from the others, not thinking that 
 Indian corn is a dioecious plant, with distinct sexes on different 
 individuals. 
 
 This is precisely the case with the microbes of sputa. We 
 were led to the study of the morphology and biology of Leptothrix, 
 by considering that nothing is useless in nature ; those parts which 
 now appear superfluous to us in living organisms, (as the nipples 
 in man) are rare exceptions. 
 
 The illustrations of Leptothrix contradict this law. Look, for 
 instance, in Bizzozero, the figure of Leptothrix buccalis, reproduced 
 
AND CONTENTS OF THE MOUTH. 87 
 
 in our Fig. 6, and the query will be asked :— Of what use are those 
 upright, barren shoots resembling grass, which have been eaten 
 close by cattle ? Cannot this want of fruits be accounted for by 
 mechanical injuries? The figures of Robin, Frey, Miller, and 
 others do not suggest a different conclusion.* 
 
 After these reflections, we were induced to investigate this 
 parasite. Our researches will be particularly demonstrated in 
 Section 4, only a synthesis of them being given here. 
 
 From our observations Leptothrix buccalis would, therefore, 
 live under four forms, or have four different stages of development, 
 according to the conditions of the alimentary substratum. 
 
 In the liquid secretions, as saliva and mucus, its spores— bac- 
 teria and cocci — and its gemmiferous sprouts — real bacilli (exclud- 
 ing the Spirilla, the nail-like, the snake-like, and the comma) — 
 increase and multiply like moving spores and germs of fungi. 
 This would be the immersed vegetation of Leptothrix. 
 
 The bacteria, the yeasts, and the spores have generally the 
 power of reproducing the full organism like the seeds of the 
 phanerogams ; but, between the spores of the bacteria on one 
 side, and the seeds of the phanerogams on the other, there is this 
 difference : that those seeds cannot multiply by division, produc- 
 ing fresh seeds ; whilst moving spores, yeasts, and bacteria multi- 
 ply abundantly, by fission, even by themselves. 
 
 If, however, the liquid is kept quiescent in the crypts or cavi- 
 ties, or if the part is, as on the surface of the tongue, very firm, 
 the second period or low vegetation begins, viz., beaded, chain-like, 
 and bundle-like bacteria are formed, so common on the patina of 
 the tongue. But vegetation stops here, for in the mucous parts 
 the continual friction impedes a further formation. 
 
 In the secretions and mucus there is a relative inactivity ; but 
 if it becomes effective, a reproductive growth will be formed, as 
 shown in Fig. 16, taken from pulmonitic sputum, which for 
 several days had adhered in the interior of a closed tube. 
 
 The third period, or growth, is when it lodges on a solid part, 
 
 * Bizzozero, Manuale cit. nella Bibliografia, PI. II., Fig. 27 ; Robin, His- 
 toire naiurelle des Vegitaitx, etc., 1853 (see Bibliography), PI. I., Figs> i and 2. 
 Frey, figure reproduced by Perroncito, quoted in Bibl., Fig. II. Miller, 
 figures quoted later on. 
 
88 BACTERIA IN THE SPUTA 
 
 on the tongue or teeth. But, owing to the continual friction on 
 the tongue and teeth, the growth cannot reach the fourth period, 
 or that of fructification. Then we have bare growths apparently 
 sterile, as represented above. The same happens with the tufts of 
 Leptothrix, often found on the epithelium scales, on the residue of 
 undigested food, or other corpuscles in urine, sputa, etc., forms 
 which may be called " aerial," although incomplete, " vegetation." 
 
 But in many parts of the dental surface, where there is less 
 friction, as between tooth and tooth and near the gums, the fourth 
 phase or fructification takes place. This fact has not been ob- 
 served, and is not even suspected, as we gather from the most 
 recent work of Miller.* 
 
 I call it aerial vegetation, because the stems of the microphite 
 have a tendency to rise in the air, although immersed like 
 the algae. In our case the surrounding liquid is the saliva. This 
 fructification by spores, first noticed and investigated by me, con- 
 sists of comparatively long ears (Figs. lo, ii, 12, 13, 16), 
 formed by the exudation of a viscous substance, faintly coloured, 
 round the ends of fertile filaments. In this substance are entan- 
 gled small round spores, often brightly coloured, placed in six 
 longitudinal lines, as we shall see later. In the strongest speci- 
 mens (Fig. 16,/, g) as many as 720 spores can be counted in an 
 ear. All the cocci and minute bacteria are probably dissemina- 
 tions from these spores. 
 
 There is another form of special production, or pseudo-inflo- , 
 rescence, which is drawn in Fig. 14, a, b, c, resembling points, 
 round a filament or group of fertile filaments. These points 
 having fallen off form the nail-like, snake-like, or comma bacilli. 
 If we admit the analogy of these bacilli with the reproductive 
 filaments of other cryptogams, Leptothrix may be compared to 
 a fungus, or dioecious alga, with two sexes upon different fila- 
 ments ; and such pointed productions would exhibit the male 
 organs, the elements of which, spread about the surrounding 
 medium, would, with their great mobility, act as antherozoids in a 
 manner not yet known to us. In such an hypothesis, the 
 scanty number of such inflorescences, with regard to fructifica- 
 tion or female organs, would be fully explained. Therefore, 
 
 *W. D. Miller, Die Mikro-Organismen der Mundhohle, etc., 1889. 
 
AND CONTENTS OP THE MODTH. 89 
 
 in summing up the morphological and biological series of 
 Leptothrix, beginning with the forms of Cocci or Bacteria, we 
 pass through the chains, the growths, and end in the fructification 
 by spores, in order to begin again another series usque ad infini- 
 tum. But, beside this cycle of reproduction, we must notice 
 another — of buds and internal impregnation (that already foreseen 
 by Robin), which is seen in the interior of the envelope of old fila- 
 ments, articulations, and bacilli, and through which, from the 
 same bacilli, are generated other bacteria, and this owing to the 
 reserve buds described above. From those bacteria are, in their 
 turn, reproduced the chains, the tendrils, and their articulations or 
 bacilli, as we have seen in the former Memoir on the subject of 
 bacilli b' and n (below) and in u, u (Fig. 2). 
 
 Let us consider what a great cause of dissemination this is, 
 • placed at, the entrance of the digestive and respiratory organs. 
 According to our calculations, there would always be found present 
 in the nose and mouth, taken together, from two hundred to three 
 hundred trillions of bacteria, or other elements of the microphite, 
 ready to enter the stomach at the time of deglutition, and to fall 
 on to the respiratory passages at every breath. In comparison 
 with that mass, what are the few germs we inhale from the 
 atmosphere ? 
 
 But if, at every breath we inhale, a propelling motion sends 
 whole swarms of elements or germs of the microphite into the air- 
 passages, another slower but continuous movement pushes germs 
 or analogous elements on into the genito-urinary passages 
 through contiguity, from one epithelium to another, and onwards 
 along the mucous patina, passing through the urethral orifice, 
 which Pasteur has compared, in this case, to the Thames Tunnel.* 
 
 We will briefly treat these questions in Section 4, and demon- 
 strate the identity of Leptothrix prepuiialis with Leptothrix buccalis. 
 Now, to enable others who may be willing to repeat and to verify 
 our observations, we will proceed to describe the modus operandi. 
 
 * Schutzenberger Le fermentazioni, ecc. trad., Milano, 1876, p. 204 ; V. 
 also Vicentini Caso di vegetazione di funghi microscopici nell' Uretra 
 ecc, Morgagni, April, 1 880. 
 
90 BACTERIA IN THE SPUTA 
 
 Collecting and the Preparation of the Sputa. 
 
 In the preceding work we dealt with the collecting and the 
 treatment of sputa, putting off to another occasion the diagnosis 
 of derivation of the various materials expectorated, in order that 
 the real sputa might be distinguished from the particles of food ; 
 the sputa from the saliva or throat, and the mucus flowing from 
 the nose, being also differentiated. Now is the moment to touch 
 on this point again, in order to describe improved methods of 
 preparation and colouring. 
 
 The materials to be submitted to microscopic observation 
 really do not always proceed from the 'air-passages. In the case 
 of a boy, 7 years old, affected with Whooping-cough, we remember 
 having received a portion of food ejected from the stomach, and, 
 thought to be a sputum. This substance was gelatinous in appear- 
 ance, the colour of tobacco, and contained a large amount of 
 aljmentary residue ; but what surprised us was a very dense 
 mycelium of conspicuous sprouts after the type drawn in former 
 plate, Fig. 3, d, interwoven with other slender ones. The fructifica- 
 tion of these mycelia produced forms identical with small heads 
 (capitula) or sporangia, delineated in k and ^', very vigorous, upon 
 as many as five fertile branches, intermixed with similar vigorous 
 forms of Aspergillus glaucus. This would lead us to suppose that 
 the deglutition of the relative germs with the sputa, and their 
 secondary development, is even more easy and vigorous in the 
 stomach. 
 
 In some cases, especially with children, the. proper expector- 
 ation is lost or swallowed ; and when the patient is asked to 
 spit out again, he emits only a httle saliva, mixed with small flakes 
 of mucus, from the throat and tonsils. These salivary sputa, how- 
 ever, may be recognised generally on the following day through 
 the clarification, after remaining undisturbed, with an upper liquid 
 stratum (more considerable than in crude sputa, or those mixed 
 with abundant saUva), and a scanty deposit, very rich in buccal 
 epitheUa, and, eventually, alimentary residue. Besides, they are 
 recognised from being wanting in ellipsoidal epithelia and granules 
 of myelin. 
 
AND CONTENTS OF THE MOUTH. 91 
 
 Another source of error, against which it is difficult to guard, 
 is the reflux of nasal mucus, through the posterior nares, in the 
 back of the mouth, and its successive expectoration mixed with 
 saliva. Naturally, ellipsoidal epithelia and myelin, even in such 
 sputa, are wanting. 
 
 In any case the presence of the myeUn and the ellipsoidal 
 epithelia induces us to regard the sputum as proceeding from the 
 air-passages. These are two important points when, for instance, 
 a doubt arises whether a sputum with a rusty appearance proceeds 
 from the nose or the chest ; as, in our daily researches on sputa, 
 we never detected ellipsoidal epithelia or myelin in spuripug sputa j 
 whilst in the genuine expectorations they are very seldom absent, 
 and when not found in the first, they appear in the second 
 preparations. 
 
 We come now to the manner of collecting the sputa- 
 
 The act of spitting is accomplished in two different efforts : 
 by the first, the genuine sputum, nearly entirely frel froro saliva, 
 and that which is required is coughed up ; but soon after by a 
 second effort, saliva is emitted with the residue of the sputum left 
 behind. This second sputum (being nearly all saliva) is elimin- 
 ated. In order to avoid mixing the sputum with saliva, the patient 
 should not keep it in the mouth too long. 
 
 Instead of a spittoon, it is better to use a colourless plate or 
 saucer, perfectly clean, where the sputa can be kept separate, and 
 then selected for examination. The sputum should be takep at 
 the most viscid point with a bent needle or forceps, and raised so 
 that the saUva and the more fluid part of the mucus with whiph 
 it is mixed, may trickle down on the plate. What remains on the 
 instrument is quite sufficient for successive investigatipns, an4 it will 
 necessarily contain very little saliva. To preserve it, it should at 
 once be placed in a glass tube, which has previously been cleaned 
 with sulphuric acid, and washed out with alcohol, 
 
 For the coloured preparations we have to modify the directions 
 given in the preceding work. It is better to keep the solution of 
 gentian violet separated from the aniline water, so as] to prevent 
 decomposition ; but we must avoid putting the glas? rod, still wet 
 with aniline water, into the colouring solution, and vicf versa. 
 Two different rods must be used. With a small rod put first on 
 
92 BACTERIA IN THE SPUTA 
 
 the slide a drop of aniline water, taken from the relative bottl , 
 and soon after add a colouring solution. For, by placing the 
 solution first and the aniline water afterwards, we obtain a greater 
 precipitation of coloured granules in the preparation. 
 
 It is necessary that the particle of sputum should be well 
 immersed in the colouring mixture, so as to have it well coloured 
 on its under as well as its upper surface ; because, if the particle 
 is placed on the slide before the solution, the part adhering to the 
 glass cannot be coloured. The particle of the sputum should not 
 be larger than i/4th to i/srd of a grain of millet; otherwise, 
 when pressed between the two slips, it will press out from under 
 the cover-glass, and consequently the best specimens, which are on 
 the edges, will be lost, or the preparation will prove too thick for 
 investigating the most delicate parts. 
 
 The particle of sputum, taken with a bent needle, should be 
 immediately put into the colouring mixture, pressed down for a 
 short time with the needle, then left for two hours under a watch- 
 glass ; a drop of distilled water may be added to it, if a weaker 
 colouring be desired ; or, in hot weather, to prevent hardening or 
 evaporation. 
 
 After the required time has passed, take the sputum with a 
 perfectly clean needle, put it . in a watch-glass containing distilled 
 water, and, by gentle agitation, it will be freed from the granular 
 deposits. Meanwhile the slide must be well cleansed. Then put 
 upon it a drop of distilled water, in which the particle is to be 
 again immersed. By using glycerine we cannot obtain the proper 
 thinness of the preparation, and many features would appear to be 
 altered and colourless. Glycerine can ultimately be substituted 
 by capillarity to preserve the preparation, the upper surface of the 
 cover-glass, made greasy by the immersion, being washed first with 
 benzine and then with water. 
 
 In the preceding Memoir, we recommended the spreading and 
 thinning of the particle of the sputum with needles ; but for our 
 present investigation it would not be advisable to do so. It was, 
 perhaps, owing to this that we then lost some details, especially ^ 
 with regard to the fructifications of Leptothrix. In fact, spreading 
 with needles must destroy the relations of continuity and contiguity 
 necessary to preserve intact the said fructifications. 
 
AND CONTENTS OF THE MOUTH. 93 
 
 To thin the preparation as much as possible, we must place 
 upon it the cover-glass, well centred, and let it act by its own 
 pressure. In this way the mucus becomes thinned, and spreads 
 slowly without altering materially the elements involved in it. 
 When the preparation has become partially thinned, press down 
 lightly and gently with the rod and leave it undisturbed, and so 
 on until the very slender film of the coloured matter has in every 
 direction neared the edges of the cover-glass ; care being taken in 
 these handlings to strain off the redundant liquid without wetting 
 the upper surface of the cover-glass. 
 
 The preparations so mounted, in water, are in all parts very 
 transparent, clear, and bright ; they do not exhibit superposition 
 of elements or corpuscles, and even in hot weather, keep well under 
 the microscope for several days, if care be taken to wet now and 
 then one of the edges, as we suggested in the first Memoir. Even 
 the bacteria in the centre of the particle of sputum which, were 
 not at first reached by the colouring medium become in time gra- 
 dually coloured, and these weak colourings are often very useful 
 for examining minute details. 
 
 In order to obtain fructifications of Leptothrix from sputa, it is 
 necessary to take the particle of sputum, not from the bottom, but 
 from that side of the tube where, perhaps accidentally, a vertical 
 streak of the mucus, slender and adhesive, has been left, as we 
 shall see later. 
 
 For researches on fructifications, powerful immersion objectives 
 are required, so as to be able to detect all details. We have used 
 a No. II objective (i/i8th homogeneous immersion) of Hartnack. 
 
 Collection and Preparation of the Contents of the Month. 
 
 We shall especially deal with \hz patina dentaria (surface of the 
 enamel, tartar), saliva and patina of the tongue. 
 
 Patina Dentaria.— The deep layers of this patina contain almost 
 exclusively bud-growths and knots of large filaments which take a 
 brilliant colour with the solution of iodine. The fructifications, 
 on the contrary, are simply found on the superficial layers, so that 
 for their investigation we must possibly remove the upper surface 
 of the patina. By scraping the tooth with a small instrument or the 
 
94 BACTERIA IN THE SPUTA 
 
 nail, too much tartar is removed, which must be disintegrated 
 twice over j first to Select a particle, and then to thin and spread 
 it on the slide, I have best succeeded with a bent needle, first 
 scaling lightly the labial surface of the tooth, and carrying the very 
 minute particle of the patina on to the slide (on which was placed 
 a drop of aniline water), and then scraping likewise the interstice 
 between the teeth, to place on the slide another similar particle. 
 In order not to spoil the fructifications, the scraping should be 
 made neatly from top to bottom. This manipulation should be 
 done in the morning, fasting, and cannot give good results, in 
 those subjects who habitually clean their teeth with tooth-brush 
 and powders. 
 
 Thus we get within the drop of aniline water two tiny islands 
 of Leptothrix, but they are yet too thick to be reduced to the thin- 
 ness required. We shall break them with the needles, so as to 
 carry on to the slide only a kind of whitish dust, which will occupy 
 a third of the area to be covered. Having done this, we shall add 
 with the rod another drop of aniline water, and soon after, with 
 a different rod, a drop of gentian violet solution. We shall speak 
 of the other colourings in Section 4, the treatment being nearly 
 th6 same. The preparation is kept for a little while under a glass, 
 and when it appears to the naked eye to be sufficiently coloured, it 
 is mounted without washing, which, in this case, would take away 
 all the fragments, the fructifications set free, and the bacilli floating 
 about each tiny island. 
 
 When we wish to examine together the saliva and t&e patina 
 dentaria, instead of placing a drop of aniline water previously on 
 the slide, we will put on it a drop of saliva in the manner we shall 
 indicate by-and-by ; the rest of the treatment is the same. How- 
 ever, the preparations without the saliva turn out clearer and 
 thinner ; whilst those mixed with it cannot become thin enough, 
 owing to the resistance presented by cumuli of large buccal epi- 
 thelia, which place themselves like supports between the slides ; 
 still worse when one of those cumuli happens to be on a bed of 
 Leptothrix. Naturally, in the preparations mixed with saliva, the 
 rich cumuli of Diplococci are more abundant, whilst they are 
 seldom found on those of the patina only. 
 
 Lastly, the cover-glass is put on and gently pressed down with 
 
AND CONTENTS OF THE MOUTH. 95 
 
 the rod, until the layer of the Leptothrix, spreading in every direc- 
 tion, reaches the edges of the cover, and the redundant liquid 
 trickles down by its side. The preparations, stained with aniline 
 colours, become a little hard (perhaps owing to the alcohol in it) ; 
 but yet they get thinned enough. Those treated with the iodine, 
 and especially those treated with picric acid, or the colourless ones, 
 being more flexible, become more easily thinned. 
 
 In order to see the fructifications in a fair quantity and dis- 
 tinctly, it is necessary to wait some time. Naturally, those on the 
 edges of each tiny island are seen more distinctly, but all do not 
 show them, because rhany of the islands contain only filaments or 
 bud-growths. Some are wasted, and others overlie one another, 
 mixing their relative edges together. Now, in those edges with 
 exuberant fructifications, these at first are close together ; but the 
 preparation in time partially dries up, and on pouring on it a fresh 
 drop of distilled water the cover-glass slightly rises, and then the 
 tufts of ears begin to open ; the ears separate one from another, 
 and (in preparations with picric acid) several may be counted in 
 the visual field. Such observations can be continued for several 
 days by adding to the preparation from time to time a drop of 
 distilled water. 
 
 To demonstrate the productions by points, the method of 
 working must be modified, as we shall see in Section 4. 
 
 Saliva. — To get the saliva as far as possible free from froth 
 and air-bubbles, the following plan should be adopted : — Let the 
 patient before tasting food spit on a well cleaned glass rod, held 
 horizontally; then incline it So that the saliva shall run down 
 towards the hand, parting, in its course, from the small air-bubbles, 
 which, being lighter, adhere to the rod and stop the froth. When 
 a drop of saliva has gathered on the handle, it may be taken off 
 by the point of a second rod and carried directly on to the slide 
 (if we wish to examine it with the patina dentaria and then mix 
 them together); or, if we wish to examine them separately, it 
 should be immersed in a drop of colouring mixture, already placed 
 on the slide. As soon as the preparation is sufficiently coloured, 
 it should be covered without washing. If the colouring is very 
 weakj'With slight vestiges of colour, the vibrating motion in the 
 salivary corpuscles will be better detected. 
 
96 BACTERIA IN THE SPUTA 
 
 The Cuticle of the Tongue— On an empty stomach, the surface 
 of the tongue is scraped with a spatula or tongue-scraper. We 
 take a small particle of the product and put it in the aniline water 
 previously deposited on the slide. Afterwards it is carefully divided 
 and coloured like the patina dentaria. However, owing to the 
 superposed epithelia, these preparations can never be properly 
 thinned, and therefore are only in a few points rendered clear 
 enough for investigation under high powers. 
 
 We need not deal separately with the nasal mucus and the 
 smegma-balano-praputialis, as the first is treated like the sputa, and 
 the second l,ike the patina dentaria. We shall only notice that the 
 preparations of the smegma do not become satisfactorily coloured 
 with aniline dye, on account of the abundant precipitation, due to 
 fats ; it is better to make use of iodine or picric acid. 
 
 § HI- 
 OPINIONS HITHEBTO HELD RESPECTING THE MICRO- 
 ORGANISMS OF THE MOUTH, according to Miller. 
 
 History and General Facts. 
 
 WHEN I for the first time began to examine the fructification 
 of Leptothrix, of which no mention had hitherto been 
 made in the ordinary text-books of microscopy, I, of course, 
 consulted several of the special publications quoted in the Biblio- 
 graphical Appendix, in order to ascertain whether others had des- 
 cribed these fructifications before; but my search was fruitless. 
 It is true that Robin believed in their existence, as appears 
 from the quotation at the beginning of this Memoir; but the 
 granules mentioned by him are simply the reserve gemmules of 
 this microphite. However, I should not have been induced to 
 treat on this subject so early, had I not unexpectedly met with a 
 very conspicuous specimen of such fructification in some pulmo- 
 nitic sputum (see Fig. i6). I then wished to acquaint myself 
 with the last work of Miller, already quoted, and on perusing it, 
 I found that he had not even touched on that point ; but, on the 
 
AND CONTENTS OF THE MOUTH. 97 
 
 contrary, his views were, in a sense, diametrically opposed to 
 my own. 
 
 Of the specimen found in the pulmonitic sputum I shall 
 speak in Section 4. According to Miller, the opinions hitherto 
 acquired on the microbes of the mouth would be as follows : — 
 
 In the 3rd chapter of his volume, " The Micro-Organisms of 
 the Buccal Cavity : Local and General Complaints produced by 
 them " (the most complete and most recent work on the subject), 
 the illustrious Berlin Professor goes on to relate the ancient and 
 recent scientific opinions on the microbes of the mouth, beginning 
 with their discoverer, Leeuwenhoek, who, in 1683, discovered 
 first on his own teeth, always kept clean, and afterwards on those 
 of an old man, ^'magna cum admiratione" animalcules, drawn by 
 him (Fig. 10 of Miller), some of which resemble the Comma 
 bacilli,: — "Multa exigua admodum animakula jucundissimo modo se 
 moventia." 
 
 Lebeaume compared the tartar on the teeth to coral formations. 
 Mandl held that the tartar proceeded from the chalky remains of 
 the vibriones described by him, which he thought would be killed 
 by heat, by hydrochloric acid, and alcohol. Bijhlmann was the first 
 to observe and describe the fi.laments of Leptothrix, without, 
 however, giving any opinion as regards their vegetable or animal 
 nature. 
 
 Henle was the first to declare that these microbes were of a 
 vegetable nature. 
 
 Erdl treated the decayed teeth with hydrochloric acid, and from 
 the crown he obtained a kind of delicate, enveloping membrane, 
 composed of parasites. 
 
 Ficinus dealt diffusely with microbes of the buccal cavity, 
 which he held to be of an animal nature, and gave them the 
 generic name of dental animalcules. He described the filaments 
 of Leeuwenhoek and of Biihlmann, the -granules, the epithelia, the 
 corpuscles of mucus, forming the patina dentaria, as well as 
 certain infusoria that accidentally dwelt there. He not only held 
 • the bacteria and bacilli to be real animalcules in brisk motion, but 
 he imagined also that they possessed a mouth. He grouped them 
 with infusoria without cilia, probably shelled, after the types of the 
 genera Paramecium and Colpoda. 
 
98 BACTERIA IN THE SPUTA 
 
 In 1847 Robin thought that the principal micro-organism of 
 the mouth was an alga which he called Leptothrix buccalis. 
 
 Klencke, continuing the researches of Biihlmann and Ficinus, 
 drew the 'dental animalcules, the filaments of Biihlmann (repro- 
 duced in Fig. II of M.); and, following the opinions of Ficinus, 
 he even drew the mouths on their ventral side, as can be seen in 
 the quoted figure. 
 
 Meanwhile, the ideas of Robin were accepted, and Leptothrix 
 was generally held to be an alga or fungus sui generis, excepting 
 by Frey, who, together with Hallier, thought it to be a form of 
 Fenicillium glaucum, and by Tilbury-Fox, who restricted it indis- 
 criminately to a form of Oidium.* 
 
 After these historical hints, Miller goes on to describe his 
 methods of investigations, from which it appears that the author 
 and the preceding investigators have not acted with that neces- 
 sary care and discernment, which has been demonstrated, in 
 collecting and preparing the contents of the mouth ; nor have they 
 instituted comparisons with the sputa. They directed their best 
 attention to the culture of the microbes of the mouth, and, in the 
 study of the patina dentaria, they did not avail themselves' of 
 the immersion methods except in the special examination of 
 certain fragments or isolated microbes and of their cultures. It is 
 no wonder, therefore, that they should not have met with the fruc- 
 tifications and the productions of Leptothrix by points. 
 
 Miller speaks of cultures on agar or on calf's blood serum, and 
 from thousands of specimens he gives the preference to agar pep- 
 .tonised with broth, with an additional 0-5 — i per 100 of sugar. 
 
 The cultures were made from saliva, the tartar of decayed 
 teeth, or scrapings from the surface of the tongue or from dental 
 ulcerations and altered pulps of teeth. The cultures from saliva 
 in the gelatine were negative. Often the bacteria reared in agar 
 did not succeed in gelatine, owing to the low temperature main- 
 tained. 
 
 For a general study, he considers an amplification of 20 to 
 300 diameters sufficient, but for the special morphology he used 
 a homogeneous immersion lens. He speaks of fungi derived from 
 
 * Frey, Bibliography. Tilbury-Fox, by Beale, Work, p. 491. 
 
AND CONTENTS OF THE MOUTH. 99 
 
 external sources, as the Bacilli of tare, potatoes, lactic acid, green 
 pus ; and also of the Micrococcus tetragenus, of Mycoderma aceti, of 
 Staphylococcus pyogenes, aureus, and albus, to the effect of excluding 
 them from the cultivations. But he touches again upon some of 
 these forms in Chapter IX., as we shall see later on. 
 
 Miller classifies, afterwards, the microbes really constant, or 
 primary, of the buccal cavity, into six different species. Of these 
 we give a list, putting our denominations to each species of Miller. 
 
 Synopsis. 
 
 Miller : Vicentini : 
 
 I — Leptothrix buccalis (L. Leptothrix buccalis. 
 innominata). 
 
 2 — Bacillus buccalis maximus. Fragments of stumps. 
 
 3 — Leptothrix buccalis maxima. Stumps. 
 
 \—Jodococcus vaginatus. Special sheaths of bacteria pro- 
 
 ceeding from some reserve 
 gemmules. 
 
 S* — Spirillum sputigenum. Pointed or virgulated Bacilli 
 
 (copulatory organs ?) 
 
 6 — Spirochcete dentium. Spirillum (fragments of very slen- 
 
 der filaments). 
 
 The above species or types are found in every mouth. The 
 Spirillum sputigenum is sometimes found, in almost a pure culture, 
 in the carious cavities ; and the Spirochcete dentium near the gums. 
 The cultures from substances mixed with saliva, acids, alkalies, 
 product of caries, dental mucus gathered out of the ebullition 
 of .decayed teeth, do not succeed ; some simple cultures, however, 
 give productions of 15 or 20 articulations of certain species, but 
 these cannot be reproduced a second time. 
 
 I will now sum up the characteristics which Miller attributes to 
 these six primary species, beginning with Leptothrix innominata. 
 
 Leptothrix buccalis (Leptothrix innominata). 
 
 The name of Leptothrix buccalis is attributed to Robin, who 
 applied it in general to all buccal microbes. Hallier, Zopf, and 
 all others who wrote on this subject conformed to his opinion, 
 maintaining that the motile bacteria are spores of Leptothrix in 
 
100 BACTERIA IN THE SPUTA 
 
 activity, and cocci and bacteria not motile are wandering spores 
 in a quiescent state. 
 
 Leber and Rottenstein found the fine violet staining with iodine 
 and the acids to be a characteristic of buccal microbes (included 
 by them in a single species of Leptothrix). Miller, however, 
 considers such forms to be different species, although they show 
 the same behaviour towards staining re-agents, for the reason that 
 the filaments of Leptothrix are not articulated (as stated by Robin). 
 On the contrary, the bacilli or filaments under iodine re-agents are 
 articulated. Now, can this distinctive character be sufiScient to 
 classify a species ? I shall show later that, besides articulations, 
 bacilli may be found, or knotty little, rods in continuity with small 
 chains, and the latter with fertile filaments (not articulated), as in 
 Figs. 9 and lo. This demonstrates the fact that such filaments or 
 bacilli, either articulated or not, or simply in beaded forms or 
 chains, do not constitute a difference of species. Moreover, the 
 filaments more woody and articulated, found in the lower layer of 
 the patina, or, owing to the friction, are left exposed, so that the 
 remaining stems become thick and hard, and their internal 
 gemmules give rise to enclosed bacteria. Vignal was not of a 
 different opinion in holding a fungus of the mouth, which he had 
 reared, to be Leptothrix, although it showed transverse segmenta- 
 tions easily discernible in aniline. Likewise, Miller, in his article 
 on Leptothrix gigantea, as we shall see later, entertained the same 
 opinion. 
 
 Quite recently, the dumb-bell bacteria and other fungi of the 
 mouth were identified with Bacterium termo by Stockwell, Clark, 
 and others. In the preceding Memoir we held this identity as 
 plausible ; but Miller positively rejects it because the title of Bac- 
 terium termo would embrace a mass of very different forms and 
 species. He likewise rejects the title of Leptothrix buccalis (given 
 by Robin) and of dental filaments (by Biihlmann), names which 
 Miller would have banished from microphitology. However, he 
 would give the name of Leptothrix innominata to those little-known 
 filaments which appear to constitute a group or a species differing 
 from other fungi. It is clear that the author alludes to our fertile 
 filaments, making of them a separate species. 
 
 The Leptothrix innominata of Miller (the real Leptothrix akin 
 
AND CONTENTS OF THE MOUTH. 101 
 
 to fructifications to our knowledge) is to be found in the patina 
 dentaria {materia alba of Leeuwenhoek) constantly, but in different 
 quantities in every mouth. It is, of course, very scarce in well- 
 cleaned teeth, because, lying on the top of the tufts, it is the 
 first to be removed by friction. The patina dentaria exhibits large 
 and small heaps of round granules, and, on the edges, slender 
 filaftients variously bent. The granules were considered as the 
 matrix of the fungus, having been taken for spores of Leptothrix. 
 The author thinks them to be either micrococci foreign to Lepto- 
 thrix, or simply linking points of its filaments. 
 
 The filaments are of varied length, from o'5 to o'8 micro- 
 millimetres, sometimes twisted, usually still, and without articula- 
 tions (but this is not always exact). They generally have irregular 
 contours and appear to be badly nourished or are dead, but we 
 shall see that they contain gemmules and bear spores (ears ?). 
 In their surroundings there are also numerous shorter filaments 
 or small rods, which the author thinks might be simply fragments 
 of longer filaments, or cellules of the fungus not having yet 
 reached their full development. 
 
 By using the solution of iodine, slightly acidulated with lactic 
 acid, under a power of 350 diameters, epithelia and clusters of 
 micrococci can be detected intermixed with them (Fig. 12 of 
 Miller), and even various forms of little rods and filaments of 
 Leptothrix lightly tinged with yellow. Other larger bacilli become 
 tinged with deep violet, and these are called by the author Jodo- 
 coccus vaginatus and Bacillus buccalis maximus. 
 
 The same author again speaks of the Leptothrix innominata on 
 page 203. He declares positively that it cannot be cultivated in 
 every medium. This is true, if he intends to speak of the ordinary 
 culture media, but not equally so with regard to sputa, in which 
 are to be found not only natural cultures, but even the most vigor- 
 ous fructifications of this parasite, as we shall see later on. 
 
 Other Primary Micro-Organisms of the Mouth. 
 
 Bacillus buccalis maximus.— This consists of isolated bacilh or 
 
 filaments, more frequently of bundles. The filaments of the 
 
 length of 30—50 micro-millimetres (Fig. 13 of Miller, stained 
 
 with the acidulated solution of iodine) are distinctly articulated. 
 
102 BACTERIA IN THE SPUTA 
 
 The isolated bacilli or articulations are from 2 to lo micro-milli 
 metres; their thickness is from i to 1-3 mm. Of the primary 
 fungi of the mouth this is the greatest ; all its parts cannot be 
 coloured with iodine (Fig. 14 of Miller to be compared with our 
 Fig. 2, m, m, n, n, n, q, r, and with Fig. 5, a, b, c, d). It is not 
 detected in the dental tubuli, probably because it is too big to 
 penetrate there. 
 
 Miller holds it to be of a different species from Leptothrix 
 innominaia, from its size, and through possessing segmentations or 
 knots, and not bending in zig-zag ; and also on account of its 
 very strong behaviour towards iodine re-agents. But these points, 
 very feeble in themselves, are not even constant. Take, for 
 example, the specimen drawn in Fig. 9, d, in which the filament 
 of Leptothrix (fertile filament not segmented) springs from a por- 
 tion of the chain which rose on the top of a large, knotty filament. 
 If these large filaments appear in stumps or in separate networks 
 it is owing to the growth of superior vegetation and the subse- 
 quent rubbing off of the latter by friction. 
 
 I shall presently show that Leptothrix living in the water is abso- 
 lutely identical with this Bacillus maximus of Miller. 
 
 I have, however, to observe that very distinct segYnentations 
 exist sometimes, even in the most slender filaments. In Fig. 5, g, 
 I observed an opaque segment, deeply coloured in the middle, 
 with two clear, pale segments at the ends. This filament, com- 
 pared with bacillus d (same figure), hardly gives the thirtieth part 
 of its thickness. Miller himself, in his previous work of 1883 on 
 Leptothrix gigantea, describes small rods and cocci within the 
 sheaths of filaments of the said Leptothrix (Figs. 2, 3, and 4 of 
 relative plate). In Fig. 5 he even drew a cumulus of small rods, 
 sprung out of the filaments through bursting. The very slender 
 filaments drawn in Figs. 2, 3, and 4 are also articulated. 
 
 Leptothrix buccalis maxima.— The same author is not certain 
 whether this is a really separate species or a younger form of the^ 
 same Bacillus buccalis maximus. The only difference consists in 
 its resisting an iodine reaction, and in the greater distance of its 
 articulations. 
 
AND CONTENTS OP THE MOUTH. 103 
 
 Jodooocous vaginatus (Fig. 15 of Miller, reproduced in our 
 Fig. 7) is found rather plentifully in uncleaned mouths. Miller 
 found it only in two children. It is formed of rom four to ten 
 cellules or nuclei, seldom more, placed obliquely in chain, and 
 having the shape of little flat or round shields, or tetrahedrons. 
 The chain is either bare, as in /, /, of Fig. 2, or sheathed. The 
 sheath, large, 075 mm. (I have found some even larger) is colour- 
 less or takes a yellowish colour, with a long iodine saturation. The 
 nuclei are tinged with dark violet. At times the sheath is broken, 
 the place of the nucleus is empty, or it has dropped off. 
 
 I shall not repeat what 1 demonstrated in Section I. with 
 regard to these chains — of their envelope tardily discovered, of 
 the presence on the same of curved diplococci, morphologically 
 identical with the gonococcus of Neisser. I shall only state that, 
 as such forms are found, even of greater dimensions, in urine, and 
 being intermixed with filaments and other forms of Leptothrix, 
 their relation with this parasite, which is most widely spread, 
 becomes still more probable. It is, perhaps, the only one which 
 lodges simultaneously in the mouth and in the external genito- 
 urinary mucous membrane. 
 
 But, even admitting the Jodococcus vaginatus of Miller to be 
 entirely different from the small sheathed chains described by us 
 in Fig. 4, their entity as a distinct species still remains doubtful. 
 The same author, in the quoted work of 1883, described and drew 
 (annexed plate, Fig. 8) some articulations divided in two and in 
 four small cocci, arranged, as in the Jodococcus vaginatus, by 
 couples or tetrahedrons. He even found in the pig whole filaments 
 of Leptothrix, made up with a series of those cocci, as we shall 
 see later. 
 
 Spirillum sputigenum (Figs. 16 and 17 of Miller and our Fig, 
 g^ ^). — It is found in every mouth, and mostly upon unclean 
 teeth. We should note this circumstance, as it corroborates our 
 views. Cleaning the teeth destroys, every time, the pseudo-inflo- 
 rescences or productions by points, from which would rise those 
 Comma bacilli, together with the spindle-like and snake-like bacilli. 
 We shall see, in fact, that the simple act of mastication is apt to 
 destroy those pseudo-inflorescences, so that we must look for them 
 
104 BACTERIA IN THE SPUTA 
 
 on empty stomachs. Is it to be wondered, then, if their products, 
 and chiefly the Spirillum sputigenum, are so scanty in those persons 
 who regularly clean their teeth ? Miller goes on to say that in 
 unclean mouths they are numberless. 
 
 These microbes are in the shape of small Comma rods, endowed 
 with very quick, screw-like movements. When linked by twos, 
 they form themselves into small snakes, like the letter s. 
 
 Lewis, as we have already said, identified this microbe with the 
 bacillus styled Cholerigenus. However, its presence in the mouth 
 had been detected before, and by a few attributed to fragments of 
 Spirochete. Clark held it to be the cause of caries, for it pene- 
 trates the dental tubuli as shown in Fig. 17 of Miller. This author 
 excludes every relationship between the Spirillum sputigenum and 
 the bacillus Cholerigenus, judging from the culture which for the 
 latter is positive whilst for the Virgula of the mouth it is always 
 negative. However, this is a new confirmation of our views, for, 
 if the Commas in question are real organs or copulative filaments, 
 it is quite natural that, whilst they fecundate the articulations or 
 the spores, they should be incapable of reproduction themselves.* 
 
 In addition to the Spirillum sputigenum. Miller classifies two 
 other types of curved filaments : one short, massive (Fig. 32), 
 and motile, which liquifies gelatine \ the other slender, still, and 
 more curved (Fig. 18). In growing old, it gives rise to a small 
 
 * After having completed our work, we found in the Lancet of June, 1890, 
 an important article by Dowdeswell, on the Comma bacillus of cholera. He 
 touches on the widely-spread hypothesis that those Commas may be only frag- 
 ments of spirilla, and then he describes three cycles in their evolution : — 
 
 A, Commas or fragments of spirilla which may or may not end in sporules ; 
 
 B, Active cellules, with cilia and amoeboid forms ; then round quiescent cel- 
 lules (sporanges) ending in minute sporules ; C, Active filaments ending in 
 sporules of the first generation. The author, however, has not succeeded in 
 reproducing, by any method, the normal Commas, vital and reproductive for 
 themselves. He affirms that cholera bacilli are to be found, normally, in the 
 large intestine of the guinea pig, and that the subcutaneous inoculation of a large 
 number of such bacilli really produces choleriform symptoms in that animal ; 
 but it is not a true cholera infection. Considering the high temperature that 
 accompanies it, it is a true septicaemia. The fact, then, that the author nas 
 found forms of Leptothrix in the Comma bacillus cultures, goes still further to 
 support our views. Dowdeswell, Note on the " Morphology of the Cholera 
 Comma Bacillus," in the Lancet, 1890, Vol. i., page 14 19 — 23. 
 
AND CONTENTS OF THE MOUTH. 105 
 
 chain of cocci. The author has not detected any spores. Although 
 this microbe grows in the gelatine, it does so in a different manner 
 from the cholera bacillus. 
 
 SpiroohsBte dentium (Fig. 20 of Miller and our Fig. 6, c) is not 
 found in the decayed teeth, but on the edge of the gums, together 
 with the Spirillum sputigenum (affection of the gums). It exhibits 
 'spires. 8'25 mra. long, of various thicknesses. The more slender 
 ones hardly become coloured. The author himself doubts whether 
 the slender spires constitute a separate species, because he con- 
 siders the largest amongst them to be akin to the Spirillum sputi- 
 genum. Their development is probably unknown. " We know 
 little or nothing (he says) about the vital conditions and their 
 manifestations, such as fermentation, pathogenic action, etc." 
 
 I shall observe that, amongst the materials which came under 
 my examination, 1 have constantly found these spirilla or spiro- 
 chata with the Leptothrix. In the former Memoir on relapsing 
 fever, I spoke of Spirilla and Spirochseta, which, together with 
 filaments, small rods, and other portions of Leptothrix, are pro- 
 duced in infusions of potatoes. In a later paper upon a Dip- 
 lococcus analogous to the Gonococcus of Neisser, in a case of 
 Carcinoma of the bladder, I mentioned the striking Spirochceta 
 that are found in the sediment of bottles filled with the stale water. 
 Together with Zopf, I have retained the name Spirochceta for 
 those spirilla which bend in the middle, and thus mark a 
 sort of transition between the so-called Spirillum and the 
 Spirulina. I may add that, in the sediment of the bottles, 
 the Spirochceta surround the tiny islands or lumps of a stringy 
 microphite, which, in its natural state, resembles the filaments of 
 the Leptothrix gigantea from the dog, especially the more slender 
 ones, drawn in Figures 2, 3, 4 in the already mentioned article of 
 Miller. On the other hand, the intertwined filaments of that sedi- 
 ment, being coloured with aniline, resemble entirely the lumps of 
 the Bacillus buccalis maximus of Miller, after Fig. 13 in the last 
 work of this author. According to Zopf, Leptothrix is a micro- 
 phite that lives in the water.* 
 
 * Zopf, Die Spaltpihe, Breslau, 1883, page 80.— Vicentini, On a diplo- 
 coccus analogous to the gonococcus of Neisser, found in urine, in a case 
 of Carcinoma of the bladder— Vol. XLin., DegK Atti Delia R. Accademia 
 di 'Napoli, 1889. 
 
106 BACTERIA IN THE SPUTA 
 
 I have already stated that, in the midst of those masses 
 of filaments of the sediment in the bottles, there are preserves of 
 Vorticellas. 
 
 Now, it is not improbable that the Spirochmta rise there from 
 the more slender filaments of the Leptothrix gigantea, and in 
 the mouth from those of Leptothrix buccalis, as I shall show 
 later ; and that the Leptothrix gigantea and the Bacillus buccalis 
 maximus, or Leptothrix buccalis maxima of Miller, may be the 
 same thing. I have remarked that, in the sediment from the 
 bottles, the vorticellas and filaments are never wanting ; whilst the 
 Spirochceta are found there only in summer, and then in brisk 
 motion. Hence, we may infer that the temperature may greatly 
 influence the production of the Spirilla and the Spirochaeta, as 
 Miller had argued from the cultures, and as their abundance in 
 the mouth would prove. 
 
 I shall refer later to the Spirilla that, in certain cases, I 
 have seen accompanying the Leptothrix of the prepuce and the 
 urethra. 
 
 Secondary Micro-Organisms. 
 
 Leptothrix gigantea (Fig. 21 of Miller). — This form was found 
 on the teeth of a dog affected with alveolar pyorrhoea. It 
 was also found on other carnivorous and herbivorous animals. 
 It exhibits a vigorous development ; has a bushy shape, like the 
 Crenothrix, the stems of which deflect at the top, with cocci, small 
 rods and filaments sometimes alternated on the same line. The 
 stems are polymorphic of varying thickness. Some increase in 
 size towards the top ; some are partially or wholly spiral ; and 
 others double and twist as in Spirulina (Fig. 12 of the plate 
 annexed to the article of 1883). 
 
 In this article, published in the Berichfe der Deutschen 
 Botanischen Gesellschaft, are to be found more particulars upon 
 the microphite in question. The oldest filaments are articu- 
 lated ; some are spiral and mutually twisted at the base (Fig. i, 
 E, a). The articulations of the more slender filaments cannot 
 be detected without colourisation. There are sheathed filaments 
 whose internal articulations sometimes evacuate and form cumuli 
 of little rods of different sizes (Fig. 5) ; others become compressed 
 
AND CONTENTS OF THE MOUTH. 107 
 
 or swollen, or deviate from the axis (Fig. \, A, B, C). By staining 
 process we find, particularly in the pig, articulations subdivided 
 into two or four cocci ; or groups of two or four, which at times 
 occupy the whole length of the filament (Fig. 8). In the tufts 
 can be seen straight and tortuous filaments. The bending is 
 graceful and extended. These are styled by the author, Vibrionicox 
 Spirochatic forms (reproduced in our Fig. 8). In the more slender 
 filaments we can observe the transition from the vibrionic to the 
 spirillic form upon a single filament (Figs. lo and ii). In the 
 spirillic forms of these slender filaments, by means of a proper 
 colouration, the articulations can be detected. 
 
 In 1883, Miller suspected that the same slender spires of the 
 Patina Dentaria would rise from the fragments of long and slen- 
 der tortuous filaments (Figs. 13 and 17). In the dental Spirochgeta 
 there is no trace of articulations, but in the marshy Spirochseta, 
 through the weak colouring of Zopf, are detected several articula- 
 tions in every filament (Fig. 20). Hence, then, the author inferred 
 that relations existed between the Spirochseta and other microbes 
 of the mouth. But now, in his work of 1889, he seems to have 
 discarded his former theory of unity of forms. 
 
 Miller is unable to say whether the Leptothrix, detected by him 
 and by others in. the tame herbivora and in the pig, is identical 
 with that which lodges in the mouth of man and of the carnivora. 
 He says that it would be necessary to institute pure cultures. 
 Anyhow, it is important to admit the existence of Leptothrix, even 
 in the herbivora. Whilst before it was held that Leptothrix lodged 
 exclusively in the mouth of carnivorg, Zopf had already admitted 
 its existence in the herbivora.* It is true that it is less frequent 
 in the herbivora ; but, if we are not mistaken, it must be taken 
 into consideration that these, and especially the ruminants, are 
 continually using their teeth, so that the parasite cannot thrive 
 there at ease.f 
 
 * Comil and Babes' quoted work, page 135 ; Zopf already quoted. 
 This author thinks it probable that Leptothrix buccalis originated from the 
 external world, especially through water and food. We have already mentioned 
 the Leptothrix found by us in the sediment of a bottle of water ; but how can 
 we explain the Leptothrix in genito-urinary passages ? 
 
 t Miller, Uebereinen Zahn Spaltpih, Leptothrix gigantea (Berichte der 
 ' Deutschen Botanischen Geselhchaft, Heft S, 1883, p. 221). 
 
108 BACTERIA IN THE SPUTA 
 
 Fungi of the Mouth which may be stained blue and violet 
 with Iodine Colours.— Besides the Bacillus buccdlis maximus and 
 the Jodococcus vaginatus. Miller points out three others with a 
 striking iodine reaction, viz. : — {a) Jodococcus magnus (Fig. 22), 
 which colours violet ; (V) Jodococcus parvus also stains violet ; and 
 {c) another Jodococcus, which takes the rose colour with iodine, 
 and can be cultivated a first time, but it does not reproduce in a 
 second culture. 
 
 Fungi of the Mouth which can be cultivated.— Some of these 
 are not pathogenic ; some of a doubtful pathogenic efficiency. In 
 Fig. 23 of the author are depicted several various types of these 
 bacteria of the mouth ; in a, c, g, screw forms ; in b, cocci ; in 
 d, small rods ; in <?, a sheathed chain of cocci ; in i, a long 
 chain (but these are numberless in the patina of the tongue, so 
 that we believe it to be impossible to separate them from Lep- 
 tothrix) ; in /, chains of small rods ; and in k, thread forms (to 
 which the same remark applies). 
 
 Miller himself considers it a confusion, difficult to clear up 
 without immense labour, as all the bacteria of the external world 
 can form a nidus in the mouth. The author depicts 19 out of the 
 22 species which he separated in 1885, viz., from Fig. 24 to 27, 
 cocci; 28 to 29, small rods ; and 30 to 31, still longer rods ; in 
 Fig. 32, a Vibrio viridans that liquefied the gelatine and produced 
 a green colouring matter ; a spirillum (Fig. 33) ; long threads 
 (Fig. 35) ; and small chains, partially sheathed (Fig. 34). 
 
 We shall omit seven other types obtained from simple cultures. 
 
 Miller afterwards points out the investigations of Vignal, which 
 he finds fit and accurate. Such investigations led to the cultiva- 
 tion of 1 7 species of fungi in pure cultures ; some identical with 
 known fungi; others unknown. The more frequent was Bacterium 
 termo ; then come Bacillus ulna ; then the Bacillus of potatoes ; 
 three other unknown Bacilli ; the Bacillus subptilis ; then the 
 Staphylococcus pyogenes albus; then the aureus; five other unknown 
 Bacilli ; and, finally, the Leptothrix. 
 
 From that view, there would result for Vignal a great prepon- 
 derance of bacilli; for Miller, on the other hand, of cocci. Miller 
 attributes this divergence of results from Vignal having used the 
 
AND CONTENTS OF THE MOUTH. 109 
 
 gelatine, on which not all of the above mentioned microbes thrive, 
 Black, through his own researches, has found in the contents 
 of the mouth, Streptococcus continuosus, Staphylococcus medius. 
 Staphylococcus magnus, Coccus cumulus minor, and Bacillus 
 gelatogenes.* 
 
 We have not been able indeed to convince ourselves that, from 
 those experiments of cultures, a basis may be established for the 
 identification of the species of bacilli or bacteria, owing to their 
 unlimited polymorphism, according to the various nourishing media 
 and the diiferent conditions of their surroundings ; so that, wher- 
 ever the decisive proof of the fructification is wanting, the results 
 of the cultures for creeping or immersed vegetation cannot be 
 freely accepted as the. same. In doing so we might run the risk 
 of multiplying the species and the true or supposed pathogenic 
 properties of bacteria, ad infinitum. 
 
 Different particles or cellules of the same organism may, \n 
 fact, need various nutrient media ; they may liquefy certain sub- 
 stances, and separate others of a very different nature ; as, for 
 instance, the bony corpuscle, the hepatic cellule, the blood cor- 
 puscle, the Renal Epithelium, etc. The poison of insects or 
 snakes is not difi"used throughout their organism, but it is secreted 
 by special glands. Likewise, in the higher upgetation, the leaves, 
 the roots, the buds, the seed, etc., often contain principles or alka- 
 loids totally different from the other parts. And the same particle 
 or cellule can assume various forms according to its stage of deve- 
 lopment, its surroundings, or nourishing pabulum. 
 
 Furthermore, the virulence of bacteria may decrease or disappear 
 in certain cultures ; increase or reappear in others. Now, if the 
 virulence constitutes for itself a truly specific character at every 
 step, we shall have to admit, in those microbes, changes of species; 
 and thus, in order to defend the true and well-thought pleomor- 
 phism, we must necessarily fall into an illimited and unlikely one. 
 
 However, referring to the six species of micro-organisms 
 admitted by Miller in the cavity of the mouth, we, with due 
 respect for the author, cannot abandon the opinion (which, 
 although old, is not ours) of the fundamental unity of all 
 these forms, and their derivation from a single species, the Lep- 
 * Miller, Die Mikroorganismen der Mundhiihle, etc., 1889, pp. 43 — 75. 
 
110 BACTERIA IN THE SPUTA 
 
 tothrix buccalis of Robin ; of which, in the following section, we 
 will give our description in its principal biological phases. At 
 most, some doubt might be entertained ia regard to the Spirilla 
 or Spirochaeta, whether to hold them as a species, or belonging to 
 a distinct species ; but their relationship with Leptothrix was 
 recognised even by Zopf, who, on this point, wrote : " In certain 
 cases, the filaments (of Leptothrix), either on a given part or in all 
 their length, take a spiral shape, and the fragments of those fibres 
 form the Spirilla, Vibriones, and Spirochcetee." * 
 
 Miller himself in mentioning his studies, made in 1883 
 upon those of Zopf, also admitted the derivation of the Spiro- 
 ckcetm of the human mouth from the twisted threads of Lepto- 
 thrix, of which he gave a representation of the fragments in the 
 figures 13 to 20 of his plate; and, in the description of the 
 branching filaments of Leptothrix gigantea of the pig (Figs. 6 to 8) 
 and of the sheep (Fig. 9 to 1 2), he represented by the side of 
 straight filaments other vibrionic and spirillic ones.t 
 
 We have besides, by our observations, pointed out that the 
 Spirilla or Spirochceta are constantly united with Leptothrix, even 
 in the infusion of potatoes and in stale water (in a warm atmo- 
 sphere). Later on, we shall see that they are also found in certain 
 urinary mucous flakes, always accompanied by vigorous fructifica- 
 tions of Leptothrix. 
 
 Pathogenic Fungi of the Mouth. 
 
 We conclude this short extract by summing up the views given 
 by Miller in Chap. IX. of his work upon the experimental infec- 
 tions produced through the inoculation from certain microbes of 
 the mouth, or accidentally lodged in it. 
 
 The-author begins by reminding us that the venomous property 
 of saliva was long before known when, in 1873, Wright and 
 Senator used it for the purpose of inoculation with deadly result. 
 It was suspected that the poisonous effect depended upon a chemi- 
 cal cause. Moriggia and Marchiafava, in 1878, inoculated with 
 the saliva of children dead from rabies. The first who 
 
 * Zopf, loc. cit., page 80. 
 t Miller, Article cit. 1883, pp. 223 — 24. 
 
AND CONTENTS OF THE MOUTH. Ill 
 
 attributed to bacteria the venomous action of the saliva or 
 sputa were Eaynaud and Lannelongue, in 1891. The rabbits 
 that were inoculated with the saliva of a boy who died of 
 rabies, died ; whilst the inoculations with the blood or the buccal 
 mucus were harmless. Pasteur saw two rabbits die after inocula- 
 tion with the same saliva ; and the cultivated microbes exhibited 
 the shape of dumb-bell bacteria in a gelatinous capsule. He then 
 believed he had discovered the cause of hydrophobia ; but Cohn- 
 heim, having noticed the rapidity of the infection, qualified it for 
 septicaemia. 
 
 After these first experiments, Vulpian produced the same infec- 
 tion by inoculating with the normal saliva ; and, in the blood of 
 the inoculated animals, Rochefontaine and Arthaud detected 
 microbes quite identical with the above-named. Sternberg and 
 Claxton confirmed the observations of Vulpian ; but GrifiSn 
 demonstrated that the saliva of the parotides, gathered separately, 
 is harmless ; so that the infection is to be attributed to secondary 
 products of decomposition, formed in the buccal cavity. Gaglio 
 and De Mattei held the same view, and noticed that the saliva 
 became harmless after boiling. 
 
 Fraenkel in 1884 confirmed anew the observations of Vulpian. 
 He found that dogs were more receptive, the mice less so, and 
 still less guinea-pigs. Dogs, pigeons, and fowls were refractory. 
 
 Miller obtained identical inoculations with the excretion of a 
 woman affected with micosis of the tonsils. He separated after- 
 wards from a decayed root five fungi, of which one was distinctly 
 pathogenous. 
 
 Klein and others studied the venomous action of groups of 
 microbes of the human sputa, so that in late years pathogenic 
 properties were gradually attributed to a considerable number of 
 bacteria. 
 
 Pathogenic Fungi not Cultivable.— The author repeats here 
 what has already been said of the primary microbes of the mouth, 
 to demonstrate that Spirillum sfutigenum and Spirochcete dentium 
 are especially not cultivable. Kreibohm found in the patina of 
 the tongue two pathogenic bacteria unfit for cultivation in any 
 artificial medium whatever. Miller, with fungi from a gangrenous 
 pulp of the teeth, obtained a gangrenous product for re-inoculation. 
 
112 BACTEEIA IN THE SPUTA 
 
 Pathogenic Fungi Cultivable.— (a) Micrococcus of the sputum 
 Septiccemia (Figure 96 of Miller). — This is the same bactferium 
 that Pasteur and others obtained from healthy and pathological 
 sputa. Klein was the first to cultivate it. It results in oval cocci, 
 diplococci, and capsulated chains identical with Pneumococcus. 
 Subcutaneous injections with its cultures produce septicaemia and 
 death in 24 — 36 hours. Even the blood of animals inoculated is 
 totally infected; but the infection, once overcome, is contracted 
 no more. Its virulence is attenuated by cultures in milk. This 
 bacterium (continues Miller), lodging constantly in the human 
 mouth, passes from there into the pulmonary tubes ; so that the 
 uncleanliness of the mouth would be the real predisposing 
 condition to pneumonia, in so far as it allows the diplococci in 
 question, deposited there from the air, to multiply. From those 
 views to ours, to the entirely secondary propagation of such bac- 
 teria in the pulmonary products, there is but one step. (See 
 Section 4 of Memoir on Whooping-cough.) 
 
 (p) Bacillus crassus sputigenus (Fig. 97 of Miller, from the 
 blood of a mouse). — This bacterium was found by Kreibohm once 
 on the patina of the tongue and twice in the liquids of the mouth; 
 it is longer than it is broad, but often shorter in its younger articu- 
 lations ; resists Gram's method ; sporifies through heat. Mice, 
 inoculated even with small quantities, die in forty-eight hours, 
 exhibiting myriads of bacteria in their liver and blood. A copious 
 injection of it into the veins kills rabbits and dogs in from 
 three to ten hours from gastro-enteritis. 
 
 (c) Staphylococcus pyogenes albus and aureus, and Streptococcus 
 pyogenus. — These bacteria, held as promoters of suppuration, are 
 brought from outside into the mouth, where they seem to find a 
 fit soil. Black, upon ten specimens of buccal liquids and scrap- - 
 ings from the tongue, found seven times Staphylococcus pyogenes 
 aureus — four times the albus and three the Streptococcus, which he 
 believes with a careful search can always be found. Vignal never 
 came across the true streptococcus ; seldom found the two staphy- 
 lococci. Netter, upon 127 specimens, found only seven times the 
 Staphylococcus pyogenes aureus. Miller also rarely found these 
 micrococci. 
 
 (d) Micrococcus tetragenus (Fig. 98 of Miller).— This micrococ- 
 
AND CONTENTS OF THE MOUTH. 113 
 
 cus, relatively frequent, was first found by Koch and Gaffky in the 
 cavities of a consumptive subject ; it is more frequent in tubercu- 
 losis. Biondi found it in three cases out of five. It is in the 
 shape of cocci disposed two by two or four by four ; with gelatin- 
 ous matter interposed, it does not liquefy the gelatine. White 
 mice and guinea pigs inoculated with its cultures die in from three 
 to ten days ; those inoculated with sputa perish in from four to 
 eight days. Their blood and their organs are infested with micro- 
 cocci. 
 
 Fungi of Biondi.— This investigator has isolated from the 
 mouth five different pathogenic bacteria, including the Micrococcus 
 /tf/ra^g^^^'above-mentioned. The four others are : — (e) a Bacillus 
 salivarius septicus, rather frequent ; (f) a Coccus salivarius septicus, 
 obtained in a single case only from a woman affected with a serious 
 septiceeraia in her confinement ; (g) a Streptococcus septicus, similar 
 to that of erysipelas, phlegmon and puerperal Metritis; (h) a 
 Staphylococcus salivarius pyogenes, found once in the sputum of a 
 scarlatinous quinsy. 
 
 The Fungi of Miller.— After many cultures and inoculations 
 from buccal liquids and dental gangrenous pulp, Miller observed 
 more serious effects from the inoculations of the pulp, even 
 in comparison with inoculations of cultures of the same pulp. 
 The effects were varied in different animals and according as the 
 inoculation was made under the skin or the peritoneum cavity. 
 The pathogenic fungi isolated by the author were : — («) a Micro- 
 coccus Gengivcs pyogenes (Fig. 99 of Miller), obtained from alveolar 
 pyorrhoea, cocci that did not liquefy the gelatine ; {k) a Bacterium 
 Gengivce pyogenes (Fig. 100 of Miller), found with the former, in 
 the shape of bacteria four times longer than broad, apt to liquefy 
 the gelatine ; (I) a Bacillus Dentalis viridans (Fig. loi of Miller), 
 obtained from the decayed roots of teeth, in bacteria of irregular 
 forms ; (m) a Bacillus Fulpce pyogenes {^ig. 102 of Miller), obtained 
 from the gangrenous pulps, in bacilli, frequently irregular, isolated, 
 linked together, or in small chains, even with eight articulations, 
 apt to liquefy the gelatine.* 
 
 We cannot further describe these culture experiments. We 
 
 * Miller,, /oiT. cit., pp. 199—220. 
 
114 BACTERIA IN THE SPUTA 
 
 shall only remark that the pneumococci, or theexciters of salivary 
 septicaemia, being constantly found in the contents of the mouth, 
 are to be held, in our opinion, as one of the forms or phase of 
 the microphite or microphites of the mouth (zoogleic phase of 
 Billet). With regard to all the other bacteria we have just men- 
 tioned, it remains to be observed whether they are accidental — 
 i.e., proceeding from without — or normal, although modified in 
 their properties by variable conditions of pabulum or surroundings ; 
 and whether their numerous varieties are perhaps more apparent 
 than specific. 
 
 As regards the experimental inoculations, we shall simply notice 
 the fact that they introduce, under the skin, in the cavity, or in the 
 circulation, all at once, myriads of bacteria. A single centigramme 
 of culture or of polluted material may contain, on an average, 
 twenty-five billions of bacteria, as we have elsewhere demon- 
 strated ; and twenty-five milliards inoculated in a mouse are as 
 fifty or a hundred trillions (Italian measure) inoculated in a man. 
 Is it wonderful if local or general effects may be produced from 
 the sudden irruption of such a mass of parasitical germs, even 
 though in themselves innocuous ? Nevertheless, that is not the 
 real question j the real question is whether the different kinds of 
 bacteria (probably belonging to the same species) have the power 
 of attacking, sponte and on their own account, the human economy; 
 and whether, even penetrating it from their media, in an infinitesi- 
 mal number (as is the case in the ordinary infectious diseases), can 
 they develop in the human species the same effects following upon 
 the inoculation of their elements on the brutes? 
 
 In other words, one is the receptivity through the natural pas- 
 sages and another through the continuous solutions ; one is the 
 slow, successive, and external action of a few germs, another that 
 of the sudden and internal mass of germs. Finally, one is the 
 receptivity of this or that animal species, and another that of the 
 human species. And all this is irrespective of the other more or 
 less questionable points, and especially of the query : If the mor- 
 bific action of the inoculations depends upon the bacteria, as 
 such, or upon the polluted matter in which they are suspended ; 
 or, it may be, upon their products of secretion and decomposi- 
 tion, as is at the present time generally argued. 
 
AND CONTENTS OF THE MOUTH. 115 
 
 But, leaving for the present those complex questions, I will 
 proceed to speak of the morphology and biology of Leptothrix.*- 
 
 § IV. 
 
 REMARKS ON THE MORPHOLOGY AND BIOLOGY OF 
 THE LEPTOTHRIX BTJCCALIS. 
 
 The Four Phases of Leptothrix and particularly the 
 Fructifications by Ears (SpiCje). 
 
 First Phase. — The first phase of the Leptothrix is that of 
 a Bacterium or Bacillus, already described in the Memoir on 
 Whooping-cough and in the first paragraph of the present one. 
 This phase is common to all the other species of bacteria, but 
 which (at least in Leptothrix^ does not represent its whole cycle of 
 life, but only its primordial stage of immersed vegetation, or a 
 vegetation destined to propagate in liquid or semi-solid substrata 
 (media). 
 
 Second Phase. — Examining the cuticle of the tongue, stained 
 with gentian violet, we find there by preference the forms of the 
 second stage of life of this parasite — namely, that of chains, 
 bundles, and masses of intertwined filaments. We have already 
 said that in these preparations the buccal epithelia predominate ; 
 isolated filaments are found there, as well as many specimens of 
 large dumb-bell bacteria of the type//, and/' (Fig. 2), and con- 
 spicuous masses of diplococci. These preparations show that the 
 large dumb-bell bacteria are derived from the diplococci, the two 
 original cocci linking together. The degrees of transition to be 
 observed there are many. The same chains are often surrounded 
 
 * Two fresh works were lately published on this argument : one by David 
 (Les Microbes de la bouche, preMi ifune lettre-priface de M. L. Pasteur), 
 a simple compilation ; the other by Billet (Contribution a Fitude de la 
 Morphologie et du d'eveloppement des bactiriacies ), the result of original re- 
 searches on the natural history of four species of micro-organisms (Cladot/irix 
 dichotoma. Bacterium Balbiani, Bacterium osieophilum, and Leptothrix para- 
 sitica, a parasite of sea-weeds). We shall notice these works later on ; we may 
 at once say that not even these authors hint in the least to the superior 
 PHASES of Leptothrix buccalis, after our type or other's. 
 
116 BACTERIA IN THE SPUTA 
 
 by similar masses of diplococci ; even the entanglements of the 
 type «, u, are not wanting (Fig. 2). 
 
 The chains belong to types c and d (Fig. 2), but are, generally 
 longer, thick, branched off in bundles of 10 or 15, which take up 
 to three visual fields, and exactly represent the disposition of the 
 bundles of filaments. In the same chain small diplococci may 
 alternate with elliptical bacteria and medium-sized dumb-bells — all 
 perfectly equal, which form the largest number. 
 
 In these we have been unable to find fructifications, so that 
 this second phase may be held as analogous to mycelium, or the 
 vegetation, of fungi, partly immersed, partly creeping. 
 
 Third Phase (Incomplete aerial vegetation). — This is generally 
 met with in the tartar of the teeth. Its predominant elements are 
 the large filaments, often very long, bent, and re-united in bundles, 
 taking a brilliant colour with iodine ; and the stumps. T shall 
 not describe these well-known forms, having already done so in 
 speaking of the primary and secondary species of Miller. 
 
 I would rather point out some features, noticeable in a certain 
 number of these filaments, when I have the opportunity of finding 
 them isolated (especially in the unstained preparations), such 
 features not having been considered by previous authors. 
 
 The first is a division into two or three branches (Fig. 9), 
 which I think must be taken for a kind of radical systern of this 
 microphite, for divisions are never detected towards its apex ; and, 
 at its opposite end, a swelling or head is noticed, which is a 
 proper form of points twined upwards (as in b). In a the filament 
 is broken towards the top and exhibits two long roots ; in b, there 
 are three hood-shaped roots. Both appear to be swollen at their 
 ends, like haustoria. 
 
 What can be the use of these barbs or roots, unless to obtain 
 a firm foundation in the ground ? And to what purpose is this 
 firm foundation, unless to support higher forms of vegetation ? 
 
 The other feature is that of the swelling, which we will call 
 apices or heads, being found at the top of the respective filaments, 
 as in b and c, and these are of varied form, at times containing a 
 kind of nucleus. There are others which might be styled knotty, 
 being found along the filament j of these we give no drawing, as 
 they are similar to the preceding ones. 
 
AND CONTENTS OF THE MOUTH. 117 
 
 With the above-mentioned stumps are intermixed almost all 
 the forms of bacteria, bacilli, spirilla, etc., besides a large number 
 of very varied small chains, after the types c, d, k, i, x, and x 
 (Fig. 2). 
 
 The larger size and the facility to colour of these masses and 
 stumps, with regard to fertile filaments, would depend, I think, 
 upon a very natural cause. The friction, removing the points of 
 the filaments and the fertile, filaments, seems to impart a greater 
 development to the remaining stems in the sense of thickness 
 as, for instance, it happens in the pruning of trees, through the 
 retrocession of the ascending sap. 
 
 In the present case, the retrocession of the germinal matter 
 would be the cause of the consecutive increase of the sheath and 
 of the gemmules contained in it. Thus might be easily explained 
 the appearances, so various, of the stumps and their fragments, 
 compared with the fertile filaments ; appearances which induced 
 Miller to make of residual filaments two distinct species, under 
 the name of 'Leptothrix buccalis maxima and Bacillus buccalis 
 maximus, in opposition to our fertile filaments, which for Miller 
 constitutes a third species, the Leptothrix innominata. 
 
 It remains, then, to be seen whether the longer filaments, sunk 
 down and intertwined, of Bacillus buccalis maximus (Fig. 13 of 
 Miller) may not concur, with the above described chains, to form 
 that kind of mycelium or creeping vegetation, upon which is based 
 the aerial vegetation of this micropbite. 
 
 From the tiny islands of the stumps in question, prepared in 
 the manner already described in Section 2, spring at last the fruc- 
 tifications. The bigger filaments, that we would call woody, gra- 
 dually become thinner and pale, showing in their interior countless 
 granules or parietal gemmules. These are the fertile filaments. 
 They may spring either from the proper filaments, with continuous 
 contour, or from the little chains after type d (Fig. 9), which are 
 seen occasionally on the top of those filaments ; and in this last 
 case, instead of a gradual thinning, may abruptly pass from the 
 chain to the fertile filament, as shown in the figure. 
 
 It is by no means uncommon to find parietal gemmules, as 
 Robin mentions the circumstance, and gives a first drawing in 
 Fig. 2h of his volume of 1853, magnified to 800 diameters.* 
 * Robin, Hist. Nat. des Vtgitaux, etc., 1853- (See Bibliography.) 
 
118 BACTERIA IN THE SPUTA 
 
 Fourth Phase {Completed Aerial Vegetation: Fructification). — ■ 
 In Figs. 10 — 13 are reproduced four specimens of fructifications, 
 taken from t\\t patina dentaria. In Fig. 10 are seen three isolated 
 "ears" (spicce ), stainad with gentian violet; in Fig. 11, a tuft, 
 caught as the bird flies, stained with weak methyl violet ; in Fig. 
 1 2, a part of a more vigorous tuft, seen in profile, stained with 
 weak fuchsine ; and in Fig. 13, two fructifications, set on the same 
 tiny island of materia alba, stained with solution of iodine. The 
 Figures 10, 11, and 13 are drawn magnified to 1,250 diameters, 
 and the Fig. 12 to 850 diameters. 
 
 The fertile filaments are at times partially straight, as in Fig. 
 lo; at times bent (even occasionally tortuous, as in Fig. 13), and 
 are sometimes wanting, because the fructifications have been 
 carried away by mechanical force, although grouping together, as 
 in Fig. 12. In Fig. 10 are seen, in a, a, the gemmules of reserve 
 adhering to the walls ; in b are found the little spores properly 
 lodged ; but in b' we notice only five, the others having dropped. 
 In c the penultimate articulations of the stalk appear older and 
 woody ; the last is granular, like the two articulations on the apex 
 of the younger filament, d. 
 
 The sporules, very small, round, brilliantly coloured, show 
 themselves in three vertical rows in all the specimens without 
 exception ; i.e., they never appear in two rows, whence we may 
 infer that they are, in fact, disposed in six longitudinal lines (or 
 series), for, if they were only in four or five, they should, at times, 
 exhibit only two visible rows. 
 
 In turning the micrometer screw, the cylindrical relief of the 
 " ears " appear manifest, for at a higher focus the sporules of the 
 middle row are better seen, and at a low focus those of the lateral 
 rows. In several specimens, especially when coloured with picric 
 acid, we can detect the prolongation of the stem into the interior 
 of the ear ; but it is not so with aniline colours, which render the 
 viscid substance (in which the sporules are suspended) opaque, 
 staining it with a weaker tint. But if, in the preparation, we sub- 
 stitute glycerine for water, the stalk, a long time afterwards, 
 reappears distinctly in all its length. 
 
 At any rate, the movements given to the preparation with the 
 thumbs show that the stalk and the " ear " form a continuous 
 
AND CONTENTS OP THE MOUTH. 119 
 
 whole ; then we observe the " ears " on the top vibrating like the 
 ears in a field of wheat. 
 
 It may be objected that the sporules are not immediately 
 grafted on the stalk or fertile filament, but are distant from it ; it 
 must, however, be taken into consideration that the distance 
 between the sporules and the internal filament is not, in fact, 
 greater than that which is constantly seen between the articula- 
 tions of the single chains of bacteria, which, as it is scientifically 
 ascertained, are generated one from another. Perhaps a connec- 
 tion exists between them, through threads or ligaments, not dis- 
 cernible by the optical power at our command.* 
 
 We cannot even suppose that the sporules are simply adhering 
 to the filament through external action (due to the lowering or 
 sinking, so to speak, of the filaments themselves into the granular 
 masses), because a simple mechanical incrustation would not 
 explain the regular straight line of the sporules in six longitudinal 
 rows, nor their perfect similarity of volume and shape. 
 
 The incrustation would bring a medley of cocci and bacteria, 
 of varied type and size, irregularly mixed up. Besides, we shall 
 observe productions of these "ears " (and even more conspicuous) 
 in the sputa, without any trace of bed or granular mass on which 
 the filaments might have become incrusted from the outside. 
 
 The length of ears is frequently considerable, especially in 
 those detached from the matrix, which at times, with their stem, 
 occupy all the visual field. The more conspicuous specimens 
 (Fig. 1 6) have been found in the pulmonary sputum, where the 
 
 * When we presented this Memoir, we promised in a note to resume the 
 study of ' ears " in detail, with more powerful objectives. Now Messrs. Bezu, 
 Hausser, and Co. have succeeded in constructing a new objective with homo- 
 geneous immersion, of l/25th inch. With this power we have been able to 
 detect, in the most evident manner, the peduncles or engrafted threads of the 
 sporules on the principal stalk. With the new grand model (No. VII.) of the 
 same opticians, furnished with an Abbe condenser of the numerical aperture, 
 I -40, by using oblique light, and by turning the stage of the microscope, or 
 substituting a dark diaphragm for the ordinary one, the disposition of the spo- 
 rules in six longitudinal lines is fully confirmed. We shall treat this point 
 again later ; but from this moment we think it is better to give the name of 
 " bunches of grapes " instead of that of " ears" (to indicate the fructifications 
 of Leptothrix), as they really are such. 
 
120 BACTERIA IN THE SPUTA 
 
 longest were one-sixth of a millimetre high, equivalent to the 
 thickness of a thin cover-glass, as in the Figs./ g of the figure. 
 
 The colouration of the ears in question with carmine, pro- 
 tracted even for weeks in a watch-glass, gave no satisfactory results. 
 With methyl violet the specimens appear less transparent ; in other 
 respects, they are like those treated with gentian violet. In Fig. 
 1 1 one may be observed, in which the ears are in the direction of 
 the visual axis. With fuchsine the most fine and brilliant coloura- 
 tions are obtained, as shown in the specimen caught in profile, 
 represented in Fig. 1 2, after being immersed for a few minutes. ' 
 
 In the uncoloured preparations, the ears are hardly or not at 
 all discernible, so that they could not be detected by him who 
 should ignore their existence ; the sporules being, in their natural 
 state, transparent and nearly invisible ; but these preparations are 
 helpful to him who knows them, in so far as they aid to detect, 
 on the outline of the mass, a larger number of ears which have 
 not been spoiled or carried away by the manipulations, or hardened 
 and consequently become brittle,. 
 
 Anyhow, the flexibility of the ears is preserved even in the 
 colourings with the solution of iodine, not acidulated, or with 
 picric acid. The sporules appear distinctly with the solution of 
 iodine (Fig. 13); but with picric acid the colouring is diffused and 
 pale, although the pale tint is useful in its turn in disclosing at 
 one time a larger number of fructifications, without rendering any 
 of them opaque. 
 
 Here we shall stop awhile to consider the great affinity of those 
 sporules through the aniline colours ; which affinity brings them so 
 closely to the tubercular bacilli as to make us suspect that the 
 single articulations of such bacilli proceed from the sporules, 
 which, having dropped, transplant themselves into the buccal epi- 
 thelia and the air-passages, and grow like beaded forms. We were 
 led to that supposition from the following fact : — In the colourings 
 with fuchsine of the patina dentaria, which last a very short time, 
 the central portion of many masses remains uncoloured.. How 
 great was our surprise in seeing, some time afterwards, in those 
 uncoloured areas, minute red lines, perfectly similar to the tuber- 
 cular bacilli ! Then those coloured lines gradually increased, and 
 we saw the summits of ears developing, as mounds, brilliantly 
 
AND CONTENTS OF THE MOUTH. 121 
 
 coloured, in a clear area. This fact is to be attributed to the 
 subsequent action of the colouring matter. 
 
 Besides the bacilli of Koch in rosaries, we have others in small 
 rods or filaments containing granules or vacuoli, as in Fig. i of the 
 work of Cornil and Babes, etc. These rods appear more clearly 
 when stained by Gram's method — viz., with double colouring, first 
 with gentian violet and then with solution of iodine. Now, the 
 fragments and slender stems of Leptothrix, in the patina dentaria, 
 behave likewise. If the solution of iodine should be used first, 
 the successive colouring with gentian violet does not succeed ; but 
 if we colour with violet, as usual, mounting the preparation when 
 fresh, and then introducing by capillarity the solution of iodine, 
 the violet colouring becomes dark blue and the small rods of 
 Leptothrix appear very clearly. 
 
 On another occasion I shall treat those questions relating to 
 the .finding and colouring of the tubercular bacilli. 
 
 Productions by Points (Male Organs ?) 
 
 I have already said that such productions or pseudo inflores- 
 cences are scanty, so that we must seek them patiently in the 
 preparation. They have, however, the advantage of showing 
 themselves clearly, even without colouring. It is absolutely neces- 
 sary to examine the patina dentaria of persons who have fasted 
 and who are not in the habit of cleaning their teeth, otherwise we 
 may not find these productions. The preparation should not be 
 pressed too much, in order to avoid the disintegration of the spe- 
 cimens. They are easily recognised with an objective. No. 8 of 
 Hartnack. The above-mentioned difficulties indicate why such 
 forms have not been considered by previous investigators. 
 
 In Fig. 14 are shown three of these productions in the natural 
 state, taken from the patina dentaria, and magnified to 690 dia- 
 meters. They are formed of one or more internal stocks and a 
 considerable number of points which surround them, like the 
 horsehairs of a cylindrical brush. In a the production rises upon 
 a multiple stalk, formed of a bundle of filaments apparently 
 broken off. The stalk itself is crowded with small points near the 
 trifurcation ; the three small divided branches bear longer points 
 
 K 
 
122 BACTERIA IN THE SPUTA 
 
 up to the top ; but the right one is also subdivided in two second- 
 ary tiny branches. In b is drawn an isolated pseudo inflorescence, 
 formed of small and medium points, like the small branches of the 
 preceding specimen. In c, on the contrary, is seen a production 
 broken on the top, formed of small, medium, and large points ; 
 the small points similar to the preceding ones; the large ones 
 totally similar to the pointed and spindle-like bacilli of type e 
 "(Fig. s), of which, for comparison's sake, we have reproduced one 
 in its natural state in d (Fig. 14), magnified to 1,250 diameters. 
 Most of the small points resemble the Virgula bacilli of type h 
 (Fig. 5); some, however, look like the spermatozoa of type K 
 (same figure), having at one end a kind of small head, according 
 to the degree to which they are magnified. 
 
 In Fig. 15 is seen a portion of a small branch, coloured with 
 the solution of iodine, magnified to 2,500 diameters. Points of 
 various size and shape are set in it, which, like the central rod, 
 contain clear spaces or uncoloured empty ones. If it were not too 
 bold a supposition, I should be inclined to suggest that the three 
 small points are not adhering to the central rod, but are fulfilling- 
 their functions upon it, and perhaps even upon the bacillus which 
 adheres to it lower down — a quid simile with the act of fecundation. 
 
 I repeat that it is not my intention to pronounce on the nature 
 of these productions by points or pseudo-inflorescences, that being 
 a subject for micetologists. I have only wished to describe facts 
 resulting from my own observations. However, I cannot help 
 noting the very extraordinary mobility exhibited by those points, 
 large and small, dropped from the stalks or branches above 
 described. 
 
 Those movements are better detected in uncoloured prepara- 
 tions, mounted in simple distilled water, in which they are pre- 
 served longer (even from day to day), especially in the minute 
 Virgula bacilli ; so that I suggest to those who will verify this part 
 of ray observations, not to use any tint at all. The stains used, 
 particularly the aniline colours, seem to have a poisonous effect 
 upon those bacilli, or at least exert a lethargic action on their 
 motility. In the largest or spindle-like bacilli of type d (Fig. 14) 
 the movements are those of translation, so that they fully justify 
 the name of Bacillus tremulus, given to them by Rappin, if that 
 
AND CONTENTS OF THE MOUTH. 128 
 
 name does not imply a separate specific entity. We notice that 
 those bacilli vibrate not only sidewise, like fishes, but flash here 
 and there with a rapid translatory motion, dashing first upon one 
 tiny island and then (after a pause) upon another, or disappearing 
 from the visual field. It is impossible not to recognise in those 
 motions the character of vital movements, either voluntary or 
 teleological ; and excluding the first hypothesis, which would lead 
 us to the animalcula of Leeuwenhoek, Ficinus, and Klencke, 
 there only remains one external purpose to this bacilli, which is 
 directed to the performance of one function. That function can 
 only be an important one, considering that those motions are the 
 highest and most persisting manifestation of life exhibited by 
 buccal microbes. Now, what more important function than that 
 of fecundation ? * 
 
 - The movements of the Virgula bacilli of type h (Fig. 5) are 
 generally undulating and serpentine ; but they have also a trans- 
 latory one, as we have suggested before. 
 
 In conclusion, we may suppose that those points, or so-called 
 bacilli, perform a function similar to that of spermatia or anthero- 
 zoids of various fungi and sea-weeds (AlGje). In such hypothesis 
 the absolute sterility of their cultures would be wholly explained, 
 being quite natural (as before suggested) that simple male organs 
 should not be able to reproduce by themselves alone. It remains 
 to be seen whether these productions by points have any relation- 
 ship with the bacillus n (Fig. 2 to the left below), a large bacillus 
 veined through its length, which contains, as in germ, slender and 
 short lineal bacilli towards the inferior end, so that we should be 
 inclined to compare it with a spermogone or antherid.\ 
 
 We have hitherto dealt with productions by points, in their 
 higher phase ; but near these we find other less developed forms, 
 
 * In the quoted article of Dowdeswell, to our surprise, we find that he 
 entertains doubts upon the nature (whether vegetal or animal) of the Cholera 
 bacillus. See Lancet, 1890, Vol. I., loc. cit., p. 1422. 
 
 tThe spindle-like and Virgula bacilli of the mouth closely resemble, in 
 shape and size, the spermatia, of the Sphcerella sentina, of the Fumago salicina, 
 of the Apiosporium ciiri, etc. The spermatia were considered, since 1877, as 
 male organs, when Cornu noticed that some of them were germinating by 
 themselves, so that their nature would be still uncertain. 
 
124 BACTERIA IN THE SPUTA 
 
 supported likewise by one or more internal stems and enveloped . 
 in a mass of very minute lineal germs, quite similar to those con- 
 tained in n (same figure). Such lineal germs, engrafted in square 
 on the stalks, resemble spindle-like, serpentine, or Virgula bacilli 
 in formation, so that the whole may be taken for the first stage of 
 the pseudo inflorescences, or productions by points, already des- 
 cribed. Owing to the difficulty of reproducing them faithfully, we 
 have omitted their graphic representation in the Plate. 
 
 Fructification by Spores (Ears) Reproduced in 
 THE Sputa. 
 
 I will now deal with the reproduction of the fructification by 
 spores (spicae) in the sputa. Hitherto we have been unable 
 to obtain cultures of Leptothrix, and microbes of the mouth in 
 general, upon the usual artificial soils ; and on this point the 
 authors, from Cornil and Babes to the more recent, are agreed.* 
 It will not be inopportune to point out a case of fructification by 
 ears, reproduced in a specimen of sputum, which induced me to 
 make the present communication. 
 
 The presence of filaments, isolated or intertwined, of Lepto- 
 thrix, in the sputa, is well known in clinical microscopy. Leyden 
 and Jaffe recognised even Leptothrix pulmonaris ; and Hunter 
 Mackenzie exhibits a conspicuous specimen taken froipi tuber- 
 cular sputum during a time of improvement.f In the sediment 
 of masses of sputa, emulsified with potash, we find not only 
 little rods and filaments, but even numerous tufts of this parasite. 
 I remember also having often found in sputa of every kind 
 freshly stained fragments which I had not known before, and 
 which were portions of ears spoiled in the preparation. 
 
 The case which gave the conspicuous specimen, delineated in 
 Fig. 1 6, was the following : — 
 
 * Cornil and Babes, op. cit., p. 135. 
 
 + Leyden and Jaff6, Deutsches Archiv far Med., II., p. 488; Hunter 
 Mackenzie, Le Crachat, etc., trans., Paris, 1888, Fig. 22. 
 
AND CONTENTS OF THE MOUTH. 125 
 
 Mr. G. F., 41 years of age, having already been twice attacked 
 by obstinate miasmatic affections, and once by an acute affection 
 of the air-passages (the nature of which he does not remember), fell 
 ill last March with very high fever (from 40-6° to 4i"2" in the 
 evening hours, falling about a degree in the morning) ; with serious 
 dyspnsea (from 45 to 50 breathings a minute), and very rapid pulse 
 (13S — 145). On the chest it was. only noticed that the right apex 
 was less sonorous, giving an indistinct respiratory murmur ; little 
 coughing, with scanty and infrequent expectoration; tumour of 
 the spleen by percussion, but probably chronic. The sensorium 
 sound ; a little insignificant epistaxis ; remarkable bilious diarrhoea. 
 Albumen, one and a-half grammes to every pint of urine, with a 
 few hyaline and granular urinary casts, and increase of ordinary 
 pigments. 
 
 In the first days there was a doubtful expectoration, which was 
 thought to proceed from the nose. An eruption of herpes labialis 
 appeared (which I remember having deceived me in a case 
 of ileo-typhus). At any rate, the diagnosis of ileo-typhus was 
 given, and three baths were prescribed which gave great relief. 
 On the seventh day the fever suddenly subsided, as generally 
 happens in pneumonia. Dyspnoea remained, only diminished by 
 the quota due to fever ; and in thirty-six hours 1 obtained a small 
 quantity of rusty sputa, a portion of which, nearly free from saliva, 
 I examined, to ascertain whether it proceeded from the air-passages. 
 
 The diagnosis -was made even clearer when, later on, crackling 
 wheezings and murmurs of friction in unison with the beatings of 
 the heart (pleuro-pericardiac friction of Wintrich) became mani- 
 fest ; these continued up to the sixteenth day of the illness. I 
 have met with this phenomenon on other occasions, in pleurisy 
 or in pleuritic granuloma, but on the left side ; here, on the con- 
 trary, it was heard on the right upon the line of the sternum, and 
 below towards the mammillary line. Consequently, the grave 
 dyspnoea had been the means of spreading the process to the 
 pleuritic coating of the pericardium, and perhaps to a limited part 
 of the pericardium itself. At any rate, the examples of a great 
 disproportion between the extension of the local and the magni- 
 tude of the general facts are not rare, especially when the apices of 
 the lungs are affected with pneumonia. 
 
126 BACTERIA IN THE SPUTA 
 
 The following were the elements found in the sputum : a fair 
 number of red blood corpuscles, both isolated and in small clusters ; 
 here and there a great gathering of ellipsoidal epithelia (so-called 
 alveolar), mostly with a coat of granules of myelin, and a few free 
 particles of the same ; many cylindrical and vibratile epithelia of 
 the air passages i the usual buccal epithelia ; a few slender and 
 short spires of Curschmann ; myriads of uncapsulated pneumo- 
 cocci, generally in vast cumuli ; small well-defined groups of 
 capsulated pneumococci, after type in Fig. 4, /; various other 
 forms of bacteria and bacilli ; filaments, and even tufts, of Zep- 
 toihrix ; groups and large cumuli of spores and sprouts of oidium 
 {Oidium lactis ?). 
 
 As it is my custom to keep the sputa for whole weeks in 
 order to repeat my investigations, on the seventh day after col- 
 lecting this sputum I detected in it fragments of fructifications, 
 which I could perfectly well recognise. I then thought of remov- 
 ing a particle of the patina of the sputum, which was adhering to 
 the internal wall of the tube, to that side on which I had always 
 inclined it, in collecting the specimens. There the layer of the 
 mucus was very thin and wet, the tube being accurately closed. 
 This mucous patina, having been adhesive and undisturbed for 
 several days, ought to have preserved nearly the same conditions 
 favourable to the fructification of Leptothrix, as seen on the 
 patina of the teeth. That layer appeared to the naked eye opaque 
 and greyish, like a very delicate mould. 
 
 The result of that research could not have been more satisfac- 
 tory, as Fig. 16 shows. It extends over two visual fields (magnified 
 850 diameters), and the colouring of ears with gentian violet 
 is very striking, the particle of the sputum having been immersed 
 for two hours in the liquid stain, then washed and mounted in dis- 
 tilled water. It will be observed at a glance that the bundle of ears 
 has been severed in two groups : in the superior one. A, all the 
 stalks are preserved; in the inferior, B, only five are visible. The 
 fructifications or part of them, as well as the missing stalks, were 
 spread here and there over the surrounding visual fields. The 
 inferior group, B, has been pressed sideways by the cover-glass, 
 and lies in profile with its base down ; the superior one. A, has 
 been pressed down vertically by the cover-glass, and is conse- 
 
AND CONTENTS OF THE MOUTH. 127 
 
 quently opened and formed into two, so that it shows five upper 
 stems with the base downwards, and two lower stem*, /and g, 
 with the base upwards.* 
 
 In the largest stalk, a, the gemmules of reserve are distinctly 
 observed, and these are still better seen in a', where a part of that 
 stalk is reproduced (magnified to 2,500 diameters) after a long 
 saturation in glycerine (see later on). In b, the ear is partially 
 scattered, so that a few spores remain in situ, and the stalk is 
 entirely visible. In d and e, cumuli of sporules are seen in those 
 points where the ears are partially scattered. Then from the 
 concussion the sporules are driven towards the top, leaving bare 
 the stalk near the base. In c, the ear is broken, and shows a clear 
 section, but without its superior part. 
 
 In the patina dentaria I never came across ears so long 
 and perfect. In fact, the better kept ears, bent down, in / and 
 g, are the longest found hitherto; as that in /measures i66/t, or one 
 sixth of a millimetre, which corresponds to one and a half the 
 thickness of a cover-glass, nine to the millimetre. The number 
 of the sporules implanted in these ears (upon six longitudinal 
 lines) may be calculated, at the lowest, to be 720. 
 
 We shall have, here, to consider two points. The first is, that 
 a similar specimen cannot be supposed to have been previously 
 formed on the teeth, then fallen in the expectoration with saliva, 
 to be found there second hand, as the fructifications do not rise 
 so high in the patina dentaria, nor could they have been so well 
 kept after the various manipulations. The circumstances under 
 which they were found incline rather to the belief that they had 
 germinated on the mucous film from filaments there deposited, 
 and were being fed from the materials of the sputum itself. 
 
 In the present case, we shall not take into consideration 
 whether the presence of red blood corpuscles may have favoured 
 the development of those ears. It is certain that they are found 
 more abundant in the patina dentaria of persons suffering from 
 tenderness of the gums, and whose teeth are, in the morning, 
 somewhat besmeared with blood. 
 
 The second point is that the specimen in question is sufficient 
 to clear up any doubt whatever about the nature of the above- 
 * In the Plate, Fig. 1 6 .5 is erroneously marked Fig. 14^. 
 
128 BACTERIA IN THE SPUTA 
 
 described ears. Taking only the specimens obtained from the 
 patina dentaria, some doubts might remain, as in it the ears are, 
 generally, embedded in the granulous lumps and the entangle- 
 ments of threads, in consequence of the pressure of the lips or 
 tongue ; and we might suppose that they are the mechanical result 
 of that trampling or superposition of minute cocci around the 
 branching filaments, so as to become incrusted with them. But 
 (apart from the consideration that the incrustation cannot explain 
 the perfect similarity of the sporules, nor their constant disposi- 
 tion in six longitudinal lines) the specimen of Fig. i6 dispels any 
 suspicion whatever, as it is there manifestly the sign of a gen- 
 uine and proper germination ; nor, on the other hand, can there 
 be found a particle of resting place on which the stems might 
 have become incrusted. 
 
 In support of this view there is the fact that the ears acquire, 
 by colouring with aniline, such a solidity that they can hardly be 
 dissevered. The above-mentioned specimen, which, through the 
 accidental crushing of the preparation by a wrong turn of the fine 
 adjustment, was hard-pressed down, exhibits a proof of it. The 
 ear drawn in g was abruptly removed towards f, and made into a 
 spire, but no spores were dropped by the shock, and the ear has 
 since gradually resumed its former place. Now this fact could 
 not be explained on the hypothesis of a simple mechanical 
 incrustation. 
 
 This preparation has been kept in glycerine, which was after- 
 wards substituted for the aqueous medium. The tint, especially of 
 the pneumococci without halo, is very much faded, owing to the 
 diffusion caused by glycerine. The ears, after two months of 
 saturation, exhibited a partial decolourisation and withered spores, 
 although the stalks maintained a brilliant colour. The viscid 
 matter has become paler and more transparent, so that the stalks 
 can be detected fully in their whole length ; and inside can be 
 seen not only the gemmules, but traces of small knobs, shown in 
 a'. By turning the micrometer screw I could even see, although 
 _ interruptedly, a few sporules belonging to the posterior series or 
 those lying on the slide. 
 
 In conclusion, having thoroughly investigated this preparation, 
 it seems to be absolutely impossible to dispute the fructification of 
 
AND CONTENTS OF THE MOUTH. 129 
 
 those forms of Leptothrix and the fertility of their stems. This 
 specimen is at the disposal of an)'one who wishes to examine it. 
 
 We shall, by-and by, deal with the fructification obtained from 
 urinary mucus. 
 
 With regard to the oidium forms found in this sputum, I will 
 say two words concerning their fructifications. I have dealt with 
 this argument formerly (see previous Memoir), and spoken about 
 cultures upon peels of lemons and various other nutrient media ; 
 but I have found it more e.x:pedient and satisfactory to let the 
 fungi of the sputum germinate in the sputum itself. 
 
 The better soil for cultures is that in which spores and branch- 
 ing filaments carry out their immersed vegetation. In order that 
 aerial vegetation should take place, two conditions are required : 
 the first is that the materials of culture (in this case, the sputum) 
 should be moderately dry on the surface ; and the second is that 
 the under layer, or the lowest stratum of the sputum, should be 
 kept wet in order to feed the vegetation. Now, both conditions 
 are fulfilled by placing a part of the sputum, impregnated with 
 spores and sprouts, on the bottom of a wine-glass slightly hollow, 
 so that the sputum should sufficiently spread and rise to four or 
 five millimetres. The wine-glass and its cover should be previ- 
 ously cleansed with sulphuric acid, and washed out with alcohol. 
 Through the wide opening of the glass the fructifications can be 
 easily observed and scraped out for microscopical examination.. 
 Generally, fructification is completed on the fourth day, by keep- 
 ing the glass well sheltered in the dark. 
 
 The sputum, impregnated with oidium forms, exhibited on the 
 fourth day a simple fructification of Penicillium glaucum. Natu- 
 rally, we do not intend by this to discuss the question brought 
 forward by HalHer — i.e., whether the oidium forms be one phase of 
 the Penicillium and Mucor kinds. 
 
 Dissemination of the above-described Microbes on 
 THE Posterior Organs 
 
 Considering now the extended vegetation and fructification of 
 Leptothrix upon the dental surface and superposed layers; its 
 abundant germination on the tongue, its constant presence in the 
 epithelia of the cavities of the mouth and pharynx, its very active 
 
130 BACTERIA m THE SPUTA 
 
 multiplication in saliva, the countless swarms of analogical forms, 
 quiescent or reproductive, in the whole wide surface of the nasal 
 cavities, and perhaps of its appendages ; considering also the 
 extraordinary fertility of this parasite, and its multiplication not 
 only through the fission of cocci and bacteria, but through fructi- 
 fication and budding within the filaments, we may form an approx- 
 imate idea of the mass of germs or scattered elements which are 
 invading the digestive, aerial, and lachr5rmal passages. 
 
 According to my calculations, not less than from two to three 
 hundred trillions of germs or separated elements are generally - 
 present in the mouth and nose, and liable to disseminate the 
 species, at every minute, into the other parts. 
 
 In fact, comparing the single sporules of the ears with the 
 dumb-bell bacteria of types a, b, c, d (Fig. 2), we should have to 
 place nearly eight sporules in two rows upon the surface of a single 
 bacterium in order to cover it. The bacterium is twice as thick as 
 the sporule ; consequently, the volume of one bacterium will be, 
 at least, equivalent to that of sixteen minute sporules. Now, in 
 our first Memoir, we have demonstrated that about 25 milliards 
 of bacteria of types a, b, c, d, go to form a centigramme in 
 weight ; then, for every centigramme of sporules, we shall 
 have to multiply the 25 milliards by 16, which will give 400 
 milliards of sporules for each centigramme. Calculating now 
 the considerable number of the dropped sporules, that of 
 the gemmules of reserve in the filaments, that of even the 
 most minute granules which constitute the bed of the branch- 
 ing threads and of the clods of Leptothrix; calculating the very 
 small lineal germs grafted on the young productions by points, 
 we can safely maintain that their extreme minuteness compensates 
 for the larger volume of bacteria enclosed in the older filaments, 
 that of spindle-like, the comma, and the serpentine bacilli, as well 
 as of spirilla and cocci disseminated in every part. Thus, holding 
 the volume of the sporule as the unity of measure, taking the 
 whole patina of the thirty-two teeth as equivalent, at least, 
 to a gramme, we shall have the number of the elements and 
 germs living on the 'dental patina equal to the product of 400 
 milliards (in about a centigramme), x 100, viz., 40 trillions. 
 To these figures is to be added the huge mass of germs and 
 
AND CONTENTS OF THE MOUTH. 131 
 
 elements spread in the saliva, of which a third part may be con- 
 sidered to be formed of microbes : ut tota aqua vivere videatur, as, 
 in his time, Leeuvvenhoek wrote. The quantity of saliva which 
 continually moistens the mouth is not less than eight or ten 
 grammes ; therefore, we have three more grammes of germs and 
 elements, bringing the total up to 120 trillions. 
 
 But we will not further trouble the reader by calculating the 
 other germs and elements lodged on the tongue, in the mucus of 
 the mouth and pharynx, and in the nasal cavities ; from which it 
 would result that the figures given above are not at all exaggerated. 
 
 It is to be noted that we started with an average of 25 milliards 
 of bacteria per centigramme ; a figure twelve times under the cal- 
 culations of Naegeli— i.e., 30 milliards per milligramme, which 
 would make the mass of germs and elements of nose and mouth 
 upwards of 2 or j quadrillions. 
 
 Besides, we must consider that Leptothrix lodges, as we have 
 demonstrated, even in the mouth of domestic animals. Now, the 
 mucus, saliva, breath, urine, faeces of these animals, jointly with 
 those of men, constitute, in their whole, an immense preserve. I 
 am led to believe that a great many germs and elements of Lepto- 
 thrix pass into the faecal matters alive. There is also incalculable 
 diffusion in the air, waters, and soil, especially in populous towns. 
 What wonder, then, that Perroncito has found such a large number 
 of bacteria in the dust of the streets ? We may, rather, contend 
 that such a dissemination of buccal microbes constitutes a sort of 
 cloud, which hardly permits external germs to penetrate. 
 
 But let us return to the dissemination of Leptothrix in the 
 internal organs of the human economy. 
 
 With regard to the digestive passages, the number of elements 
 carried there by the saliva must be fabulous, considering that in 
 the smallest drop myriads of them are to be found. Nor is it 
 presumable that their descent into the stomach should prove 
 
 inactive. 
 
 The view that they are partakers in the transformation of ali- 
 ments is not new ; it goes as far back as Hallier, who considered 
 them necessary to the transformation, especially of starchy sub- 
 stances, not only in the stomach, but even in the mouth itself.* 
 
 * Richter, Monograph quoted in the BibUography, Part I., Section B 2. 
 
132 BACTERIA IN THE SPUTA 
 
 B^champ compared the transformation of saccharine in the pre- 
 sence of its microzymas with that of the fsecula through diastases.*' 
 Warkmann held the same action even for starch, and he conceived 
 that from the bacteria was secreted a true diastatic yeast, destined 
 to transform starch into glucose, although unapt to peptonise the 
 albumenoids.t But, later on, others recognised in buccal bacteria 
 even such a peptonising action, and Miller holds it to be equal to 
 that of the pepsine itself, even without the co-operation of acids.J 
 
 A considerable number of the same germs and elements of the 
 mouth, as well of the nose, is cast into the air-passages at every 
 breath, as the daily observation of the normal sputa shows (see 
 preceding Memoir). Those germs and elements thrive and multi- 
 ply in the mucus of the bronchi, trachea, and larynx, as the 
 fructification in the pulmonitic sputum, just described, shows ; it 
 is evidently a soil fit for the vegetation of Leptothrix. 
 
 Shingleton-Smith,§ in the quoted paper on Contagium Vivum, 
 considers, first, the property that the moistened mucous membrane 
 has of holding bacteria. It does not let them out with the breath, 
 but through the ciliary action of the epithelia, which throws them 
 out with the mucus. This happens with pathogenic bacteria, but, 
 a fortiori, it must be so with normal bacteria. The same author- 
 quotes the experiments of Tyndal, who found the residual air from 
 the lungs absolutely free from particles which would reflect the 
 electric beam. He quotes as well the observations of Lister and 
 others, who maintained that there were no bacteria in the pus of 
 Empyema with Pneumothorax, the opening on the surface of 
 the lung being very small. This must indicate that the microbes 
 
 *B6champ, Les Microzymas et les Zymases, Arch, de Physiol., 1883. 
 The same, La Salive, la Slalozymase, et les Organismes Buccaux, etc. {vide 
 Bibliography), 1883. 
 
 t Warkmann, Untersuch, iiber das diastatische Ferment der Bacterien 
 ( Zeitschrift far Phys. Chemie, Bd. vi., 1883J. 
 
 % See Bufalini, Of the Peptonising Action of Bacteria ( Giornale internaz. 
 delle Sc. Mediche, 1883). Miller, Ueber Gahrungsvorgange im Verdauungs- 
 tractus und die dabei betheiligten Sfaltpilze (Detit. m. Woch., 1885, No. 49). 
 The same, Die Mikroorganisme7i der Mundhohle{ste'5\h\\ogt3.^hy), pp. 77 — 90, 
 where he specially deals with the fermentative action of buccal microbes upon 
 hydro-carbons, albumenoids, and fats. 
 
 % Journal of Microscopy and Natural Science, iSgo, p. 34. 
 
AND CONTENTS OF THE MOUTH. 133 
 
 are kept back by the mucus wetting the larger air-tubes, where the 
 protecting barrier of the epithelia exists; but they cannot reach 
 the slender bronchi or alveoli where the epithelium is evanescent 
 or at all wanting. Thus, the microbes, having reached the border 
 of the smallest bronchi, by means of the ciliary action, they are 
 thrust back towards the superior passages, and carried at last, by 
 the sputum, into the normal or pathological mucus. 
 
 But if this happens in normal conditions, it cannot be so in 
 certain pathologic conditions, as in pneumonia, pleurisy, and 
 phthisis. The elements or parasitic germs, in these cases, not only 
 reach the most minute bronchi, but from there they diffuse them- 
 selves even to the remotest parts; this may proceed from the 
 insufficiency of the ciliary action, or from an extraordinary con- 
 course of parasitic elements overcoming it, or from the microbes 
 along a denudated tract of mucous membrane. 
 
 Forster and Graefe* were the first to write upon the vegetation 
 of filaments or branching threads of Leptothrix on the conjunc- 
 tival or lachrymal ducts ; Bizozzero draws a specimen in PI. V., 
 Fig. 52, of the quoted work. This has been reproduced in our 
 Fig. 6, d, and it is natural to suppose that the relative germs origi- 
 nate from the nose. 
 
 If now, from the buccal and nasal microbes, we pass to con- 
 sider those of the external genito-urinary passages, we shall find 
 the same disseminations; but with this difference, that they will not 
 be found so frequently and in such abundance. This depends 
 upon two reasons : — The first is the relative scarcity of parasitic 
 vegetation, especially on the balano-preputial mucus (compared 
 with that of the mouth and nose together) ; the second is the 
 want of a propelling force, sufficiently vigorous and repeated, 
 capable of pushing, at least in normal conditions, the elements or 
 the germs of the parasites in the urethra or in the bladder. At 
 any rate, we must admit a penetration, on a small scale, by looking 
 at the epithelia of the inferior urinary passages. Such epithelia, 
 even in the most normal and freshly-discharged urine, are found 
 impregnated with bacteria to such a degree that they cannot be 
 attributed to consecutive pollution (viz., after the urine has been 
 
 * Forster, Arch, fur Ophtalm., t. XV., page 310. Graefe, Ueber Leptoth- 
 rix in den Thranenrohrchen (Arch, cit., t. xvi., p. 324). 
 
134 BACTERIA IN THE SPUTA 
 
 passed) ; but they evidently denote their invasion in situ in the 
 bladder or the urethra. Of course, the invasion has happened by 
 a slow, progressive motion of the relative elements or germs, 
 either along the mucus coating, or from one epithelian scale to 
 another, and so forth. 
 
 In certain urines, round the small flakes or spires of mucus of 
 the male urethra, clods of Leptothrix are met, at times, so large as 
 to occupy the whole visual field of an objective (Nos. 7 or 8 of 
 Hartnack). Such clods are so closely similar to the young clods 
 of the patina dentaria, that the most experienced investigator 
 would be unable to distinguish one from another. / 
 
 Lately, I observed a case of this kind in the urine of a patient 
 affected with chronic pyelitis, having already been subject to 
 several attacks of Blennorrhoea. I placed two of these small 
 flakes in a closed tube with a little of the urine, and for six days I 
 kept them adhering to the internal wall, wetting them gently once 
 or twice a day by inclining the tube. On the sixth day the mate- 
 rial of these small flakes, prepared and stained with gentian violet, 
 like the patina dentaria, exhibited exactly all the forms of Lep- 
 tothrix biiccalis, with the exception of the productions by points 
 (perhaps spoiled in the preparation). Undoubtedly, it contained 
 countless bacilli, both comma and spindle-like, rather small, and 
 in brisk activity ; even a fair number of small ears was to be found 
 there. Briefly, all forms of bacteria and bacilli, chain-like fila- 
 ments, and btmdles, described in Leptothrix buccalis (except the 
 Jodococcus vaginatus) were likewise there ; and the staining also 
 showed that not even the spirilla or spirochmte, mostly in motion, 
 were wanting. 
 
 Between these forms of Leptothrix and those of the mouth, 
 the only difference is that the filaments, bundles, and chains are 
 shorter and the spirilla extremely slender, but longer. In all other 
 particulars they were perfectly identical.* 
 
 * Our present remarks about the multiplicity of the microbes of Blennor- 
 rhoea and the presence of spirilla in urethral secretion have been confirmed by 
 the research of Legrain, On Blenorrhaa microbes. That author found in the flow 
 of gonorrhoea 12 different forms of cocci, 3 distinct forms of bacilli, and i spi- 
 rillicfortn : total, 16 forms of microbes ( Archiv f. Dermatologie u. Syphilis, 
 1890). Stroganoff has recently dealt with the bacteria of the vagina and cervi- 
 cal canal.— {Modem Medicine and Bacteriological World, Oct., 1893, p. 258. > 
 
AND CONTENTS OF THE MOUTH. 135 
 
 It would be useful to persevere in this kind of research, and I 
 propose to resume it on the first opportunity, in order to prove the 
 identity of Leptothrix huccalis with praputialis, and then that of 
 bacteria and bacilli of the air-passages with those of the genito- 
 urinary organs. 
 
 We have an argument by analogy for the arrival of the elements 
 or germs of Leptothrix in the urethra, in the introduction of more 
 bulky germs, like those of superior fungi, probably of the genus 
 Penidllium, either in the urethra itself or in the bladder, of which 
 we have already given an instance in our article upon fungi of the 
 male urethra. 
 
 Naturally, the removal, insignificant in healthy conditions, 
 increases in the morbific ones, and that is the reason of the 
 gonococci in blennorrhcea ; but we have observed beforehand 
 that in blennogenous pus the most common form of bacteria is 
 not that of gonococci or curved diplococci of type g, /, p (Fig. 2), 
 but that of common diplococci with round heads, sometimes 
 surrounded by a halo, either within or outside the epithelia or the 
 corpuscles of pus. I have already treated this argument in the 
 other paper upon a case of carcinoma of the bladder, in which 
 the urine sho'wed a considerable number of curved diplococci or 
 gonococci, indeperidently of any blennogenous contagion whatever. 
 
 In the Memoir on Whooping-cough, and in the first section 
 of this work, I have touched upon the presence of such curved 
 diplococci or gonococci in the sputa or saliva ; as well as upon the 
 singular discovery of true diplococci in a halo, morphologically 
 identical with the so-called pneumococci, in the spermatic fluid 
 recently discharged through illness. 
 
 I think that the surprise or incredulity which at first may have 
 been aroused from the present observations will cease, when it is 
 considered that the incentive of all these varied disseminations is 
 always the same, i.e., that the gonococcus as well as the pneumo- 
 coccus or the bacillus of Koch proceed, in great probability, from 
 small seeds of the same genus ; from the parasite that lives nor- 
 mally at the entrance to the digestive and air passages, at the 
 egress of the lachrymal as well as the genito-urinary passages. 
 What wonder, then, if the pneumococcus, towed, so to speak, 
 along the urethra, is found in the spermatic fluid ; and the gono- 
 
136 BACTERIA IN THE SPUTA 
 
 COCCUS, in its turn, is found in sputa and saliva ; and that the 
 bacillus of Koch is met with, at times, even in the buccal epithelia 
 of a healthy man ? 
 
 For the rest, anyone is in a position to verify the exactness of 
 our observations upon these varied points.* Now from the 
 exhibited facts we must infer that probably the same bacteria or 
 bacilli considered pathogenic (in the affections of the genito- 
 urinary or air passages) are, in reality, only so many disseminations 
 of germs or elements of buccal microbes or balano-prceputialis, 
 from which, morphologically, their varied types do not differ at 
 all ; or, at least, that, before admitting the existence of this or that 
 pathogenic bacterium in the above-mentioned passages, it is neces- 
 sary to demonstrate, with clear and conclusive proofs, that such 
 bacilli, declared pathogenic, are not derivations from the normal 
 preserve. And this objection does hold good, not only in the 
 hypothesis we have set up of the oneness, or duality at most, of 
 the parasitic species of the mouth, but even in the hypothesis now 
 prevailing of their plurality. We contend that the more species 
 there are in the nasal crypts or the cavities of the mouth, the 
 more difficult will it be to exclude their co-operation in generating 
 bacteria reputed pathogenic. 
 
 Everything, indeed, leads us to believe that this inexhaustible 
 preserve of normal microbes, placed by nature at the entrance of 
 the digestive and air passages, may have a defensive mission 
 against the intrusion of micro-organisms from without ; may, in 
 other words, constitute a true excluding vegetation ; a barrier to 
 the effect of preventing foreign germs penetrating and thriving in 
 the adjacent organs. 
 
 Nature has imparted to Leptothrix buccalis such a power of 
 tenacious resistance to foreign agents that the elements of this 
 fungus upon the human teeth are not destroyed for ages, as it is 
 
 We have lately noticed that our views are confirmed in an observation of 
 Bordoni Uffreduzzi and of Gradenigo, who found " in the pus of the left ear 
 (in a case of ottorhoea) numerous diplococci of biscuit shape, some isolated, 
 others joined in tetrahedrons, partly free and partly contained in cellules, in 
 the whole similar to gonococci. " The authors quoted maintain that the various 
 microbes of ottorhoea (in which the lanceolatus diplococcus predominates) 
 originate even from the saliva. SulV etiologia delfotite media. Archivio per le 
 scienze mediche, Vol. xiv., 1890, pages 276 and 278. 
 
AND CONTENTS OF THE MOUTH. 137. 
 
 proved from the dental tartar of the Egyptian mummies, in which 
 its filaments have been found intact by Zopf and Miller, by means 
 of dissolving the calcareous salts with acids.'" 
 
 We would like to make further considerations upon certain 
 points of the present Bacteriological doctrines ; but, in order to 
 keep as far as possible within the clinical area to which our re- 
 searches are directed, we come to the final conclusion. 
 
 RECAPITULATION, 
 
 In summing up what has been exhibited in the Bacteriological 
 part of the preceding Memoir and the present one, we may con- 
 clude as follows : — 
 
 I. — Amidst all forms or types of bacteria or bacilli to be found 
 in the sputa, normal or pathological, there exist points of transi- 
 tion, manifesting their polymorphism, and the gradual passage from 
 one type to the other. 
 
 II. — Of the types in question, there is not one which cannot 
 be found even in the contents of the mouth, or in the nasal 
 mucus, and the balano-praeputialis patina. They do not morpho- 
 logically differ from each other; therefore, it is generally main- 
 tained that the bacteria and bacilli found in sputa, in normal 
 conditions, are simply secondary disseminations of the buccal or 
 nasal microbes. 
 
 III. — But when we deal with bacteria, rightly or wrongly 
 reputed pathogenic— (as, for instance, the pneumococcus or the 
 bacillus of Koch)— it is another matter. We cannot grant their 
 buccal origin, setting up, instead, the hypothesis of their having a 
 specific origin from without. 
 
 Nevertheless, even these types cannot be morphologically dis- 
 tinguished from their corresponding types of the contents of the 
 mouth or of the nasal mucus. We should have, at least, to 
 demonstrate, in a positive manner, that the former do not proceed 
 from the latter, although those same microbes are (like the other 
 buccal microbes) thrown by swarms into the air-passages. But that 
 demonstration has not been given ; nay, the only notion upheld 
 
 * Zopf, Die Spaltpilze, Breslau, 1883, page 80. Miller, Prehistoric Teeth 
 {Independ. Practitioner, 1884). 
 
138 BACTERIA IN THE SPUTA 
 
 from their resisting the decolourising process has, in some instances, 
 been proved wrong. 
 
 IV. —Consequently, there only remain, in support of the spe- 
 cificity of such bacteria, the results, more or less controverted, of 
 the methods of culture. and inoculation. We shall speak by-and- 
 by about the methods of culture, having already touched on those 
 of inoculation ; but leaving, for the present, that question on one 
 side, we wish to deny that the hypothesis of the speciality of 
 bacteria, found in pathological sputa, is at all supported by the 
 general clinical facts, or even by the daily microscopical obser- 
 vation of the sputa. 
 
 V. — The congeries of the buccal and- nasal microbes, put 
 together, may be summed up to two hundred or three hundred 
 trillions of germs and elemAits in continual prolification. This 
 vegetation, either through its diffusion — (at least, in many species 
 of domestic animals) — or its constancy and abundance, assumes 
 the character of a true excluding vegetation, placed by nature at 
 the ingress of the digestive and air passages, to aid the former in 
 the digestion, and to defend the latter against the micro-organisms 
 of the external world. An identical vegetation 'adorns the egress 
 of the genito-urinary passages. Thence, it is riot surprising if 
 bacteria reputed specific of the air-passages are found even in the 
 products of the genito-urinary passages, and vice versa; if, for 
 instance, incapsulated diplococci (pneumococci) are found in the 
 urethral mucus or in the spermatic fluid, and curved diplococci 
 {gonococci of Neisser, reputed specific forms of blennogenous virus) 
 are found, in their turn, in the sputa or in the middle ear. 
 
 VI. — Whatever may be the physiological importance of Lep- 
 tothrix buccalis, it results from our observations that its degree of 
 organisation is far superior to what has been reputed hitherto. 
 Leptothrix does not only live as bacterium, bacillus, or filament ; 
 but it possesses real organs of reproduction by which it would" 
 resemble fungi and dioecious algce, with distinct sexes upon different 
 filaments or individuals. Its fertile filaments are at times engrafted, 
 with two or three roots, upon clods or firm substrata^ and end in a 
 fructification. The ears constituting these fructifications, as long (in 
 pulmonitic sputum) as i/6th of a millimetre, are formed of many 
 very minute sporules (so much so that 400 milliards of them hardly 
 
AND CONTENTS OF THE MOUTH. 139 
 
 would weigh a centigramme) ; and the small sporules, taking a 
 bright colour with aniline, are disposed in six longitudinal series, 
 making, 'in some instances, a total of 720 for each ear. They are 
 linked together and fastened to the stalk by means of an amor- 
 phous substance difficult to be coloured.* However, other fila- 
 ments, less numerous than these, at times multiple, and lastly, 
 branching off, bear certain productions by points, or pseudo inflo- 
 rescences, formed of spindle-like, snake-like, or comma bacilli 
 (Spirillum sputigenum), destined from all appearance, through 
 their lively activity, to the function of conjugation. Finally, there 
 are gemmules in reserve, which (together with the nmltiplication 
 of the proper sporules) are destined to diffuse the species in the 
 unstable substrata or in the products and in the liquid secretions. 
 
 The fructification by ears can be reproduced, even in the sputa 
 and in certain small flakes of the urethral mucus. 
 
 VII. — Of the six primary species of fungi of the mouth, lately 
 described by Miller, there would, in fact, exist only one — the Lep- 
 tothrix buccalis of Robin {Leptothrix innominata of Miller), or, at 
 most, a second one — the Spirillum {Spirochezte dentium of Miller). 
 The other four types would represent, if we are not mistaken, only 
 phases or disintegrated particles of the microphyte — viz.. Bacillus 
 buccalis maximus and Lepiothrix buccalis maxima, fragments of the 
 stumps that form the inferior layer of vegetation; \h& Jodococcus 
 vaginatus series of special sheaths of bacteria proceeding from 
 certain gemmules of reserve enclosed in the filaments ; the Spiril- 
 lum sputigenum (comma bacilli) with our spindle-like and serpen- 
 tine ; appendages detached from the pseudo inflorescences, and 
 probably male organs. 
 
 All these particles or articulations cut from the mother plant 
 (except the last — viz., copulative filaments) multiply by them- 
 selves, in various ways, according to the condition of the nutrient 
 substratum, in the liquid menstrua or on firm soil. 
 
 VIII. — The study of such vegetable forms, in the contents of 
 the mouth as well as in sputa (especially of the fructification by 
 ears), requires special rules and care and proper optical means, 
 
 * Owing to the connecting of the sporules to the central stalk, by means of 
 peduncles or engrafting threads, visible with a new objective (i/25th in. obec- 
 tive), see a former Note. 
 
140 BACTERIA IN THE SPUTA 
 
 without which it would be difficult to verify the facts, even for the 
 most experienced investigators. The productions by points are 
 fairly well detected, even with the ordinary objectives ; but special 
 care must be taken in collecting and preparing the patina dentaria. 
 
 We do not know whether the specific oneness of the above 
 forms (by ears and by points) will be well received by competent 
 observers, as well as all the buccal microbes reunited in a single 
 plant. But, were even two or more of the vegetable species in 
 question, the succes.sive dissemination of their germs and elements 
 in the air-passages would not at all be invalidated. 
 
 IX. — The most minute form of cocci, scattered or in a line, 
 found in the contents of the mouth, in the omonimous epithelia, or 
 in the sputa, are, in our opinion, nothing but sporules dropped from 
 the fructifications by ears, being first disseminated in the mouth and 
 then thrown into the respiratory passages. The baciUi of Koch (we 
 are speaking of bead-like bacilli) might be only these same sporules 
 disposed in series,-and thus germinating in the tubercular products 
 and elsewhere. Undoubtedly, the sporules, still attached to the 
 fructifications, or fallen near them, fix strongly the aniline colours ; 
 but it remains to be seen whether, in the sputa of consumptives, 
 their resistance to decolourising means is original or simply 
 acquired in the fresh nutrient substratum received from the tuber- 
 cular lesions. At any rate, that resistance is always relative, and is 
 of doubtful value in the diagnosis, and is sometimes inferior to 
 that of other bacteria, as we have demonstrated before. 
 
 The forms of the bacilli of Koch being, on the contrary, in 
 small rods containing granules or vacuoles, with double staining 
 (gentian violet and solution of iodine) behave like the analogous 
 articulations of Leptothrix. But upon these and other not less 
 important points, about the clinical study of the tubercular sputa, 
 we shall have to deal on another occasion. 
 
 X.^However, if our observations on the morphology and 
 biology of Leptothrix are correct ; if all, or nearly all, forms of 
 bacteria and bacilli to be found in the sputa, are nothing \)\xX. par- 
 ticles or various organs of a single plant, everyone can see the 
 extent of the actual methods of culture (at least of the bacteria of 
 the air-passages). 
 
 And, to be sure, one thing it is to identify or qualify a fungus 
 
AND CONTENTS OF THE MOUTH. 141 
 
 on the ground of its fructification ; another, on the ground of the 
 form of cultures, immersed or creeping, of its single particles, or 
 the modifications brought about by these in the various nutrient 
 substrata. One bacterium or bacillus will fluidify gelatine or 
 elaborate certain principles (even poisons) in a manner totally 
 different from another bacterium or bacillus, without, however, 
 considering the two forms as two organisms or different species, 
 when such forms are for us simply the result of organs or particles 
 of the same plant, destined to attain dissimilar aims, and which 
 may possess the most different qualities. 
 
 XL — Under such conditions, the name of bacterium or bacil- 
 lus can no longer be considered as synonymous of micro-organism. 
 We may speak of bacteria and bacilli, if by those names we mean 
 particles or articulations detached from the mother plant ; and in 
 such a case the generic noun of microbes may be even applied. 
 But the word micro-organism applies to the whole microphyte of 
 which bacteria and bacilli are only single scattered particles, and 
 which do not constitute by themselves complete organisms, although 
 they mostly possess the faculty of multiplying on their own account. 
 Such name does not suit at all the single particles, nor, of course, 
 the single bacteria or bacilli." 
 
 I cannot conclude this paper without a short statement. 
 
 Perhaps some of my views will appear too bold, but my only 
 intention was to submit them to the judgment of scientific observ- 
 
 * In confirmation of our views, we shall quote the opinion of Klein, who 
 lately presented to the Royal Society of London the photographs of a culture of 
 the tubercular bacillus, which appeared with distinct branches like a mycelium 
 'fungus. He inferred that at least a few schiwphytes are really oraXy forms of 
 development or transitory phases of stiperior organisms. The opinion of Dowdes- 
 well is even more explicit. " It is clear " (writes the latter, on the subject of 
 comma bacilli) "that either these microbes are not normal schirophytes, or if so 
 they do not represent an independent group (as it has been observed by the last 
 writers on Bacteriology), but are simple phases of evolution of some superior 
 organisms, or perhaps they are only simple organs of other organisms 
 {Lancet, 1890, Vol. I., loc. cit.,\:>. 1422). Lately, Sheridan Delepine, studying 
 the development of bacteria in its cultures in interlamellar films (between 
 the cover-glass and the slide) has come to that conclusion — viz., of the branch- 
 ing off in many bacilli, by means of defined filaments (A New Method, Inter- 
 lamellar Films, of studjdng the development of Micro-Organisms, etc., in the 
 International Journal of Microscopy, Nov., 1 89 1, p. 343). 
 
142 BACTERIA IN THE SPUTA 
 
 ers. I shall only notice that my opposition to certain points of the 
 present Bacteriological doctrines is more apparent than real. I am 
 of opinion that, by pursuing the present method of specifying, in 
 the classification of bacteria, we must more and more multiply their 
 species to such an extent as to hinder the further progress of these 
 studies. On the other hand, should my observations be correct, 
 if the forms of bacteria of the air and genito-urinary passages are 
 reduced to a single species, or if, consequently, even the other very 
 varied types of bacteria (either not at all patjiogenic or pathogenic 
 of other parts) might be gradually reduced to defined species of 
 micro-organisms, according to the natural phases of their full devel- 
 opment ; this work upon their arrangement and simplification may 
 aid in the researches of experimental pathology. In other words, 
 we shall be able to recognise the difference between the botanical 
 and the pathogenic entity of bacteria ; that two or more bacteria, 
 totally different in their own pathogenic action, may proceed from 
 a single micro-organism, and that a bacterium, not pathogenic, 
 may belong, in its turn, to the same micro-organism from which a 
 pathogenic bacterium proceeds. 
 
 F. ViCENTINI, 
 
 Chieti, June, i8go. Corresponding Member. 
 
 Bibliography. 
 
 I. — Arndt, Beobactungen an Spirochaete denticola (Arch.' fiir 
 
 path. Anat. und Phys. und d. klinische Med. t. lxxiv. j 880). 
 2. — Baume, Odontologische Forschungen, II. Of the same, see 
 
 also the " Elements of Dentistry '' (Lehrbuch der Zahn- 
 
 heilkunde, 3. Auflage, Leipzig, 1890). 
 3. — Beale, The Microscope in Medicine, London, 1878, p. 490. 
 4. — Bechamp. La salive la sialozymase et les organismes buccaux 
 
 chez I'homme (Archives de physiologic, 3 serie, t. I., 1883, 
 
 P- 47)- 
 5. — Bizzozero, Manuale di Microscopia CHnica, Milan, 1882, 
 
 pp. 103— 1 04 and p. 153. 
 6. — Chevalier. L'^tudiant micrographe. Traits theorique et 
 
 pratique du microscope, etc. Paris, 1882, pp. 393 — 94. 
 
AND CONTENTS OF THE- MOtJTH. 143 
 
 7.— Cornil et Babes. Les bactdries et leur role dans I'anat. et 
 
 I'hist. patholog. Paris, 1885, pp. 28 and 135. 
 8. Decker und Seifert. Ueber Mycosis leptothrica pharyngis , 
 (Miinchen medicin. Wochenschrift. xxxv. 4., p. 67, 1888, and 
 Sitz. Ber. der physik. med. Ges. zu Wiirzburg, 2, p. 26). 
 9.— Dowdeswell. Note on the Morphology of the Cholera com- 
 ma bacillus (Lancet, 1890, Vol. I., pp. 1419—23). 
 10.— Ewart and Geddes, Life History of Spirillum (Proceedings 
 
 o'f the Royal Society, xxvii., 1878, p. 484.) 
 II.— Fliigge. I microorganism! ed etiologia delle mallattie 
 
 infettive. It. trans. Naples, 1889. 
 12. — Forster. Arch, fiir Ophtalm., xv., p. 318. 
 
 1 3- — Frey. Manuale di Tecnica Microscopica. It. trans. 
 Naples, 1873, p. 278. 
 
 14- — Graefe. Ueber Leptothrix in der Tranenrohrchen (Arch, fiir 
 Ophtalm., t. XVI.). 
 
 15- — Hallier. Die pflanzlichen Parasiten, etc., Leipzig, 1866. 
 
 16. — Jacksch. Manuel de diagnostic des malad. intdr. par les 
 
 methodes bactdriologiques, etc. (translated from the German), 
 
 Paris, 1888, pp. 50 — 57, and p. 71. 
 17. — Kiitzing. Linnaea, 1883. 
 i8.^Leber. Quarterly Journal of Microscopical Science, 1874. 
 
 19. — Leber und Rottenstein. Untersuchungen iiber die Caries 
 
 der Zahne, Berhn, 1867. 
 20. — Lewis. A Memorandum on the Comma-shaped Bacillus 
 
 alleged to be the cause of Cholera (Lancet, 1884, Vol. II., 
 
 P- 513)- 
 21. — Miller. Der Einfluss der Mikroorganismen auf die Caries 
 
 der Menschlichen Zahne (Arch, fiir exp. Pathologic, xvi., 
 
 1882). 
 22. ■ Ueber einen Zahnspaltpilz, Leptothrix gigantea 
 
 (Berichte der Deutschen Botanischen Gesellschaft, Berliuj, 
 
 [883, Heft 5, p. 221). 
 23. Zur Kenntniss der Bacterien der Mundhohle, 
 
 1884. 
 
144 BACTKRIA IN THE SPUTA 
 
 24.— Miller. Biological Studies on the Fungi of the Human 
 
 Mouth (Indep. Practitioner, 1885, pp. 227 — 83). 
 25. Beitrage zur Kenntniss der Mundpilze (Deutsche 
 
 Mediciii. Wochenschrift, xiv. 30, 1888). 
 26. — Die Mikroorganismen der Mundhohle. Die 
 
 ortlichen und allgemeinen Erkrankungen, etc., Leipzig, 1889. 
 27. — Perroncito. I parassiti dell' uomo e degli animali utili. 
 
 Milan, 1882, p. 56. 
 
 28. II carbonchio, 1885. 
 
 29. — Rappin. Les bactdries de la bouche a I'^tat normal et dans 
 
 la fibvre typhoide. Paris, 188 1. 
 30. — Rasmussen. Ueber die Cultur von Mikroorganismen vom 
 
 Speichel gesunder Menschen, Kopenhagen, 1883. 
 31. — Richter. Die Neuern Kenntnisse von den krankmachenden 
 
 Schmarotzerpilzen, nebst phytophysiologischen Vorbegriffen 
 
 (Schmidt's Jahrbiicher, 1867, 135, p. 81, and 140, p. loi). 
 
 32. — Robin. Des Vegdtaux qui croissent sur les animaux vivants, 
 Paris, 1847, p. 42. 
 
 33. Histoire naturelle des v^gdtaux parassites qui 
 
 croissent sur rhorame et sur les animaux vivants, Paris, 1853. 
 
 34. — Sternberg American Journal of Medic. Science, 1884 — 5. 
 
 35- — Vignal. Recherches sur les microorganismes de la bouche 
 (Arch, de physiol. norm, et pathoL, 1886, n. 8). 
 
 36. — Zopf. Zur Morphologic der Spaltpflanzen, Leipzig, 1882. 
 
 37- Die Spaltpilze, Breslau, 1883, p. 80, etc. 
 
 38.— Zurn. Die Schmarotzer, etc., Part II., Weimar, 1887—89. 
 For the works of David and Billet, see note at the end of 3rd paragraph. 
 
Plate 3, 
 
 
 
 
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AND CONTENTS OF THE MOUTH. 146 
 
 ®n Xeptotbriy IRacemoea. 
 
 By Filandro Vicentini, M.D., Chieti, Italy. 
 
 THIRD MEMOIR 
 
 ©n tbe Crsptogamfc jflora of tbe ^outb. 
 
 Translated by Professor E. Saieghi. 
 
 Summary. 
 § I. — Introductory and Bibliographical Notices. Eeference 
 to the previous Memoir. The Works of Pommay, 
 David and Billet. 
 
 § 2. — New observations on the fructification of the normal 
 parasite of the mouth {Leptothrix racemosa ?). Various 
 elements and aspects of the grape bunches. Fructifi- 
 cation, by temporary spores {sporids}), and by persistent 
 spores (teleutospores ?). Varied appearances and forms. 
 Conclusion. 
 
 w 
 
 §1- 
 
 INTRODUCTORY AND BIBLIOGRAPHICAL NOTICES. 
 REFERENCE TO THE PREVIOUS MEMOIR. 
 
 -"E have often lamented, in many works on Bacteriology, 
 the deficiency of ascertained opinions upon the Natural 
 History of micro-organisms. Without an adequate know- 
 ledge of the evolutionary cycle of bacteria in nature, from the 
 origin to the varied phases and their complete morphological 
 evolution, we could not even know whether each single bacterium 
 or bacillus, found in a determined material, is an isolated entity, 
 a real and proper living individual, or whether the varied Bacteri- 
 ological forms are but membra disjecta, dissevered and scattered 
 
146 BACTERIA IN THE SPUTA 
 
 particles, cellules, a series of cellules, or organs of fungi or superior 
 alga. 
 
 We entertained this doubt, in regard to the microbes of the 
 mouth and of the sputa, not through preconceived ideas, but after 
 a series of researches ; and we were led to suppose that the com- 
 mon progenitor of all, or nearly all, the bacterial forms of the 
 mucous cavities was probably an alga or fungus which thrives in 
 those parts, the Leptothrix buccalis of Eobin. 
 
 But this first point of the Microphytology, namely, the natural 
 phases of micro-organisms from the scattered or rudimentary forms 
 to those of their complete development, is exactly the part mostly 
 overlooked by bacteriologists, and especially by pathologists ; 
 whilst, in our opinion, it is the most important, and, as it were, the 
 basis of all the applications to the pathogenesis of the infectious 
 diseases (which is not always the same as that of the experimental 
 infections), and without which there would lack the necessary 
 foundation ; as the endeavours to investigate the contaminating 
 properties of different isolated bacterial forms, through the inocu- 
 lations, without ascertaining first, or at least in part, what are, in 
 reality, the bacteria in themselves, and which, in their natural phases 
 of life, might be the cause of premature deductions, which a more 
 accurate morphological research would afterwards upset. 
 
 Referring to the two Memoirs on the Sputa of Whooping- 
 cough and on the Bacteria of the Mouth and Sputa, we shall 
 simply repeat here our conclusions on the following points : — • 
 
 " (a) That the bacteria of the sputa, reputed pathogenic (e.g., the 
 Pneumococcus and Bacillus of Koch), probably are nothing but so 
 many disseminations, more or less modified, of the same microbes 
 and parasitic elements which normally thrive by hundreds of tril- 
 lions in the buccal and nasal cavities. 
 
 {V) That probably the contaminating properties of bacteria in 
 general (such as result from the inoculating experiments) are not 
 to be held as original, but dependent upon simple modifications, 
 occasioned, in single cases, by the nutrient substratum, more or 
 less alterable, on which they fall ; or upon the peculiar conditions 
 that their germs meet with in morbid points. 
 
 (f) That although to a certain extent the usual experiments of 
 culture and inoculation may be able to determine the pathologica - 
 
AND CONTENTS OF THE MOUTH. 147 
 
 entity of any forms of bacteria, they are yet quite unable to deter- 
 mine the botanical entity ; in other words, they cannot individualise 
 and qualify their species, as it is possible, generally speaking, to 
 get from the simple disintegration of the same complex micro- 
 organism, different inferior forms or living particles, scattered, 
 eventually endowed with varied contaminating properties,, and apt,, 
 mostly, to reproduce the original micro-organism ; but that, even 
 remaining sterile or simply multiplying in rudimentary condition, 
 does not constitute true distinct species. 
 
 (d) That in particular, the bacteria and bacilli of sputa do not 
 constitute complete micro-organisms or true and proper living indi- 
 viduals ; but they are simply severed and scattered particles, or 
 inferior forms of only one microphyte. 
 
 (e) That this microphyte lives normally, not simply in the nose 
 and mouth, but even in the external genitals, and, in given con- 
 ditions, attains its higher phases of development (analogous to 
 those of fungi or dioecious &\g<&), whose adult forms we have delin- 
 eated in the preceding plate. 
 
 To carry on these investigations, we have made use of a 
 homogeneous immersion objective of i/i 8th inch {v^yava. v^2 
 N.A.), by means of which we have been able to detect, in the 
 parasite in question, the existence of those superior phases of life 
 undescribed and unforeseen by other investigators. In fact, we 
 imagined that some productions in the shape of tufts, were male 
 inflorescences ; whilst, in more numerous productions, we thought 
 we recognised true fructifications or sporifications of this parasite 
 (either fungus or alga) in the shape of so many cumuli of small 
 spores taking a brilliant colour with aniline, placed in hne at the 
 exterior of the fertile filaments, and disposed in six longitudinal 
 series. This fructification by spores must be distinguished from 
 the gemmules of reserve or production [of bacteria enclosed in the 
 filaments. Our little spores seemed to be adhering to the 
 stalk through an amorphous, viscid substance, or glair, which 
 hardly becomes colourable. 
 
 Although with that objective we could not succeed in seizing the 
 further details of its structure, since then we supposed there were 
 not wanting some real peduncles (sterigmata or engrafting threads) 
 of the single sporules on the central stalk, so slender and trans- 
 
148 BACTERIA IN THE SPUTA 
 
 parent that they could not be detected with the optical instruments 
 employed at that time. Therefore, having left the matter unde- 
 cided, we proposed the name of ears for those fructifications. 
 
 The preceding Memoir was in the press when we kindly re- 
 ceived from Messrs. Bezu, Hausser, and Co., a new 1/2 sth inch 
 homogeneous immersion objective ; this objective revealed at 
 once the existence of the predicted peduncles or engrafting threads, 
 so that we were able to give a previous hint of it at the time, 
 proposing then to change the first name of ears into that of bunches 
 (as such are really the fructifications of Leptothrix buccalis). We 
 then promised to deal more specially with the subject in a new 
 communication. 
 
 We will, therefore, relate the most saUent facts connected with 
 the fructifications and other superior phases of our parasite, and 
 delineate the more important types of it, making use only of two 
 stains, gentian violet and an acidulated solution of iodine, discard- 
 ing all other stains. It would be inopportune now to go more 
 deeply into this morphological study, without having first con- 
 firmed the essential points which more exactly concern those adult 
 phases investigated and described by us for the first time. 
 
 The objective adopted for these new researches, and of 
 which we gave a hint in the note above referred to, is not 
 included in the last catalogue (1891) of its constructors. It is 
 a very solid objective of N.A. i'3o, the immersion fluid being 
 vaseline oil. It has an equivalent focal distance of i mm., pro- 
 vided with a correction collar, and possessing a proper or initial 
 magnifying power of 180 diameters, which, with the various 
 Huyghenian eye-pieces, magnifies from 820 to 3,100 diameters. 
 The working distance is relatively considerable. The images are , 
 perfect over the whole field, even with the No. 6 ocular, so that 
 with it may be detected from forty to fifty red corpuscles of the 
 blood (each one of them about 26 or 27 m.m. in diameter) in the 
 same visual field and plane ; but, beside its great magnifying 
 power, it is to be commended for its resolving power, which is not 
 inferior to that of apochromatics. The instrument we have made 
 use of is the No.VII. inclined stand of the same makers, with a 
 stage one decimetre square, revolving with the superior part 
 round the optical axis. The stand has an Abbe condenser of 
 
AND CONTENTS OF THE MOUTH. 149 
 
 1-40 N.A. (with centring and focussing motion), and is provided 
 with an iris diaphragm and dark-ground illumination. 
 
 Before proceeding to demonstrate our new observations, it is 
 necessary to complete the Bibliographical notices briefly described 
 in the previous Memoir. We made a point of perusing all the 
 publications upon this subject that we could obtain, in order to 
 ascertain whether others had anticipated us, or met with analogous 
 results, in regard to the superior phases of Leptethrix. We will, 
 however, only touch upon three works, published from 1890 to 
 1891 ; the more so as the same works (although silent on the 
 above-mentioned phases) offer us, on the other hand, points of 
 agreement with our various observations and views on the inferior 
 phases of microbes in general, and in particular of those of the. 
 mouth and sputa. 
 
 Article by Pommay. * 
 
 One of the most important points of the modern bacterio- 
 logical doctrines is to know whether, in the experiments of the 
 laboratory, the virulence of certain materials or cultures, contain- 
 ing bacteria, is to be held as original and part of the microbes in 
 question, or simply an accidental acquisition. Another point, not 
 less important, is also to know whether, in certain cases, the 
 virulence as such is not to be assigned to the material in which 
 they thrive, or to their products of secretion, rather than to the 
 microbes themselves. 
 
 The present work by Pommay is not the result of original 
 researches, but rather a retrospective review of many previous 
 observations gathered from different authors. In the first part the 
 author largely deals with certain conditions, which, although 
 extraneous to bacteria, are nevertheless to be held as the true 
 agents of the virulence. It has been proved that the inoculation 
 of substances secreted by the bacteria often gives rise to nearly 
 the same morbific phenomena attributed to the relative microbe 
 as such. Pasteur, by inoculating the sterilised liquid cultures of 
 chicken cholera, has reproduced the principal symptoms of that 
 disease. As much can be said of the Bacillus pyocyaneus and the 
 
 * H. Pommay, De Porigine et des conditions de la mrulence dans les 
 maladies infeciieuses (Annales de Micrographie, III., pp. 220—74.; 
 
150 BACTERIA IN THE SPUTA 
 
 cholera bacillus (Gamaleia) of the staphylococcus (Christmas), 
 and the Bacillus anthracis (Hankin), etc. To these examples more 
 recent ones might be added, but it is enough to point out the case 
 of the tetanus-virus. After the experiments of Gaillard and 
 Vincent, Kitasato in a recent article affirms that the cultures of 
 tetanus bacilli, filtered after Chamberland's Method, produce 
 absolutely the same tetanizing action as the cultures still containing 
 bacilli.* 
 
 " Virulence," concludes Pommay, " is not at all a necessary 
 function of microbes ; the secretion is an essential act of life of the 
 cellule .... but the nature of the secretion depends only upon the 
 total of the specific conditions of each micro-organism, and follows 
 up its variations in such a manner as to be more or less influenced 
 by the various changes of the external agents " (pp. 221 — 22). 
 
 At this point Pommay touches upon some particulars concern- 
 ing the influence of the external agents upon the virulence, the 
 dissolving power and other properties of bacteria, which we do not 
 consider. "Virulence," he adds, "namely, the secretions of poison- 
 ous matters, may be increased, reduced, and even suppressed by 
 natural agents, or by artificial means. We can, therefore, affirm, 
 that virulence is nothing but a contingent function of the micro- 
 organisms, that it depends chiefly upon the external agents, and the 
 microbes do not evolve from themselves or their own substance, 
 but from substances on which they thrive and multiply. The soils 
 or media of cultures may, in respect to this, be divided into indif- 
 ferent, positive, and negative soils. The indifferent soils have no 
 action whatever upon the functions of bacteria; if these are 
 virulent they secrete their virus, and if not virulent, they do not 
 become so. The positive soils do not only supply the materials 
 of virulence, but they modify the microbes so that the virulence, 
 occasionally lost by them in the negative soils, is revived. In the 
 negative soils the virulent microbes lose the aptitude of secreting 
 poisonous matters, and it is only through passing into a positive 
 soil, that they will become virulent again. 
 
 * Zeitschrift far Hygiene^ X. , page 267. We, however, begin to suspect 
 that an eventual passage of very minute bacterial particles through the porous 
 porcelain takes place, V. Freudenreich, De la fermMiliti des filtres Cham- 
 berland d,VSgard des bactiries. {Annates de Micrographie, Vf . , p. 5S9) F.V. 
 
AND CONTENTS OF THE MOUTH. 151 
 
 In conclusion, the virulence may, in a certain manner, virtually 
 exist apart from the microbes; these embody it, and set it to work ; 
 tt is an acquired and transitory function of the pathogenic micro- 
 organisms, and not an essential function inherent to their existence, 
 and it is subject to different external influences" (pp. 227 — 28). 
 
 The author, afterwards, dwells upon the changes undergone by 
 bacteria in the air, water, and soil. He infers that the pathogenic 
 bacteria are found in a very small number round about man, but 
 that it is necessary to look for them in man himself. He here 
 gives instances of pathogenic bacteria found in the mouth, nose, 
 and in normal sputa, in the same way already described by us 
 in our memoirs. 
 
 In the second part, Pommay begins by observing that simple 
 saprophytes may evolve themselves in the sense of virulence, 
 adapting themselves to new conditions of existence, and that 
 infectious diseases, whilst partly proceeding from pathogenic germs 
 evolved in individuals already invaded, can, on the other hand, 
 proceed from germs originally inoffensive, having become virulent 
 outside the economy, through the influence of the nutrient substra- 
 tum. The adaptability of micro-organisms is so unlimited, and 
 the time required for their metamorphoses so short, that the passage 
 from one phase to another seems to us instantaneous; consequently 
 we incline to make two different individuals of forms which, in 
 reality, proceed the one from the other. 
 
 It is quite probable that the bacteria considered pathogenic 
 may not be, and have not been always, such, but may have lived 
 previously, and may still live in process of time, as simple sapro- 
 phytes. The author thinks that the latest researches strengthen 
 that probability to a degree of certainty. 
 
 The bacillus of Lustgarten, considered to be the bacillus of 
 syphilis, is so like the bacillus of smegma as to be held for the 
 same normal bacillus, modified in its action, after having long lodged 
 in the materials of secretion heaped up and fermented upon the 
 genitals. A normal bacterium of the urethra is probably the 
 progenitor of the Gonococcus of Neisser (see our preceding notes 
 and memoirs). The bacillus of diphtheria exists in the condition 
 of a simple saprophyte in the mouths of healthy children (Eoux 
 and Yersin). Likewise the bacillus of Eberth, the specific germ 
 
152 BACTERIA. IN THE SPUTA 
 
 of iko-typhus, is considered to originate from Bacterium coli com- 
 mune (Rodet and Roux, of Lyons). 
 
 " The bacillus of Lustgarten, the Gonococcus of Neisser, the 
 bacillus of diphtheria, and the bacillus of Eberth, would, conse- 
 quently, be found (concludes the author) in us and upon us, in the 
 state of saprophytes, thriving inoffensively upon those same 
 parts that they attack at the moment of the virulence. From 
 such similarity of seats, we think of the relation existing between 
 the saprophyte and pathogenic microbes, or rather of their trans- 
 formation sur place " (p. 262). 
 
 This idea is strengthened by many historical arguments. 
 
 Many infectious diseases which are of more frequent occurrence 
 nowadays, did not formerly exist ; some, which were once com- 
 mon, have now disappeared, or tend to do so ; others show alter- 
 natives of activity and repose. A secluded life, for instance, has 
 favoured, in our times, the appearance of typhoidal diseases and 
 the spreading of tuberculosis. 
 
 Yellow fever is comparatively recent, having appeared since 
 the shores of the Gulf of Mexico and the Great Antilles have 
 become covered with populous centres ; and, unless we presume 
 a spontaneous generation of new species of micro-organisms, we 
 are bound to admit that the specific microbes of yellow fever (if 
 to this disorder a parasitical origin is to be assigned) are only a 
 transformation of other common pre-existing microbes, consequent 
 upon the contamination of the soil, the overcrowding population 
 with the co-efficients of climate, misery, etc. 
 
 Leprosy (formerly so common) is now confined, in European 
 countries, to very few localities, and even where it appears does not 
 make many victims, having nearly lost its virulence. We must, 
 therefore, admit an attenuation, and also presume a successive 
 disappearance of the virulent action of its bacillus, etc. 
 
 Other infectious diseases alternately appeared and disappeared. 
 A singular instance is found in the sweating disease which appeared 
 for the first time in i486, and four times up to 1530, when it 
 finally disappeared. We cannot imagine that the germs of the 
 first epidemic had been preserved until the appearance of the 
 second, and so forth ; but we must argue that the germs were always 
 present, at every epoch, only becoming virulent at intervals. 
 
AND CONTENTS OF THE MOUTH. 153 
 
 Analogous cases are exhibited by a similar disorder in Picardy. 
 Exanthematous typhus broke out in Algiers in 1867, without 
 being imported. 
 
 We must, therefore, according to Pommay, distinguish the 
 period of formation of an infectious disease from its period of 
 diffusion. In the first period we have the evolution of common 
 bacteria in the sense of virulence ; in the second, the multiplica- 
 tion of the germs themselves, owing firstly to their transmission 
 from one to the other affected organism, and secondly to their 
 extraordinary multiplication in excrements and secretions, or in 
 infected corpses. 
 
 Setting aside the considerations concerning hygiene, into which 
 the author then enters, we will simply give his final conclusions, 
 which so nearly agree with our own views : — 
 
 I. — " The virulence is not a primitive, original, and necessary 
 condition of pathogenic microbes ; it is an acquired, contingent 
 condition, which proceeds from the evolution of certain microbes, 
 in certain conditions and soils. 
 
 II.—" Of Culture media, some- increase or create the virulence; 
 others attenuate or extinguish it ; others are indifferent — that is, 
 neither modify nor produce it. 
 
 III. — " The pathogenic germs proceed either from morbid 
 points in which the virulence is acquired or maintained, or from 
 individuals affected with the disorder due to such germs. The 
 prophylaxis, to be called certain and complete, must, on one side, 
 ascertain and suppress the conditions producing the virulence ; 
 and, on the other, suppress or destroy the germs proceeding from 
 the diseased bodies, or impede the propagation of them. 
 
 IV. — " Water being only an intermediate between the organism 
 and the pathogenic germs, the prophylaxis based upon the purifi- 
 cation of water is uncertain and incomplete. 
 
 V.—" The pathogenic germs, taking up their virulence in the 
 soil or in the organism, must be attacked in those two fields ; the 
 purification of the waters coming next. 
 
154 bacteria in the sputa 
 
 The Work of David.* 
 David, late director of the Odontological School of Paris, is 
 already known by other works. The one to which we now refer, 
 may be properly called a work of compilation, following in the . 
 footsteps of Miller, and, although very valuable of its kind, helps 
 in a measure to complete Miller's work. 
 
 In the first chapter he gives a historical summary upon the 
 microbes of the mouth from Leeuwenhoek to the present time, 
 in which he especially mentions the last works of Pasteur, 
 Netter, Vignal, Miller, Blake, Galippe, and Biondi, quoted 
 by Miller in various places. The observations of Netter and 
 Vignal upon the presence of pathogenic microbes in the mouth 
 of healthy subjects are particularly worthy of notice. Vignal 
 found in his own mouth two staphylococci, and such re- 
 searches agree with those of Sternberg on the presence of the 
 pneumococcus and the tubercular bacillus of Koch in healthy 
 subjects (Comp. with our work on the Sputa of Pertussis). 
 
 Omitting the brief notices of Microphytology and Biological 
 Chemistry, hardly touched upon by the author, we come to the 
 second chapter (description of the common microbes), where are, 
 in the first place, exhibited the forms of Bacillus subtilis, Bacterium 
 termo. Bacillus amylobacter (or butyric ferment), the bacillus of 
 potatoes and of the Vibrio rugula. 
 
 In the number of common bacteria, the author even includes 
 Spirilla and Spirochete, although he himself, further on, speaks of 
 their infectious action upon the tissues and circumambient glands, 
 already recognised by Verneuil and by Clado. 
 
 We have elsewhere treated of the genesis of Spirilla 
 and Spirochcete, though omitting an important question of 
 their biology, which we now find hinted by David. That question 
 concerns the possible affinity of the Spirochcete with the brilliant 
 granules, their staining with aniline, and vibrating with a dancing 
 motion, which are lodged in swarms within the salivary corpuscles 
 (see Fig i, h). Rud and Arndt suppose that such dancing gran- 
 ules are nothing but germs of Spirochcete, temporarily sheltered 
 
 ^ Th. David, Les Microbes de la bouche, prkedS d'une lettre preface de 
 M. L. Pasteur, Paris, 1890, 8vo, pp. xvi.— 302, with 113 figures intercalated, 
 in colours and in black. 
 
AND CONTENTS OF THE MOUTH. 155 
 
 in the salivary corpuscles, from whence, being afterwards set 
 free, they would constitute germinating spores for producing new 
 Spirochoete. But the affinity of those dancing granules with the 
 so-called microzymes, and with bacteria in general, had been al- 
 ready foreseen by other observers, as was remarked by Beale.* 
 At present that affinity would be also connected with the fact, 
 not less important, of the normal nesting of bacteria within the 
 white corpuscles of the blood, ascertained by Baumgarten, which 
 is diametrically opposed to the hypothesis of phagocytes.f 
 
 David proceeds to describe, the Bacillus tremulus of Rappin, 
 the characters of which we have studied elsewhere; in our opinion, 
 it is one of the male elements of Leptothrix buccalis. 
 
 We are surprised that David admits the presence of Lepto- 
 thrix only in the Carnivora; whilst a nearly analogous parasite 
 has been recognised in most domestic herbivora and in swine 
 by Miller and other investigators. But the author is right in 
 doubting the separate entity of B. buccalis maximus and Lepto- 
 thrix buccalis maxima of Miller ; and this wholly agrees with our 
 views. 
 
 With regard to the pointed granules contained within the fila- 
 ments of Leptothrix, the author entertains the same opinion as 
 Robin, namely, that they might be spores. We, on the contrary, 
 incline to hold them as gemmules of reserve, destined, in the sev- 
 ered or truncated filaments, to become ellipsoidal bacteria, or even 
 dumb-bell bacteria, according to their respective position in the 
 envelope ; whilst the real spores should be found on the top and 
 outside of the fertile filaments. 
 
 The observation of Rappin (p. 67 of this work) is rather impor- 
 tant respecting certain filaments of Leptothrix found near their 
 engrafting point, in continuity with some little chains of bacteria. 
 David only sees in it an optical effect, due to the superposition of 
 rosaries of Streptococci with slender filaments of Leptothrix ; but, 
 in our last memoir, we have already described and delineated an 
 identical fact (preceding memoir. Fig. 9, d), the same unquestion- 
 
 * Beale, The Microscope in Medicine^ London, 1878, p. 272. 
 
 t Baumgarten, Zur Kritik der Metschnikoff'schen Phagocytentheorie 
 (Zeitschr.f. Klin. Med. Bd. XV., 1888.^ Ueber das Experimentum Cntcis 
 der Phagocytenlehre ^Zeigler's Beitrage 0. path. Anat. u. allg. Path. Bd. 
 VII., 1890, p. \.) 
 
156 BACTERIA IN THE SPUTA 
 
 able continuity of filament with the small chain, which will lead 
 us to the genetic identity of the two forms ; identity at all events 
 in harmony with the other forms or morphological phases of 
 Leptothrix. 
 
 David subsequently deals with the fungi of Biondi and of 
 Vignal already mentioned in our last memoir. At the close of the 
 chapter he hints at the digestive action of the buccal microbes, 
 and then at their fermentative action upon lactic acid, mannite, 
 dextrine, butyric acid, and other substances (see quoted Memoir). 
 
 The third chapter {Pathogenic microbes found in saliva) deals, in 
 the first place, with the forms of the pneumococci of Fraenkel and 
 Friedlaender clearly detected in the mouth even of healthy subjects. 
 On this point we refer to our two previous memoirs, and wish only 
 to state that the casualty or accidentality of the pneumococcus 
 of Friedlaender was recently demonstrated by Gamaleia*, and 
 we may, some day, obtain the same results with the pneumococcus 
 of Fraenkel. Probably such diplococci, incapsulated or bare, are 
 but a condition or particular phase of certain particles or articula- 
 tions of the same normal parasite of the mouth; a common 
 condition of other bacterial species, which Billet calls the zobglxic 
 state, as we shall see later on. 
 
 This chapter ends with the description of the so-called microbes 
 of suppuration {Streptococcus pyogenus, Staphylococcus pyogenus, 
 white and golden). 
 
 In the fourth chapter (Pathogenic microbes found in buccal 
 and dental affections), the author deals, firstly, with the tubercular 
 bacillus. That the bacillus of Koch was also liable to decolouri- 
 sation with nitric acid (at J) and alcohol, although in a degree 
 (generally speaking) less than other bacteria and bacilli, it was an 
 acknowledged fact even by Jacksch and Hunter Mackenzie, as we 
 have said before. The opinion of David is virtually nearer to 
 that of the authors mentioned. In the first place, the solution of 
 nitric acid, recommended by him in following the method of Ehr- 
 lich (p. 135) for decolovirising the preparations made by dry 
 processes, is found notably weakened ; it is no longer a J solution, 
 but rather a J or J. Secondly, the immersion in the nitric bath 
 
 * Gamaleia, Annates de VInstitut Pasteur, 1888. 
 
AND CONTENTS OF THE MOUTH. 157 
 
 has been gradually shortened. For Jacksch it was only a question 
 of reducing the violet to green ; for Hunter Mackenzie the length 
 of thirty seconds could not be exceeded ; and now David does 
 not allow the nitric bath (although doubly diluted) to last more 
 than two or three seconds. Dr. Kauffman, of Cairo, has lately 
 found that the bacilli of Koch, coloured with carbolic fuchsine, 
 retain the tint only five minutes, if washed in hot or boiling water. 
 He uses it instead of nitric acid and alcohol, but only for two 
 minutes.* 
 
 As we see, the opinion that the bacillus of Koch had a very 
 special retentiveness for aniline has been gradually losing ground. 
 At any rate, this is not the only bacillus which fixes tenaciously 
 the aniline, and we exhibited specimens, especially of diplococci, 
 in long envelopes. The bacillus of smegma and of leprosy have 
 the same property. Gottstein and Bienstock thought that the re- 
 sistance to decolourisation in those bacilli might depend upon a 
 protecting envelope of a fatty substance ; and in fact, Grigorjew 
 cultivating other bacilli {Bacillus anthracis, B. subttlis, Clostridi- 
 um butyricum. Bacterium termo, and the two Staphylococci), in 
 fatty media, affirms to having found them equally resisting. t 
 
 Of the other microbes grouped by the author under this same 
 heading, we shall only mention the Micrococcus tetragenous, Oidium 
 albicans, Actinomyces, and then the fungi of Miller (including the 
 Jodococcus vaginatus and the Spirillum sputigenum), and the already 
 mentioned fungi of Biondi. At page 206, a Leptothrix pusilla is 
 dealt with, described by Klebs as the ceiling of the dental tartar, 
 but it is discarded by the David. 
 
 The fifth and last chapter (Practical and therapeutic deduc- 
 tions) does not concern bacteriology. The appendix on The 
 Microbes of Influenza, preceding the last studies of Pfeiffer, arrives 
 at unsatisfactory results. 
 
 * Modern Medicine and Bacteriological World, May, 1893, p. 126. 
 
 t Gottstein, Deutsch med. Wochenschr., 1886. Bienstock, Fortschr. 
 d. Med., IV., 1886. Grigorjew, Ruskaja Medizina, 1886. 
 
158 bacteria in the sputa 
 
 The Work of Billet.* 
 
 Billett, Surgeon-major in the French army, had already pub- 
 lished other works' upon the Sporulation of Cladothrix dichotoma, 
 upon Bacterium urea (1885), upon B. laminaria and B. Balbiani 
 (1888). The present work, for which the Institute of France 
 awarded the Montaigne prize for 1890 upon the report of Bornet, 
 has been published in full in the Bulletin Scientifiqiu de la France 
 et de la Belgique, by Prof. Giard. It is divided into an introduction 
 and four sections (one on each of the described species), besides 
 a rich bibliographical Appendix, comprising 662 works, mono- 
 graphs, or groups of articles, upon that or kindred subjects. 
 
 Examining the simple phases, which we call inferior, of the 
 micro-organisms, Billet goes to demonstrate a thesis analogous to 
 ours on the Leptothrix buccalis, and to establish a similar genetic 
 identity amongst the varied bacterial and bacillary forms proceed- 
 ing from four other species of micro-organisms (Cladothrix dicho- 
 toma, Bacterium Balbiani, B. osteophilum, and Leptothrix parasitica, 
 Kiitzing, a parasite of the superior sea-weeds), and he comes to the 
 conclusion that such micro-organisms may even be algse analagous 
 to CyanophycecB, and that, like those, they present themselves 
 either under the form of a fibrous sprout, or of isolated and motile 
 elements, as the hormogonia of the Nostocacece. The Cladothrix 
 dichotoma resembles in the sporulation (which we call gemmula- 
 tion) the Nostoc ellipsosporium, described by Bornet. The zooglceic 
 state reminds us, in its turn, of analogous forms, described as the 
 ChroococcacecE (a group of the same Cyanophycese). The absence 
 of the chlorophyll is not sufficient to relegate such micro-organisms 
 to the family of fungi or schizomycetes, after that Van Tieghem 
 and Engelmann have demonstrated the pigment of certain 
 Bacteriacese of a green and purple colour to be a chromophyll 
 (analogous to chlorophyll), by means of which such plants decom- 
 pose, on one side, the carbonic acid under the action of light, and 
 on the other, produce the synthesis of the hydrates of carbon. 
 
 In the introduction the author recapitulates the history of the 
 
 * A. Billet, Contribution d, Vitude de la Morphologic et du diveloppement 
 des Bactiriacees, Paris, 1 890, 8vo, pp. 288, with 19 figures and 9 plates, some 
 coloured. 
 
AND CONTENTS OP THE MOUTH. 159 
 
 Baderiacem : a group of organisms (he says) of which the study 
 has much interested biologists ; still, there are members of the 
 vegetable kingdom whose history is so scantily cleared up. Under 
 the generic name of BaderiacecB adopted by Van Tieghem, the 
 author designates the whole of the microbic forms commonly 
 called Schizomycetes or Schizophytes ; the morphological study of 
 which, inaugurated by the first investigators (from Leeuwenhoek to 
 Hoffmann) has been afterwards abruptly abandoned in order to 
 follow, almost exclusively, the phenomena connected with their 
 biological action on fermentations, putrefactions and infectious 
 diseases, as investigated by Pasteur and Koch. The small 
 number of morphologists who continued the morphological research, 
 soon divided into two fields : one maintaining with Cohn 
 the immobility of bacterial forms ; the other sustaining with Eay 
 Lankester, Cienkowski, and Zopf the theory of pleomorphism or 
 passage from one form to another, provided that this passage be 
 not taken, nowadays, as a transmutation from one species to another, 
 but as a series of various phases common to appurtenances of more 
 complex species. 
 
 The first investigators examining the bacterial forms in Uquid 
 substrata, became acquainted only with their isolated condition, 
 and they created the species of Cocci, Bacteria, Bacilli, Vih-ios, 
 Spirilla, and Spirochete. The first modification of these absolute 
 ideas is found in Eobin, who, in the Leptothrix buccalis, shows the 
 affinity between the Bacilli and the uncoloured filamentous algce, 
 purposely called Leptothrix by Kiitzing. And,- in fact, we have to 
 admit that even other species (viz., the genera Beggiatoa, Clado- 
 thrix, Streptothrix and Crenothrix) assume, besides the dissociated 
 forms, the filamentous forms. Cienkowski (1877) demonstrated 
 that the forms of bacteria were transformed into cocci, and that the 
 cocci in their turn assumed the form of Leptothrix; he called the 
 attention of the microphytologists to the zooglmic formations, men- 
 tioned by Cohn in 1853. He verified this zoogloeic phase in the 
 genera Cladothrix, Crenothrix, and Leptothrix together with the 
 preceding phases. In 188 1 Zopf extended the like views to the 
 genus Beggiatoa; he showed that the filamentous forms can be 
 dissociated in Bacilli and Bacteria or assume curved, vibrionic, 
 spirillic or spiroch^tic forms ; he explained the genetic connection 
 
160 BACTERIA IN THE SPUTA 
 
 and demonstrated that certa,in zooglxic formations held as bacterial 
 independent forms, constitute in reality but one proper phase of 
 the same filamentous species. 
 
 The views of these authors, generally adopted for some of the 
 highest types (Cladothrix, Crenothrix and Beggiatoa) were not 
 extended to most of the bacterial species. The three mentioned 
 species formed as a distinct group, whilst for the others we enter- 
 tained the primitive views of Cohn. But later on new exceptions 
 were introduced, which have now so far multiplied as to become 
 the rule. The author quotes the three species of Proteus of 
 Hauser, the Helicobacterium of Escherich, the Bacterium carotarum 
 of A. Koch, the Bacillus brassicm of Pommer, the B. cxruleus of 
 Smith, the B. pyocyaneus of Fliigge, the Spyrobacillus Cienkowskii, 
 etc. Only for the ancient genus Micrococcus an absolute separa- 
 tion is still maintained, but (adds the author) the Pneumococcus 
 itself shows to Friedlaender, Pipping and E. Klein, gradations to 
 the rectilineal forms. The same may be said of Micrococcus urea 
 and of Micrococcus prodigiosus. 
 
 Billet should have mentioned the last work by Miller (Die 
 Mikroorganismm der Mundhohle), where his return to the preced- 
 ing views appears evident, but it would appear from the 
 bibliographical index that it was not then known to him. 
 
 " The theory of the genetic relations of the different bacterial 
 forms" (continues Billet) "is not simply based upon exceptional 
 cases and speculative views. It has been, and is yet sustained 
 by workers whose methods cannot be suspected, nor their talent 
 for observation and interpretation questioned. To us the morpho- 
 logical question of the Bacteriacese is even wider. The point is 
 not only to know whether such and such species present a succes- 
 sion of forms proceeding one from the other, but, above all, to 
 know whether the Bacteriacese have an evolutionary cycle in mor- 
 phological characters particular and constant to each species, the 
 general development of which depends upon a law common to the 
 greatest part of them. In fact, from data in our possession, we can 
 easily perceive that most of them pass through various phases dur- 
 ing their evolution, according to their conditions of life. Thus, in 
 a first phase, the bacterial elements are found associated in a 
 filamentous graft, more or less long ; in a second phase, these ele- 
 
AND CONTENTS OF THE MOUTH. 161 
 
 ments become free and motile ; in a third, they group themselves 
 m gelatinous masses more or less considerable, and which, to some 
 of them, seem characteristic. Lastly, in a fourth and last phase, it 
 may happen that the preceding filaments intertwine one with 
 another in coiled masses at times very voluminous. We describe 
 these four phases more particularly as the filamentous, dissociated, 
 interlacing {enchevUrS), and zo'dgloeic states." — pp. 21-22. 
 
 We cannot follow the author in the special chapters devoted to 
 each of the four species studied by him. We shall only show some 
 points which may, in our opinion, clear up certain questions 
 connected with the microbes of the mouth and sputa. 
 
 And firstly, the dissociated and zoogteic states, considered as 
 simple phases of filamentous bacteria render it possible that diplo- 
 cocci or pneumococci may have been derived from the normal 
 leptothrtcal filaments themselves, as we have noticed elsewhere. 
 
 In fact, if we look at the relative figures of Billet, reproduced by 
 us, we may take them, at a first glance, for true pneumococci rather 
 than for simple disjunctions or simple zoogloeic groups of the algae 
 studied by the author. In Figure 1 7, a represents a fragment of 
 six articulations of Cladothrix dichotoma in disjunction, magnified 
 to 320 diameters, which, except in its greater size, does not differ 
 from the pneumococci. In b are noticed analogous articulations 
 magnified to 1,600 diameters, in a more advanced stage of disjunc- 
 tion. In c, d are seen two specimens of the Bacterium osteophilum 
 in its passage to the zoogloeic state, magnified to 600 diameters. 
 In e is drawn a fragment of four articulations of the same 
 Bacterium in that particular zoogloeic form called by Billet the 
 Scorpioidal state. In/ is reproduced a small group of the zoogloeic 
 state of Leptothrix parasitica Kiitzing, magnified to 745 diameters, 
 and their perfect resemblance to the sheathed diplococci, reputed 
 specific, is not less evident. Other similar specimens are repro- 
 duced in Figure 18, « (stained with solution of iodine and after- 
 wards with methyl violet or fuchsine, magnified to 600 diameters^, 
 where are detected rectilinear forms of Cladothrix dichotoma, analo- 
 gous to the sheathed diplococci drawn by us in the preceding 
 plates (Fig. 2, /, Fig. 4, g), in their stage of passing to the 
 zoogloeic state. In b fstained first with vesuvine, then with methyl 
 violet and solution of iodine) is seen the beginning of the zoogloeic 
 
162 BACTERIA OF THE SPUTA 
 
 State of Bacterium osteophilum, magnified to 600 diameters ; m c 
 thebeginningof the scorpioidal state ; and in da. larger representation - 
 of the first state (1,600 diameters). In e (stained the same as a) 
 is' seen a zoogloeic group of Leptothrix parasitica, magnified to 
 320 diameters. All those figures clearly show their analogy with 
 the pneumococci. 
 
 The other point we think worth considering is the formation 
 (in Cladothrix dichotoma) of the so-called zoogloea terminalis pre- 
 dicted by Cienkowski and demonstrated by Zopf, under the name 
 of zoogloea ramigera. This form is particularly produced in the 
 cultural liquids containing animal substances in putrefaction. A 
 specimen oi arborescent zoogloectoi Cladothrix is seen in Fig. 19 with 
 2t branches upon a single peduncle (coloured like Fig. 18, ii, 
 magnified only to 120 diameters). In a, b (Fig. 20, similarly 
 coloured, magnified to 600 diameters) are drawn the ends of two 
 clavaled branches, in order to show their contents, which are 
 formed of bacteria of various sizes and shapes ; these are better 
 observed in (Tof the same figure, magnified to 1,050. diameters. 
 
 Now such forms simply heaped up must be distinguished 
 from the fructifications by ears or clusters, already described 
 in Leptothrix buccalis, for their arborescent disposition (in the 
 zoogloea), for the irregularity of their outlines, their much larger 
 size, the mixture of heteromorphic bacteria in the same mass, and 
 finally for the want of the internal stalk which is very apparent in 
 the ears ; and that, apart from the last fact verified by us of the 
 peduncles or engrafting filaments of the little sporules of Lepto- 
 thrix on the central stalk. Of such external sporulation upon cer- 
 tain fertile filaments and of the production of tufts (probably 
 male organs) upon other filaments, not a word or even a hint is to 
 be found in either Miller, David, or Billet. 
 
 Being obliged to pass over the brilliant researches of the author 
 upon each of his four species, we will here give a partial summary 
 of his conclusions. 
 
 I. — The various states of the Bacteriacece correspond to 
 a special grouping of their elements in relation to the conditions of 
 the nutrient substratum (composition, temperature, pressure, aera- 
 tion, &c.) Therefore nothing is fixed in the succession of those 
 phases ; one state can rapidly evolve towards another, or fail, or 
 
AND CONTENTS OF THE MOUTH. 163 
 
 remain unchanged in the so-called pure culture, viz., in a medium 
 of known and unvariable composition under the same conditions 
 of temperature, pressure, and aeration. 
 
 II- — 'Yh^ filamentous state is pre-eminently the vegetative state. 
 Here the bacterial elements are disposed in small chains or articu- 
 lated filaments (by division of a primitive element in a single direc- 
 tion) sometimes enveloped in a real filamentous sheath, sometimes 
 simply joined by bands of intersticial substance or glair (of the same 
 nature as the sheath) secreted from the bacterial element itself, and 
 which, when it is more abundant, involves the groups of the 
 zooglaic state. 
 
 Ill- — The bacterial elements of the filaments occupy the in- 
 ternal part under the three rectilineal forms in Leptothrix, Bacillus 
 and Bacterium ; and in the three curved forms. Vibrio, Spirillum 
 and Spirochoete ; forms differing between themselves but all pro- 
 ceeding from the primitive rectilineal element. It is no question 
 of the pleomorphism of the old hypothesis nor of change from one 
 species to another, but simply of change of state. 
 
 We pass over what the author positively states about the form 
 in Micrococcus. He only found it once in Bacterium Balbiani and 
 he believes that it proceeds from the same elements in Bacterium, 
 reduced to a minimum of length through a change of the culture 
 medium. In fact, by restoring the medium to its previous condi- 
 tions the primitive form was resumed. In Cladothrix and in 
 Bacterium osteophilum, on the contrary, the roundish elements pro- 
 ceeded from a retraction of the protoplasm evolving itself towards 
 the spore. On our own part it is enough to observe that in 
 Leptothrix buccalis the roundish forms are very common, and (in 
 all probability) proceeding from true spores. The four species 
 studied by Billet (for reasons which we shall not investigate), 
 did not exhibit a true sporulation (on the exterior of the fila- 
 ments) which, we think, would explain the total or partial absence 
 of the micrococci. 
 
 Eeturning to the filamentous state, the condition more apt to 
 develop it properly is, according to the author, on one side the 
 fluidity of the nutrient substratum, and on the other the nearness 
 to the surface. It is essentially an aerobic state. Another char- 
 acter which distinguishes it is that it generally has non-motile 
 filaments. 
 
164 BACTERIA OF THE SPUTA 
 
 \ 
 
 IV.—" The dissociated state is characterised by setting free the 
 constituent elements of the filamentous state. These elements 
 continue to disintegrate, but instead of uniting into series or 
 chains they separate and live isolated. At times, however, 
 they remain united in couples or in small chains, but dissimilar 
 from the preceding elements, they are essentially motile. Conse- 
 quently in the dissevered state all those forms will be found which 
 have been already described in the filamentous state. Physio- 
 logically, the dissevered state has a very important office to fulfil, 
 which is chiefly a phase of dissemination. Thanks to the mobility 
 and the active segmentation of its elements, it can in a short time 
 spread over a very extensive culture medium. Thus, it so hap- 
 pens, that under these very forms of isolated, dissevered or motile 
 elements the Bacteriaceae are more frequently met with, and are 
 better known. 
 
 V. — " The interlacing state constitutes a third phase of the 
 developing cycle of the Bacteriacea. It shows itself under the 
 form of filaments interlacing into one another like so many threads 
 in a piece of knitting. Physiologically, this is a transitory state 
 between the dissevered and filamentous states, or between one of 
 those two states and the zoogloeic state. 
 
 VI. — " The zoogloeic state is the last phase of the evolutionary 
 cycle. The elements of the preceding states group themselves 
 according to certain dispositions which, at a first glance, do not 
 exhibit any apparent order, but in reality they follow up a definite 
 order, which in the Bacteriacecs we have considered varies for each 
 species. The chief phenomenon is the secretion round each 
 •element of a viscid substance or glair, morphologically and 
 physiologically identical with the sheath involving the filaments. 
 By the same fact of the increase of this gelatinous envelope, which 
 surrounds them like a capsule, the elements become motionless." 
 
 The zoogloeic state afterwards behaves differently through the 
 various stages, which we will briefly notice. In the first stage 
 (corresponding to the form already described) each element 
 is enveloped in a proper gelatinous capsule, and the morphological 
 aspect of the elements entirely corresponds to that which 
 characterises the genus Hyalococcus, created by Schroter for 
 the Pneumococcus of Friedlaender. In a second stage of the same 
 
AND CONTENTS OF THE MOUTH. 166 
 
 zooglceic state, we have the forms of Merismopedia or Tetrads, and 
 in a third state the form of the Sarcina. To this form we think 
 should be related the straight and curved diplococci of types f, /, / 
 (Fig. 2), from which the so-called Gonoccocci of Neisser would then 
 be derived. Sometimes we have also the forms of Ascococcus or 
 Zooglcea terminalis, of which we have made a mention before. 
 
 Physiologically, the author believes that the zooglceic state con- 
 stitutes for a large number of the Bacteriacex, z. protecting phase of 
 the relative elements against the external agents, or of preparation 
 for more favourable conditions in which to be able to abandon the 
 gelatinous envelope in order to thrive again in the filamentous or 
 dissevered state. As regards the formation of endogenous spores, 
 we could not, without some reserve, adopt the opinion of 
 Billet, having already elsewhere expressed the idea that such 
 bacterial forms enclosed in the filaments, are only (at least in our 
 parasite) a development of the gemmules of reserve. Such gem- 
 mules are not, on the other hand, to be taken for the granulations, 
 considered by him as products of reduction, due to a degenerative 
 phase of the respective articulations. 
 
 From these remarks the author concludes that in order to 
 determine the bacterial species, it is no longer sufficient to 
 describe their isolated forms in the single soils or media; but 
 it is necessary to investigate whether any forms remain always 
 constant, even in different media. Then many forms held for- 
 merly as distinct species, will simply appear as accidental of the 
 same varieties. 
 
 The author lastly asks what are the probable affinities of the 
 BacteriacecB with other vegetal groups, and, having discarded (as 
 we saw before) the analogy with fungi, he brings them nearer to 
 the cyanophyceous algae (Nostocacea) according to the original 
 opinion of Cohn and the ideas of Van Tieghem and Bornet. And 
 for the following reasons : — (1) Their filamentous development in 
 one single direction, like the trichomes of the filamentous Nosto- 
 chinece; (2) their motion, analogous to that of the Oscillariacece , 
 (3) their dissevered state, similar to the Hormogones; (4) their 
 zooglceic state, analogous to the tabular sprouts of Merismopedia 
 and GlcEOcapsa; (5) their endogenous germination of the spores 
 (in the Cladothrix), analogous to that of certain nostocs. 
 
166 BACTERIA OF THE SPUTA 
 
 " Finally " (he adds), " we cannot do better than repeat the 
 words of Dallinger: — The bacteria are without much question ^a 
 botanical group, and to be thoroughly understood must be studied as 
 such. In order to know them and to be able to catologue their 
 species, as we have done for the other vegetal families, we have, 
 then, to investigate closely their phases ; and when we shall be 
 better acquainted with the true being of their species, we shall 
 be able to study more accurately their true office, their functions, 
 and physiological properties." — (pp. 208-18.) 
 
 When, in October, 1890, we presented the preceding Memoir, 
 we were not acquainted with the above-mentioned works. Seeing 
 now that our views are, at least in part, shared by these and other 
 competent investigators, we hope the opposition to the'^course we 
 have followed in these studies will vanish or be minimised, and 
 that the present remarks (although incomplete) will induce others 
 to initiate further and more accurate researches. 
 
 We pass now to describe our last investigations upon the 
 fructifications by clusters and the other superior forms of the 
 normal parasite of the mouth. 
 
 § 11. 
 
 NEW RESEAECHES OBT THE FRUCTIFICATION OF THE 
 
 NORMAL PARASITE OF THE MOUTH 
 
 (LEPTOTHRIX RACEMOSA?) 
 
 I TRUST it will not appear too pretentious on my part if I 
 propose for such an isolated form — or, better to say, for the tiny 
 plant which thrives (and appears to germinate and fructify) — on 
 our teeth, taken as a whole, the name oi Leptothrix racemosa, instead 
 of that of Leptothrix buccalis ; and this solely on purpose to quaUfy 
 it better, if it be true that the cryptogamic species must take their 
 name and character from the fructification (if any) rather from the 
 inferior or rudimentary appearances of their single particles, inci- 
 dentally incomplete, scattered, or isolated. 
 
 In describing the various aspects of the fructification in ques- 
 tion, we shall endeavour to group them in such a way that, 
 
AND CONTENTS OF THE MOUTH. 167 
 
 according to our view, there may be a better connecting of the 
 varieties ; but, in doing so, we do not wish to prevent a more 
 matured judgment. 
 
 Various Elements and Aspects of the Bunches. 
 
 To understand the various aspects which the ears or bunches 
 of Leptothrix racemosa present, we must value the different con- 
 stitutive elements of those clusters, which, according to our 
 observations, we consider to be four, in their natural order of 
 development. 
 
 The principal element, from which the other three proceed and 
 upon which they are formed, is the fertile filament, generally 
 slender, pale, ''containing internal parietal gemmules, invisible in 
 the iodine solution (simple or acidulated), but distinct enough in 
 gentian violet. For this kind of stalks we refer to the preceding 
 memoir. Then is it not strange that these fertile filaments should 
 be so slender, whilst the severed or truncated (Bacillus buccalis 
 maximus and Leptothrix buccalis maxima of Miller) appear much 
 thicker and with a strong iodine reaction. We attributed this 
 thickness, or woody state, to the retrocession of the germinal 
 matter, somewhat analogous to the action of the saps in pruning- 
 plants. Our parasite, being a vegetable, or rather a tiny plant, 
 and having to be studied as such, according to Dallinger,* it is 
 natural that the form of its stem should resemble that of the cone. 
 The stalk being, in fact, the proper organ of vegetation, destined 
 to support the upper Organs, it is natural that the lower part 
 should be stronger and that it should get thinner towards the top, 
 which, finally, must bear the fruits. It is identical with the 
 process of other plants. 
 
 However, whilst this is the general rule; we shall see, as we 
 proceed, that certain fertile filaments become rather an exception, 
 thinning' themselves similarly on the top ; but afterwards, on 
 reaching the base of the future fructification, they thicken in the 
 shape of a cylindrical club, colouring brilliantly with aniline, but 
 pale like the rest in iodine solution. We shall speak later on of 
 the probable meaning of this swelling of the stalk, recalling only 
 
 * Dallinger, The Microscopical Organisms and their Relations to Disease 
 (Journal R.M.S., iS8j.) 
 
168 BACTERIA OF THE SPUTA 
 
 on this subject the other apical swellings, already mentioned and 
 delineated in the preceding Memoir (Fig. 9, b c). 
 
 To this first- element (the internal stem) follows in order of 
 -formation the second, of peduncles or ingrafting threads,, destined 
 to bear the spores, like the sierigmata of many fungi. Such 
 peduncles are very pale, invisible with less powerful objectives, but 
 very distinctly observable (under certain conditions) with the 
 i/25th power just mentioned. They are short, funnel-shaped, 
 with a point on the stalk and the opening towards the spore. In 
 the young ears, being yet deficient or scanty in the secretion of 
 the viscid substance, the peduncles are more visible ; whilst in 
 the older ones they remain more or less opaque. 
 
 The spores constitute the third element, and are^of globular 
 form ; pale in iodine solution, more or less coloured in the aniline. 
 They are smaller in the younger clusters, on which, for this very 
 reason, the peduncles or sterigmata are more discernible'; whilst, 
 the spores becoming afterwards larger and covered with a more 
 viscid substance, the peduncles remain hidden. Besides, the spores 
 are not equally thick in all the fructifications ; but where thejspores 
 are thinner, as in Fig. 23, a, there the details mentioned are more 
 visible. The little or non-visibility of the peduncles may result, 
 not only from the opacity produced by the abundance and density, 
 or from the heavy colouring of the viscid substance, but even, 
 where this is thin and transparent, from the identity of the index 
 of refraction of the two elements. 
 
 The fourth element is the viscid substance ox glair, which we 
 hold to be the last to form. It proceeds from a sort of oozing or 
 secretion of the stalk or of the spores themselves, for, in the 
 younger ears, with yet small spores, it "appears thinner and 
 indistinct. 
 
 And these are the fructifications which exhibit a more striking 
 resemblance to real clusters, either in the solution of iodine or 
 when they are very slightly afiected by gentian violet. On the 
 other hand, in the ears with an internal swelling of the stalk, the 
 size of the spores, as well as the density and colouring of the 
 viscid substance, reach the highest degree, as we shall see later on. 
 That the viscid substance may proceed from the filament is 
 exhibited by the cited examples, in' other species, by Billet, as well 
 
AND CONTENTS OF THE MOUTH. 169 
 
 as in our parasite, by those filaments of Leptothrix buccalis maxima 
 and of ^a«7/w(5«c^a/wwax«wMj, which, witlnhenewi/25thobjective, 
 appear thoroughly enveloped in a hyaline sheath. That it may 
 proceed from the spores is shown by the example of the incapsu- 
 lated diplococci. However, between the exudation of the old 
 filaments and that of the ears there is this difference : that the 
 first has greater affinity for the acidulated solution of iodine, in 
 which it is better discerned, and the second for the gentian violet. 
 
 Keeping in view, on one side, these various elements, and, on 
 the other, the degree and different nature of the colouring, we may 
 fully explain the varied aspects of the fructifications. 
 
 Concerning the gathering and the preparation of the patina 
 dentaria and its treatment with the aniline colours, or with the 
 solution of iodine, there is little to add to what has been said in 
 the previous Memoir. We gave there the precise rules, in order 
 to obtain from the patina dentaria the greatest possible number of 
 fructifications, taking it before a meal or early in the morning. 
 AVe found afterwards that the fructifications are more abundant on 
 teeth with a thin film of tartar, particularly in the superior eye- 
 teeth. About the disintegration and colouring of the clods we 
 refer to what has been said. For the acidulated solution of iodine, 
 we placed on the slide, first, a small drop of lactic acid, disin- 
 tegrating in it afterwards the patina, and lastly adding to it one or 
 two drops of solution of iodine. It generally takes a quarter of 
 an hour to get a proper colouring. 
 
 In the preceding Memoir we suggested that the superfluous 
 liquid should be allowed to trickle down from the sides, after 
 applying the cover-glass ; but afterwards we discovered that it was 
 better to let most of the liquid evaporate before applying the 
 cover-glass, so as to avoid the wave of liquid caused by the pres- 
 sure, which might remove many bacteria and bacilli and several 
 isolated filaments and clusters set free from the tiny islands that 
 often supply the most instructive specimens ; blotting paper would 
 also take up many of the specimens. To eliminate the air-bubbles 
 keep the cover-glass on edge (straight up) with the two first fingers 
 of the left hand, so as to form an acute angle with the slide, at 
 two centimetres from the preparation ; then with a straight needle, 
 held in the right hand to support the glass, and with a bent needle 
 
170 BACTERIA OF THE SPUTA 
 
 pushing it on the specimen, lower it down gently on to the edge of 
 the preparation, when the air-bubbles will be set free. 
 
 If we wish to institute a comparison between the two stains 
 adopted in this work, we may say that gentian violet colours 
 briskly the little spores, but at the same time attacks and obscures 
 the viscid substance, so that, wherever it fully invests, the peduncles 
 or sterigmata are either not seen at all, or may hardly be distin- 
 guished. This happens even by applying very little tint. Never- 
 theless isolated fructifications slightly affected by the tint may some- 
 times be found in the preparation, and it is just upon one of these 
 occasions that we first detected the peduncles in question. For 
 this reason, the whole image, either of the single fructifications, or 
 of their branching filaments, is better seen in the anilines (see in 
 tlje previous Memoir the Figures lo, 12, 16); the solution of iodine 
 would hardly give the same result. This solution has little or no 
 action upon viscid matter ; therefore, the fructifications assume 
 with it a granular aspect, and if the spores are not too thick it 
 shows the peduncles better ; also because the solution of iodine 
 acidulated with lactic acid attacks relatively better the peduncles 
 themselves, whilst the gentian violet invades them less than other 
 elements. Consequently it happens that the solution of iodine 
 better exhibits the complei aspect of the clusters or the truncated 
 sterigmata. (See Fig. 23, 1^. J It has, however, the disadvantage of 
 not satisfactorily allowing the use of powerful eye-pieces, thus 
 limiting the enlargements ; whilst in the fructifications stained 
 with gentian violet, the details of structure may be (under favour- 
 able conditions) detected even with a No. 6 eye-piece, as is shown 
 in Fig. 24, magnified to 3,100 diameters. 
 
 Now from the concourse of these various circumstances, either 
 relative to the age of the single fructifications, and to the more or 
 less thickening of peduncles and spores, or to the quality and 
 degree of the colouring, we are able to obtain images conspicuous 
 in the whole, but with peduncles only partially or not at all visible, ' 
 or sometimes less conspicuous in the whole, but with quite dis- 
 tinct peduncles. The necessary conditions to the clear vision of 
 the peduncles in question may be summed up in the following 
 series : — a, proper optical instruments ; b, clusters still , young ; c, 
 rather thin spores ; d, a weak gentian violet ; or e, solution of 
 
AND CONTENTS OF THE MOUTH. 171 
 
 iodine. We have already said that the best images are obtained 
 from isolated fructifications fallen from clods in a clear field, and 
 It IS our intention in this work to consider the two colourings above 
 mentioned, apart from the use of other tints. 
 
 The fructifications in question can be observed by axial illu- 
 mination or by oblique hght. The best images of the clustered 
 forms and of the single sterigmata are obtained by axial illumina- 
 tion by properly adjusting the correction collar, and by centering 
 the iris diaphragm and the Ahh6 condenser along the optical axis. 
 On the other hand, the general relief of the ears and the position 
 of the spores in six longitudinal series, are better detected by 
 oblique light, by pushing aside the diaphragm and letting it after- 
 wards go round the optical axis, or substituting for it the dark 
 diaphragm, pushed on one side, and making the stage rotate with 
 the upper part of the instrument round the same axis. In the 
 latter case, a better effect is obtained when the ear is horizontally 
 disposed and is struck from behind by the pencil of light, almost 
 parallel with the stalk, beginning from the root, so that the - 
 luminous rays run along its axis. Then, focussing, the general 
 effect of the six series of spores becomes striking, the whole ear 
 takes a beautiful mulberry appearance, of which it is impossible 
 to give a satisfactory representation. 
 
 The figures which we have drawn are, however, sufficient to 
 give a proper idea of the peduncles in question. In Fig. 23, a, we 
 have represented a short and young ear, as seen stained with the 
 acidulated solution of iodine, magnified to 1,700 diameters; in it 
 the spores are thin, and its clustered form is most striking, as we 
 can even perceive posterior rows of little spores. In c (same 
 figure) is seen an older ear, thicker and longer (same staining, 
 and magnified 1,170 diameters) in which, however, the peduncles 
 are sufficiently distinct. In d, then (same staining and magnifica- 
 tion), is drawn a short fragment of a fertile stalk, found by chance 
 in one of the numerous preparations ; only two spores are seen at 
 its base, and higher up the peduncles still thick, but without 
 spores ; the rest of the ear is wanting.* In the above Fig. 24 
 (weakly stained with gentian violet, magnified to 3,100 diameters) 
 
 *The attenuated appearance of this stalk probably depended upon a flowing- 
 out of its sap or germinal matter owing to rupture. 
 
172 BACTERIA OF THE SPUTA 
 
 the spores are intact, their funnel-shaped peduncles are visible 
 through the viscid substance, moderately stained. The stalk 
 exhibits several gemmules of reserve. 
 
 The facts hitherto given (in support of the arguments 
 expounded or simply suggested in the preceding Memoir), which 
 everybody can verify for himself, are, in our opinion, sufficiently 
 conclusive to warrant us in affirming the existence of a really 
 external sporification or fructification of the normal parasite of the 
 mouth upon six longitudinal series of peduncles and spores. But 
 for a more evident proof, take the specimen drawn in Fig. 25. 
 This specimen, obtained by pure chance, amongst the number- 
 less preparations examined, represents a stalk which, emerging 
 from the materia alba or heap, was extending horizontally, and 
 changed abruptly for an upward direction, with the upper part 
 bearing the cluster. It is seen much aslant, but fortunately in- 
 tact (stained with acidulated solution of iodine, and magnified 
 to 1,700 diameters); it has a round end, with a fine rosette 
 of six rays, formed by six terminal sterigmata, and having the 
 six last spores, probably yet unripe, to their tops. 
 
 Fructifications by Temporary Spores (Sporidsl) and 
 BY Persistent Spores (Teleutosporesl). 
 
 The mycetologists and the algologists call temporary spores, 
 agamic spores, sporids, and conids the spores which are formed 
 without previous fecundation, and are intended by the multiplica- 
 tion of the species in more favourable and immediate conditions 
 (namely, the conids of the Peronospora, destined to diffuse the 
 species during a part of the year) ; whilst the persistent winterly 
 spores, oospores, or teleutospores, are produced by the act of con- 
 jugation, and help to preserve the species from external injuries, 
 and to strengthen their future shoots (like the hybernating spores 
 of the Feronospora in the thickness of the hospitable parenchyma, 
 deputed to reproduce the species in the following year).- Of the 
 two processes of sporification, one (says De Bary) seems intent 
 upon preserving intensively the species through conjugation, the 
 other only to increase its extension* 
 
 De Bary, Du Diveloppement des champignons parasUaires (Ann, des 
 Scien. Nat. Sir. Bot., t. XX.). 
 
AND CONTENTS OF THE MOUTH. 173 
 
 Of the other processes of propagation through gemmules, 
 sprouts, etc., we have already treated in the previous Memoir, and 
 shall refer to the subject again later on. 
 
 All the fructifications of our parasite, hitherto described and 
 drawn, probably belong to the temporary series, with the excep- 
 tion of the specimen given in Fig. 13 of the preceding Memoir, 
 which might be included in the persistent series ; but from the 
 last researches and other isolated cases, of which we will speak 
 presently, it appears that the same parasite presents also a com- 
 paratively scanty number of fructifications of another kind, which, 
 although similar in shape or type, assume special characteristics, so 
 that we are rather inclined to refer them to the persistent series. 
 
 We have already mentioned the club-shaped stems, two of 
 which, stained with gentian violet, are seen drawn in Fig. 26, 
 magnified to 1700 diameters. The club is generally long enough, 
 as in a j but sometimes we meet short ones, as in b, which might 
 be called an incipient phase. In the first named specimen, the 
 club, although complete, is still quite bare, and, to all appearances, 
 represents a hardly-formed expansion, before the sterigmata and 
 the spores have germinated. In fact, in the Fig. 2 7 (same staining 
 and magnification) may be noticed an ear, on the whole larger 
 and with spores proportionally more conspicuous, exhibiting an 
 internal stem, club-shaped, and brilliantly coloured, quite dissim- 
 ilar from the pale and slender stems of the first series, but 
 analogous to the bare stem of Fig. 26. 
 
 To our knowledge, these ears never reach the length of some 
 others having slender stems (see Figs. 12 and 16), which may 
 depend upon the comparatively limited length of the club-shaped 
 expansions. The spores of such ears are, besides, more conspic- 
 uous and pressed together, so as to form on both sides a sort of 
 zone or violet aureola, at a Httle distance from the stem ; and, 
 between this and the periphery, runs a clearer intermediate zone, 
 where the viscid substance is less coloured, but yet capable of 
 disguising the sterigmata. The light proceeding from the con- 
 denser must, in fact, cross first the deep violet zone, which 
 is next the slide, then the intermediate clearer substance (the 
 index of refraction of which is identical perhaps with that of the 
 peduncles), and finally the opaque violet zone, which overlooks 
 
174 BACTERIA OF THE SPUTA 
 
 the cover-glass. At any rate, the result of this optical combination 
 is to hide the peduncles. When in the solution of iodine we come 
 across such ears, the zones become mixed up, and we perceive, on 
 the whole, a triplex series of coloured granules, as is shown in 
 Fig. 13 of the previous Memoir, incompletely represented, which 
 would lead to the supposition that in these ears the secretion of 
 glair is more abundant and thick.* 
 
 It appears, besides, that such ears are even more compact and 
 resisting to the mechanical agents of disintegration ; also, their 
 fragments always exhibit a cohesion of the single particles,, and 
 their brilliant colouring becomes more conspicuous with aniline. 
 We remember having often found similar fragments in sputa ; but, 
 not then knowing their nature, we overlooked them. We also 
 remember that some sound ears, or fragments of the same, were 
 found mixed with many minute ears in fructifications upon small 
 flakes of urethral mucus, as we mentioned in the previous Memoir. 
 We may, however, state that, in the preparations of the dental 
 patina, ears of this sort are scanty in comparison with those of 
 the preceding form. 
 
 In considering now those more robust and conspicuous forms 
 of fructification, the mind tries, through analogy, to connect them 
 with the process of fecundation, and finds, although indirectly, its 
 existence is confirmed. In the preceding Memoir, we have des- 
 cribed some pseudo-inflorescences in tufts, having points varying 
 in shape and size, which we held to be future spindle-like bacilli 
 {^Bacillus tremulus of Rappin) and future comma, or serpentine 
 bacilli, destined, after being dissevered from the stem-and becoming 
 free, to perform the functions of sperraatia or antherbzoids. We 
 gave the reason for such hypothesis, as we also pointed out the 
 likeness of those elements (supposed male organs), with the sper-- 
 matia of certain well-known fungi, like Sphcerella sentina, Fumago 
 salicina, Apiosporium. citri, etc. 
 
 The antherozoids or spermatozoids in sea weeds, and thejr/«'w«- 
 tia in fungi were considered as elements of fertilisation. The 
 first {migratory filaments, spiral filaments, seminal corpuscles^, now 
 
 * After presenting this Memoir, we have made further researches (especi- 
 ally on the presence of such ears in the sputum of pneumonia, and on the manner 
 of detecting the peduncles), which we will soon make known. 
 
AND CONTENTS OF THE MOUTH, 176 
 
 cylindrical, now ribbon-shaped, furnished with cilia and endowed 
 with spiral movement in various directions, are originally contained 
 in a cellule or male organ {antheridium). The spermatia cor- 
 puscles, oval or in rods, straight or curved, also very motile, like 
 the analogous forms of the mouth, were, nevertheless, held to have 
 no cilia. They are originally sometimes contained in an appro- ■ 
 priate cellule or male organ {spermogoniuni), sometimes they grow 
 freely on the apex of the filaments, and get dissevered simply 
 through disjunction. In our parasite we tho'ught, at first, that the 
 fertilising elements belonged to this last type, and were formed in 
 a free state on the stems ; but we shall see, by and by, that perhaps 
 even they originate within apposite sheaths, and therefore may be 
 referred to the first type. In general, the spermatia, unable to 
 multiply through fission, have been seen, at times, to germinate 
 on their own account ; one common example of this kind is 
 exhibited in the ergot. 
 
 Now, we repeat, the existence of fructifications more conspic- 
 uous and distinct from the others (through their large club-like 
 stem and their two zones of colouring, etc.), in the normal parasite 
 of the mouth, would be quite explained, admitting them to possess 
 fertilising elements constituted by spindle-like, comma, and serpen- 
 tine bacilli, already described by us, and holding the other spores 
 as agamous and temporary. Perhaps the persistent spores in 
 this parasite are destined to go through the intestinal tube unin- 
 jured, withstanding the dissolving action of the gastric juices, and 
 emerge into the external world, maintaining in the faeces their 
 vitality for the future diffusion of the species. 
 
 ■ As regards the function of conjugation, it may be performed 
 on the already formed filaments, as we see in many other crypto- 
 gams, where sometimes the act takes place between two contiguous 
 filaments, the male organ of the one penetrating the female organ 
 of the other ; but nothing prevents us from beUeving that a ferti- 
 lisation of another kind may have taken place between the male 
 element (spindle-like, comma, or serpentine bacillus) and the 
 mother spore, before the germination of the fertile filament. 
 
 Against these views of ours, a quite opposite hypothesis might 
 be produced, namely, the hypothesis of a commensalism or sym- 
 biosis. In such hypothesis the small sporules, and especially the 
 
176 BACTERIA OF THE SPUTA 
 
 productions by points, would not be proper phases of Leptothnx-, 
 but parasites oi the parasite, or new micro-organisms of another 
 species, come to implant themselves and thrive at the expense of 
 the original parasite in the same way as Leptothrix parasitica, 
 Kiitzing, which with its slender filaments lodges itself on the larger 
 filaments of Zygnema and Cladothrix dichotoma, as we see in 
 Fig. 2 1 (stained with vesuvine and methyl violet, then with solution 
 of iodine, magnified to 600 diameters). But, in this way, one 
 might object, for argument's sake, that grapes are so many para- 
 sites of the vine on which they fructify. In fact, consulting the figure 
 in question, anyone can see that the secondary parasitical shoots, 
 c, c, are less thick than our points, which engraft themselves round 
 the stem, like the hairs of a bottle brush. They are not, besides, 
 methodically arranged, but stretch out very much, like stems des- 
 tined to vegetate on their own account, rather than to complete 
 the organism bearing them. The filaments of Leptothrix parasitica 
 implant themselves also upon the stem of Zygnema, a, a, or of 
 Cladothrix, b, b, which feeds them, by means of bulbs, or spores 
 originated in them, s, s, as the Fig. 22 shows still better (same 
 staining, magnified to 1,600 diameters) : spores which are not 
 seen at all at the insertion of our points. And still less the fila- 
 ments of Leptothrix parasitica are seen to drop at last from the 
 central stem, and swim in the medium with the same briskness of 
 our spindle-like, comma, or serpentine bacilli. 
 
 Various Aspects and Forms. 
 
 In the preceding Memoir we have spoken of various forms and 
 appurtenances of the parasite .in question ; we specially point out 
 the bifurcations and trifurcations towards the seat of certain fila- 
 ments, with tiny radical swellings, like haustoria; and then the 
 more pronounced ones, some at the knots, some at the apex : the 
 latter like small heads. We have already mentioned the other 
 apical swellings (fertile filaments), club-shaped. 
 
 We now go on to describe a third form of apical expansions, 
 very scarce, which, provisionally, we shall denominate sheath 
 expansion. 
 
 Such an expansion, represented in Fig. 28 (stained with gentian 
 violet, magnified to 3,100 diameters), is very pale, has streaks in 
 
AND CONTENTS OF THE MOUTH. 177 
 
 its contour, not detected with inferior objectives. Its paleness 
 cannot be attributed to insufificient colouring, because the examined 
 form rose on the top of a filament (likewise pale) in the midst of a 
 thick and very pretty tuft of ears brilliantly stained (of the 
 kind shown at Fig. 12), and nearly surpassed them with its 
 point. The external contour is very slender from the base of the 
 expansion up to its point, and between the internal stem and the 
 exterior contour are seen numerous slender, tiny, transverse 
 threads, which are attached to the external sheath by means of 
 more prominent small dots. 
 
 At first we could not understand the probable meaning of this 
 structure. That it was not to be taken for the club-like expansions , 
 of Fig. 26 was evident from its paleness, being the antipodes of 
 the bright colouring of these, as well as the presence of the inter- 
 nal fine beams. On the contrary, after deeper reflection, we 
 thought we might refer it to other forms already described in the 
 previous Memoirs. One of these forms would be that of the pseudo 
 inflorescences in tuffs (preceding Memoir, Fig. 14). Comparing 
 the two figures, it will not appear unlikely that the slender threads 
 of Fig. 28 in the present Plate, distended like fine beams between 
 , the internal stem and the sheath, growing more and more in a 
 transverse direction, may end by breaking the external envelope and 
 become ix&e points, perfect fertilising elements (spindle-like, comma, 
 or serpentine bacilli), at first only free from the surrounding 
 sheath, in order to constitute the tufts of the preceding Plate, and 
 at last becoming disjoined from the central stem, so that they may 
 fulfil their function through that stirring motion, mentioned in the 
 previous work. 
 
 The other form, possibly analogous, would be that delineated 
 in the first Memoir (Fig. 2, n, lower down), similar to a large 
 bacillus, singularly veined throughout, and for a certain tract 
 having traces of lineal bacilli. Even this is a rather rare form, 
 and we have never found it on the top of any filament, but quite 
 by itself. We have already hinted in the previous Memoir that 
 ^ that veined bacillus might be a sort of receptacle for the future 
 fertilising elements, as an antherid or a spermogone. 
 
 Now, supposing that interpretation true, the transmutation from 
 
178 BACTERIA. OF THE SPUTA 
 
 one to the other of the three forms would become clear enough. 
 We should have, in the first form (or sheath expansion) of Fig. 28 
 - (present Plate), an antherid or sperniogone, hardly shown, 
 and in the second form (the tufts of Fig. 14, previous Memoir) a 
 male organ in full development. The third form (the veined 
 bacillus of Fig. 2, n, below, first Memoir) would be an inter- 
 mediate form to the other two (an arrested form), or an antherid 
 or spermogone, prematurely fallen from its stem, having been 
 unable to attain to the adult form of tuft ; and then strayed from 
 its destination and remained, as it were, unripe, or even returned^ 
 to a neutral condition, which is, we think, common to the severed 
 or truncated filaments, as, having been unable to attain the fructi- 
 fication, they limit themselves to a reproductive function of a 
 lower degree (fissiparous multiplication) through the increase of 
 the granules or the lineal minute elements, contained in the 
 interior of their sheaths. If this were confirmed by farther 
 researches, it would lead us to rectify the first supposition about 
 the formation of the fertilising elements. They would not form 
 themselves freely on the top of the respective filaments, but within 
 a receptacle or male organ properly so called (sheath expansion). 
 
 These are only simple conjectures, aiming at connecting the 
 various forms hitherto described, reconducting the appurtenances 
 of our parasite to the general laws of the cryptogamic flora, and 
 far from pretending to give herewith a full and exact explanation of 
 them. We shall be quite satisfied if the features of the facts we 
 have endeavoured to describe can be proved by further researches. 
 
 But, even upon a simple descriptive ground, we should perhaps 
 overstep the limits of a simple preliminary study if we were to 
 dilate longer in the investigation of other particulars, before seeing 
 confirmed and set up the points already demonstrated (such being 
 the most important) by competent authorities. Neither is it our 
 business to solve the question whether the discussed parasite is a 
 fungus or an alga. We shall only say that it appears to us to 
 partake of the characters of both families, to thrive as an alga, 
 but to fructify similarly to certain fungi. We shall, therefore, 
 limit our remarks about some apparent irregularities, in the aspect 
 of the described ears, which, through inattention, might pass for 
 true irregularities or anomalies of structure. 
 
AND CONTENTS OF THE MOUTH. 179 
 
 In the first place, we refer to some gibbosities or irregular pro- 
 jections, which are met sometimes by the side or upon some ears, 
 which might lead to the belief that the ears themselves are perhaps 
 constituted without any order, or that the series or longitudinal 
 rows of the sporules are not always six in number, but at times 
 more. 
 
 Now, one of the more frequent causes of such irregular appear- 
 ance is very simple. The breaking up of a contiguous ear, and 
 the adherence of that extraneous fragment to the ear that we 
 examine ; having frequently verified this occurrence, we considered 
 it superfluous to draw a similar specimen on our plate. The 
 other case, less frequent, is drawn in Fig. 29 (saturated with 
 acidulated solution of iodine, magnified to 1,700 diameters). 
 Here it is not the superposition of an extraneous fragment, but 
 the folding up of the ear itself In the figure referred to, the third 
 superior and the point b of the ear are turned up and pressed 
 back upon the middle third, but in a direction somewhat oblique 
 to its axis, so that, at first sight, or with inferior objectives, they 
 simulate a gibbosity. But focussing with the fine adjustment, it is 
 easily perceived that there is, in reality, a folding due to a mechan- 
 ical cause, and accidentally rendered even more pronounced by 
 the pressure of the cover-glass. 
 
 We do not speak of a third apparent irregularity, which might 
 deceive us only when using inadequate optical means — we mean 
 the accidental apposition of extraneous cocci, of bacteria and comma 
 bacilli around some ears, as shown in Fig. 23, b, b. Under lower 
 power objectives, such cumuli or groups, especially if more con- 
 spicuous, may, in fact, simulate real protuberances. 
 
 There is, however, a special apposition of cocci and bacteria, 
 sometimes visible on certain ears ; perhaps, those remained longer 
 with their points in contact with the labial mucous, and there 
 became incrusted with the above bacterial elements, through a 
 cement of viscid mucus. Such ears, in fact, are never augmented 
 on the opposite side, but only on the top, like a very oblong pear ; 
 and the increase, gradually narrowing itself, seldom goes beyond 
 the half of their length (Fig. 30, stained with gentian violet, mag- 
 nified to 1,170 diameters). 
 
180 BACTERIA OF THE SPUTA 
 
 When they reach the first colourising stage with the gentian 
 violet, we can easily perceive there is in reality a sort of cap (at 
 first, more pale and granular) constituted by cocci and bacteria, 
 only slightly coloured, in various layers, towards the point a, and 
 sloping towards the half, a', a' ; whilst the ear, b, b, with its sporules, 
 is seen brightly coloured in the interior. In the second stage, 
 even these adventitious cocci and bacteria become coloured, and 
 the cap is no longer distinguished from the internal ear. 
 
 CONCLUSION. 
 
 We have seen how Billet describes the evolutionary cycle of 
 the Bacteriaceae (which would constitute for our parasite only the 
 inferior cycle), and that his remarks mostly agree with those we 
 have made on the same parasite. His interpretations of the 
 various phases of that cycle do not differ from ours, excepting in 
 what affects the production of bacteria included in filaments, 
 which Billet considers are real endogenous spores ; and we cannot 
 positively deny that such is the case in the four species studied by 
 him ; whilst in our parasite they ought, in our opinion, to be held 
 as simple gemmules of reserve. 
 
 But from the exposition of the facts in our previous Memoir, 
 and confirmed in the present one, it clearly results that the evolu- 
 tionary cycle, so nicely delineated by Billet, cannot include all the 
 morphological phases of our Leptothrix racemosa, but only some of 
 them. In this parasite, besides that first cycle which we call 
 inferior, there is another — the superior, which comprises the 
 organs of genuine reproduction and fructification. Finally, toge- 
 ther with these two normal evolutionary cycles, there is another 
 one- — accidental, called virulent, in which (according to laboratory 
 experiments) it seems that certain elements, derived from the 
 parasite itself, may, as Pommay says, develop themselves in the 
 sense of virulence. 
 
 The first two cycles would, therefore, constitute the morpholo-^ 
 gical series, and the third, or virulent, cycle would, in modern 
 language, constitute the biological series of our parasite. 
 
 Morphological Series. — Inferior Cycle. — The inferior cycle 
 embraces the following phases : — ' 
 
AND CONTENTS OF THE MOUTH. 181 
 
 I. — Phase of Vegetation. — The characters of this phase are 
 those assigned by Billet to the filamentous state, only that he con- 
 siders the intertwined state {Hat enchevUrk) as a distinct and later 
 phase ; whilst we believe that it, in our parasite, accompanies the 
 filamentous state, in the same manner as the mycelium or creeping 
 vegetation in fungi. In other words, the intertwined state is even 
 posterior to the isolated filaments which lead a wandering life in the 
 liquid substrata, being unable to attain a more vigorous and stable 
 one ; but it does not constitute a distinct phase, as it is quite natural 
 that, when the passage to the superior phases is precluded /a^r/^/ 
 vegetation) for want of a fit soil, the filaments intertwine and drop 
 to the bottom, without being able to spread like the mycelium of 
 fungi. However, under favourable conditions, a kind of mycelium 
 (a more complete phase of the entangled state) may be formed, 
 giving birth to an aerial vegetation, as may be specially observed 
 in the patina of the tongue and in the deep strata of the patina 
 dentaria. 
 
 II. — Phase of internal gemmulation or budding and dissemina- 
 tion. This partly corresponds to the dissociated state of Billet, 
 through the disarticulation of the knots of the single filaments, or 
 through setting free the included bacteria ; and it is clear that the 
 unstable condition of the nutrient medium continuing, and conse- 
 quently the passage to the superior phases being prevented, no 
 other way of perpetuating the species is left to these micro-organ- 
 isms than the inferior reproduction or simple multiplication by 
 shoots and gemmules. Looked at in this way, this phase would 
 even comprise that held by Billet as endogenous sporulation, but 
 which are, at least in our parasite, gemmules of reserve, properly 
 destined to the multiplication of the species, in a neuter state, 
 through simple fission of the elements, when the genuine repro- 
 duction, by means of seminules or spores, is not possible, or when 
 the fertile filaments have been dissevered or cut by mechanical 
 injuries. 
 
 III. — Protective phase. This phase fully corresponds to the 
 zoogloeic state and to certain conditions of the dissociated state of 
 Billet, as we have pointed out in the first paragraph. The presence 
 of su6h forms has been undoubtedly detected in the mouth, and 
 
182 BACTERIA OF THE SPUTA 
 
 even within the relative epithelia. They would appear to be a 
 kind of reserved fund, preserved in case of any alteration of the 
 future conditions of pabulum and surroundings, as the author 
 properly says. We shall speak by-and-by of the relationship of 
 this phase with the diplococcus of pneumonia. 
 
 Here Billet would end the evolutionary cycle ; for us, on the 
 contrary, these first phases would only constitute a cycle, at times 
 preliminary, at times succedaneous to the second or superior cycle. 
 The varied elements of the first cycle, being taken separately and 
 held as special beings or complete living individuals (filaments, 
 bacilli, bacteria, and cocci), are in reality only particles, trunks, 
 organs, series of cellules, or cellules endowed mostly with fissipa- 
 rous multiplication, but destined to constitute a more complex 
 organism. 
 
 Superior Cycle. — This comprises the phases of the properly 
 called life of reproduction, and these phases are three. 
 
 IV. — Agamous Fructification. — Referring to the general laws 
 of multiplication of the phanerogams by means of bulbs, tubers, 
 shoots, or buds, and of their genuine reproduction by means of 
 seeds, it should be borne in mind that with transplantation are 
 transmitted the accidental modifications brought about by domes- 
 tication, grafting, etc., which, in the long run, may end in the 
 degeneration of the plant ; whilst with seeds, on the contrary, the 
 species reverts to the natural vigour of the wild state. 
 
 Now, we incline to believe that something similar may happen 
 even with cryptogams. Probably in those which have, besides a 
 fissiparous multiplication, a true reproduction through spores, the 
 sporulation will mean that the species resumes its native vigour, in 
 spite of the attacking or enfeebling causes which, as in our para- 
 site, may impair its vigour. 
 
 The seminule or spore may germinate without previous ferti- 
 lisation, and (as we have already said) we believe this may happen 
 more frequently in our parasite. In the species studied by Billet, 
 this external sporification or fructification was wanting, and it is 
 remarkable that the cocci, which are found abundantly in the 
 buccal contents, were hkewise wanting. It remains to be seen 
 whether this want of fructification is really the rule, or simply a 
 
AND CONTENTS OF THE MOUTH. 183 
 
 consequence of the nature of its pabulum and the material upon 
 which Billet based his researches. Under other conditions, upon 
 media not only stable, but favourable to the production of a myce- 
 lium-like growth, a true and proper fructification mighty even in 
 the above-named species, take place. 
 
 In ^ur parasite, the conditions indispensable for the fructifica- 
 tion are : — a, The solidity and nature of the soil; b, the protection 
 against attrition ; and c, moisture of the saliva. 
 
 V. — Organs and Fertilising Elements. — By the side of the 
 agamous sporulation we have, in many cryptogams, that of conju- 
 gation. This admits of male organs and fertilising elements or 
 spermatic threads, of which we have already spoken. In our 
 parasite, likewise, we have male organs and fertilising elements. 
 The male organs would first show themselves in the state of young 
 sperraogones or antheridia (or of spermogones or aborted anther- 
 idia, not developed or reverted to the neuter state), and afterwards 
 in the state of tufts of ripe pseudo-inflorescences, proceeding from 
 the first type. On the other hand, the fertilising elements would 
 be constituted by the spindle-like, comma, or serpentine bacilli, 
 already formed in the described organs, and finally set free, through 
 the disarticulation of the tufts already mentioned. 
 
 VI. — Conjugated Fructification — i.e., that with clavated stem, a 
 dual zone of colourisation, clusters, or more conspicuous ears, 
 more bulky and compact spores, destined, perhaps, to cross unin- 
 jured the alimentary canal, and to remain alive in the faeces, with- 
 standing the dissolving action of the gastro-intestinal juices. 
 
 Nature, as we see^ has been prodigal to this parasite, by its 
 various manners of multiplication, adapting each of them to this 
 or that condition of the nutrient substratum, in order to preserve 
 and multiply its species, in the midst of numberless and very 
 varied difficulties. As soon as a higher phase is precluded, or a 
 nobler element is thrown back, by external injuries, to an inferior 
 degree, it does not stop from disseminating everywhere particles 
 apt to germinate and spread extensively the species when it cannot 
 do it intensively. We are reminded that Nature has even wished 
 to endow this tiny plant with such a tenacious life, that its elements 
 on the human teeth cannot be destroyed for centuries, as exhibited 
 
184 BACTERIA OF THE SPUTA 
 
 by the tartar on the teeth of Egyptian mummies (vide the preced- 
 ing Memoir). 
 
 If here ends, at least provisionally, the Morphological Series of 
 our parasite, it only remains to us to say two words on the Biolo- 
 gical Series in the modern sense, or, rather, on the pathological 
 phenomena, assigned to some of its forms, or to specific bacteria 
 similar to the latter. 
 
 Biological Series. — Under this title we comprise but three 
 forms : the Fneumococcus, the Bacillus of Koch, and the Gonococcus 
 of Neisser, already demonstrated, according to our views, in the 
 two previous Memoirs. 
 
 I. — Fneumococcus. Few now hold that the pneumococcus is a 
 specific bacterium, arising externally. It is generally considered 
 to be either an habitual germ of the mouth (Micrococcus of salivary 
 septicosmia), or, in common with Pommay, a saprophytic bacterium 
 evolving itself in the sense of virulence. Perhaps it will in time 
 be known as a simple dissemination of the zoogloeic phase of our 
 parasite, following on the formation of the pulmonitic exudation, 
 as its greater abundance in the last stages of pneumonia, its 
 presence in traumatic pneumonia, and other evidences will 
 prove it to be so. But even admitting that the evolution in the 
 sense of virulence may take place in the mouth and not in the 
 respiratory organs affected (as regards virulence, which may be 
 inocdlated) ; admitting that it has preceded and not followed 
 pneumonia, we cannot necessarily infer that the disease proceeded 
 from this coccus. But even the colonies of these diplococci, when 
 repeatedly transplanted in other culture media, or even the media 
 themselves, may become contaminating through inoculation, this 
 circumstance may also occur with other salivary bacteria , (for 
 example. Bacillus crassus sputigenus), and might be interesting in 
 experimental pathology ; but, botanically speaking, it does not 
 implicitly imply a separate specific entity. We repeat that we do 
 not impugn, but rather try to conciliate the results of the inocula- 
 tions with those of the morphological research. 
 
 II. — Bacillus of Koch. Our remarks concerning this bacillus 
 are very similar, and we hope at a future time to devote a special 
 Memoir to it. The reasons which induced us to believe the 
 
AND CONTENTS OF THE MOUTH. 185 
 
 bacillus in rosaries to be a dissemination of the small spores of 
 Leptothrix spread over the tubercular spots, and the rod-shaped 
 bacilli as proceeding from other elements of Leptothrix, have been 
 already given in the preceding Memoirs. Their greater dissemi- 
 nation in phthisis than under other conditions may depend upon 
 different causes, and partly, perhaps, from the following simple 
 reason : — Breathing through the nose, as we generally do, only a 
 few germs of a purely buccal origin are inhaled ; but the position 
 becomes altered when we breathe through the mouth as well, as 
 happens in phthisis, bdcause of panting or burning heat :— 
 
 " . . open'd wide his lips. 
 Gasping as in the hectic man for drought. 
 One towards the chin, the other upward curl'd." 
 
 —Dante, Inferno, xxx., Gary's Transl. 
 
 Now, the small sporules of Leptothrix exclusively originate 
 from the patina dentaria, and without that particular form of 
 breathing {i.e., by the mouth) they cannot gather in any great 
 number into the air-passages. Many also, in sleeping, breathe 
 likewise with their mouth ; but in the normal conditions, however, 
 the ciliary action continuing unhurt through the air-passages, the 
 inhaled leptothrical elements cannot reach the pulmonary tissue, 
 but are instead thrown out and expectorated with the sputa, as 
 stated in the other Memoir. 
 
 Ill, — Gonococcus of Neisser. Probably, says Pommay, a normal 
 bacterium of the urethra is the progenitor of the gonococcus of 
 Neisser; it is useless to repeat here that reMtionship. Let it 
 suffice to record the fact that we have detected in the urine num- 
 berless gonococci, in a case of carcinoma of the bladder, inde- 
 pendently from any gonorrheal contagium whatever.* According 
 to the morphological characteristics, the gonococci in question do 
 not differ from the ordinary arched diplococci, and therefore can 
 be held, till proved otherwise, to be derivations from the state of 
 sarcina (one form of the zoogloeic state). As regards the manner 
 by which the leptothrical elements penetrate from the external 
 genitals into the urinary passages, as well as the presence of analo- 
 *AtH delta R. Accademia Medico-Chirurgica di Napoli, tomo XLiil., 1890. 
 
186 BACTERIA OF THE SPUTA, ETC. 
 
 gous forms (curved diplococci) in the contents of the mouth, in 
 sputa or the middle ear ; and also as regards the discovery in the 
 urine or sperm of capsulated forms (forms analogous to pneumo- 
 cocci), we refer to the previous Memoirs. 
 
 From the morphological sketch we have just given, nothing 
 perfectly absolute and incontroverted is to be inferred, either for 
 or against any previously acknowledged systematic view; but 
 perhaps few people will doubt that, in the field of modern bacte- 
 riology, much still remains to be done, and a great deal of discus- 
 sion, combined with careful study, will be required before any 
 definite and satisfactory results will be arrived at concerning the 
 pathogenesis of infectious diseases (we are, of course, now alluding 
 to those cases which are strictly clinical). Many will, we are sure, 
 agree with us that, with regard to bacteria, our knowledge is so 
 limited that no method of investigation will appear superfluous in 
 order to study aright their genesis and properties in order tho- 
 roughly to understand and appreciate their function in our 
 economy. 
 
 F. ViCENTiNi, Corresponding Member. 
 Chieti; May, i8g2. 
 
Fig. 31. 
 
 X 400. 
 
 Fi6. 33. 
 
[ 187 ] 
 
 Bacteria of tbe Sputa ant) (Tri^ptoaamic 
 flora of tbe flDoutb. 
 
 APPENDIX TO THE ENGLISH EDITION 
 Translated by E. S. 
 
 Summary. 
 Reasons for writing this Appendix. Doubts about the Bacte- 
 rium hypothesis (the views oi Jaccoud, Middendorp, Petten- 
 kofer, and others). Hypothesis of the author on the action 
 of Bacteria upon the human and animal economy (in the 
 normal state, in disease, in labdratory experiments). Some 
 further remarks on the morphology of Leptothrix. 
 
 Reasons for Writing this Appendix. 
 
 IN making this translation, I was, at first, tempted to recast the 
 preceding Memoirs into a single work, shorter and more 
 synthetical ; but I have chosen to re-introduce them in their 
 original form and arrangement, wishing to keep unchanged the 
 itinerary, so to speak, in stages, which I have followed up without a 
 preconceived plan, but simply guided by my own clinical investiga- 
 tions. In the present Appendix I shall endeavour to answer, as 
 briefly as possible, a few objections, and give some more particu- 
 lars on the morphology of Leptothrix. 
 
 The control exercised by other investigators upon the morpho- 
 logical phases described by me will be confined to three distinct 
 objects : — 
 
 I. — If my observations are exact, Leptothrix buccalis possesses 
 real organs of fructification, and probably also organs of fecunda- 
 tion, similarly to a fungus or an alga of a more complicated 
 structure. Consequently, the first point of control will be to 
 ascertain the existence of such organs. From the preface, which 
 Prof. Miller has kindly prepared for this edition, it is apparent that 
 he has had no great difficulty in finding the " grape bunches " 
 appearances described by me. Of such forms he drew the photo- 
 micrograph, reproduced in Fig. 31 (magnified to 400 diameters). 
 
188 BACTERIA OF THE SPUTA 
 
 Likewise, they have been detected by Mr. J. H. Mummery. In 
 his paper before the Odontological Society he says : — " 1 certainly 
 detected appearances very similar to those organs which Dr. 
 Vicentini compares to grape bunches, and made drawings of 
 them at the time. The regular lines of rounded bodies, which 
 the author considers to be spores, were plainly seen, as well as the 
 central stem ; but I was unable to see the minute peduncles, by 
 which he describes the spores as attached to the central stem. 
 The club-shaped appearances of these bodies were also very 
 evident ; but so far I have been unable to see anything that I can 
 identify as corresponding to two forms of the male organs, although 
 I do not consider I have done justice to the author in my search 
 for these, and shall renew the attempt. The forms which I saw 
 certainly seem to be too regular in shape to be explained as an 
 accidental accumulation of granules or micrococci around an 
 isolated filament (an explanation which has been given by some 
 observers)." * 
 
 Of the existence of forms by ears or grape bunches there 
 appears, then, to be no doubt ; it remains only to be seen' if, as I 
 detected with a i/asth inch immersion lens in such ears, real 
 peduncles or sterigmata exist, and whether the other forms, con- 
 sidered by me as male organs or fecundating elements (not to 
 speak of less important forms) are as I described them. 
 
 2. — Let this first question, relating to facts, be once solved, 
 and there naturally arises that of their interpretation. Are the 
 above-named ears true fructifications and the tufts real male 
 organs ; or is it, in both cases, a question of commensalism or sym- 
 biosis, or some other phenomenon ? 
 
 Some correspondents have expressed to me their doubt that 
 the ears in question might only be a particular zooglceic form grow- 
 ing into the internal dental tubules, already described by Zopf.f 
 
 But everybody can see that between the two there is not a 
 single point of analogy in the structure. The figure of Zopf, 
 
 *J. H. Mummery, Transactions of the Odontological Society of Great 
 Britain, Jan., 1894, p. 77, partly reprinted in the Lancet Feb. 24th, 1894, 
 Vol. I., pp. 487—88. 
 
 tZopf, of. cit.. Fig. 4, Miller, "Der Einfluss der Mikroorg. a. d. Caries," 
 etc. (Klebs' Archiv, XVI., 1882, Fig. 7). 
 
AND CONTENTS OF THE MOUTH. 189 
 
 reproduced by Miller in KleM Archives, represents a simple heap 
 of cocci and bacteria, mechanically modelled within the dental 
 tubules, and not a production germinating upon \he.free surface of 
 the teeth. It has no central stem, sterigmata, or peduncles ; its 
 single element's are heteromorphous, having at one end bacilli, in 
 the middle bacteria, and at the other end cocci ; whilst the ears 
 bear round, uniform spores in six longitudinal rows. Finally, the 
 heaped-up elements in the dental tubules are more bulky and 
 easily discernible in their normal state. On the other hand, the 
 spores in question are very much more minute, and cannot be 
 distinctly detected without being properly stained. 
 
 With regard to the process of eventual fecundation, which I 
 have foreseen in the Leptothrix, it will not be inopportune to 
 repeat what Dr. Dallinger writes on the possible conjugation of 
 bacteria : — " Nothing like ' conjugation,' or any other form of 
 sexual generation, has yet been witnessed in any bacteria; and 
 until such shall have been discovered, no confidence can be felt 
 that we know the entire life-history of any type. As it seems 
 unquestionable that among the higher fungi ' conjugation ' often 
 takes place at a very early stage of growth, it seems a not very 
 improbable surmise that the 'granular spheres ' observed by Prof. 
 E. Ray Lankester, in his Bad. Rubescens, may be a product of 
 conjugation in the micrococcus stage of these organisms." * 
 
 3. — Having solved these two questions (namely, of the facts 
 and their interpretation), we may with some reason enter the field 
 of the third question, or of the deductions to be derived from the 
 exhibited facts, concerning the single isolated bacterial forms of 
 the buccal cavity, and see whether such forms really constitute as 
 many independent species, or if many of them are not rather 
 simple disseminations or rudiments of one or more superior species 
 of micro-organisms. 
 
 Now, it so happens that in looking at my researches this last 
 part has had the first consideration, particularly in relation to 
 pathogenic bacteria. Prof. Miller, besides, attributes to me as a 
 firm and full conviction what I simply suggested as doubtful and 
 
 ♦Carpenter, The Microscope and iU Revelations, 7th edit., by Dallinger. 
 (London, 1891, p. 588.) 
 
190 BACTERIA OF THE SPUTA 
 
 hypothetical— viz., that a great proportion of the bacteria, which is 
 prolific in the mucous membranes, might proceed from a common 
 stock. But I emphatically state that a definite opinion upon that 
 subject would at the present moment be premature. 
 
 Prof Miller and Mr. Mummery hint at the 'revolutionary 
 character of my views ; but others, long before me, had advanced 
 the problem of an eventual simplification of the bacterial species. 
 The opinions of Hallier and others upon this subject are well 
 known, and Mr. Mummery has opportunely recalled the opinion 
 of Naegeli, as well as a passage in conformity with the same, from 
 the article, " Fungi " of the Encyclopedia Britannica* 
 
 Finally, we must repeat the hypothesis of Brefeld concerning a 
 possible derivation of bacteria irom fungi of a higher order, which 
 may have lost their organs of fructification, so that their successors 
 could multiply by no other manner than by fission. + 
 
 My reason for pointing out the problem of the classification of 
 the species of bacteria was simply to anticipate the objections of 
 specialists against ray experiments, and I will now endeavour to 
 explain away some of those objections. On this point Mr. 
 Mummery recalls the fact advanced by Prof. Miller, that " a pure 
 culture of the micrococcus of the sputum septicaemia, one or two 
 days old, will invariably kill a mouse or a rabbit ; whereas a pure 
 culture of some other bacterium from the same mouth, under 
 exactly the same conditions, may have no action at all. This (he 
 adds) seems very extraordinary if they are all derived from one 
 single form in the mouth." % 
 
 We might admit a distinct specific entity for this or another 
 definite form of bacteria decidedly virulent, without admitting the 
 specific entity of all the remaining forms ; but in the second Memoir 
 I have already referred to the insufficiency of peculiar vital action as 
 forming a basis for the classification of the bacterial species. The 
 distinct characteristic of the other cryptogamic species (I said) is to 
 be found in the fructification or the completed series of the various 
 
 * Mummery, loc. cit., pp. 79, 80. 
 
 t Brefeld, Untersuch. U. d. Spaltpihe, Bac. Subtilis. ( Gesell. Nat. Freunde, 
 Berlin, 1878, Schimmelpilze, Heft. 4). 
 
 X Mummery, loc. cit., p. 80. 
 
AND CONTENTS OF THE MOUTH. 191 
 
 morphological phases of each. No one would pretend to classify an 
 alga and a. fungus from observation based upon experiments of the 
 vital action of their single elements on their hosts. But, departing 
 from the ordinary rule, if we wish to assume such a distinct crite- 
 rium for the classification of certain species of bacteria, simply 
 because their natural history is at the present to a great extent 
 unknown, it does not follow that the same should be entirely 
 overlooked. 
 
 Many parasitic fungi, which in various ways infest their hosts, 
 assume different phases and forms, at times changing upon the 
 one or the other. These divers forms, formerly believed to be as 
 many species, are often simple modifications or particular condi- 
 tions of a single parasite, as happens for instance in the well-known 
 alternation of generations. 
 
 The alternation of generations (as, for example, in the Ure- 
 dinea, ' ergot,' etc.) exhibits many examples of this kind. Thus, 
 the Puccinia graminis, the cause of the mildew in wheat, only 
 represents a first phase (the teleutospore generation) of an alternating 
 fungus. The same fungus, passing afterwards from the wheat to 
 the barberry, undergoes a second phase {Mcidium Berberidis, or 
 the cecidiospore generation). From this a third generation is pro- 
 duced, which returns to infest the wheat, taking up then two dis- 
 tinct forms, one being a reversion to the primitive phase {Puccinia 
 graininis), ahd the other, altogether different, the Uredo form. 
 
 Further, the recent history of the Mycetology exhibits, at every 
 step, a progressive working of simplification in this sense. One 
 half, perhaps, of the old species of fungi has already disappeared ; 
 nor would it be surprising if a similar simplification happened even 
 for the bacteria forms. Are not bacteria even lower in the crypto- 
 gamic scale, more slender, and more exposed to the agents of 
 destruction ? No wonder, then, that they possess a greater faculty 
 of adaptation and a more extended polymorphism. 
 
 In the case of the micrococcus in the sputum septiccemia, in 
 spite of the virulence of its cultures, we are inclined to deny its 
 specific entity, and hold it to be a phase (a capsulated zoogloeic 
 form) of the Leptothrix itself considering its close analogy with 
 the proper capsulated forms of other and altogether different 
 species of bacteria — viz., Cladothrix dichotoma, Bacterium Bal- 
 
192 BACTEEIA OF THE SPUTA 
 
 Hani, Bacterium osteophilum, and Leptothrix parasitica (Kiitzing), 
 see Figs. 17 and 18. 
 
 Prof. Miller has, on this point, dissociated himself from a fact 
 apparently unimportant — viz., the change of state, a change which 
 by itself may, in certain cases, produce bio-physical and bio-che- 
 mical effects analogous to those of a real change of species. It 
 is, then, by overlooking the standard of the particular morphology, 
 that laboratory experiments may lead us erroneously to make an 
 unlimited number of species. 
 
 Consequently, the greater number, if not the whole, of the 
 isolated bacterial forms in the mouth could be related to Leptoth- 
 rix. If the minute cryptogamic plant which I described exists, 
 where shall we place its various particles, detached through ripe- 
 ness or removed by injuries (as spores, sporids, gemmules, chains, 
 filaments, enclosed bacteria, and male elements), if none of the 
 bacterial isolated forms belong to it ? 
 
 It is, I think, easier to suppose a common derivation of these 
 isolated forms from the above particles of the same plant (like 
 fruits or leaves fallen from the same tree) than the special and 
 altogether distinct origin of them from without. 
 
 With regard to the experiments of culture and inoculation, far 
 from minimising their importance, I intend to show later on that, 
 being considered within their proper limits, although interpreted 
 in a different way, such experiments may entirely coincide with my 
 opinion of a possible simplification of the bacterial species. True, 
 the bacterial science is still too modern to over-reach a whole 
 system of etiology, and generally the systems have never thoroughly 
 succeeded in medicine. Thinking of the limitation of our minds 
 before the mysteries of life, we may entertain the doubt as to 
 whether the intimate reason and essence of diseases in general, 
 and of the infectious ones in particular, might be accessible to our 
 knowledge, as the greater number of bacteriologists pretend. But, 
 however, I do not presume to minimise the importance of such 
 studies. I have already given in previous works the opinions of 
 several authors upon the limits to which the study of bacteria in 
 medicine may be applied. Now, I will add a few more recent 
 extracts, advising the reader to refer to the writings of Prof. De 
 
AND CONTENTS OF THE MOUTH. 193 
 
 Giovanni of Padua upon the relations which exist between the 
 laboratory and the clinic* 
 
 Doubts respecting the Bacterial Hypothesis. 
 
 /accoud's Views on the Spontaneity in Microbian Diseases. 
 
 Prof. Jaccoud in 1882 concluded one of his lectures in the fol- 
 lowing terms : — It is its source of origin, which makes the bacterium 
 infectious, and not any eflSciency attached to it simply as a bacte- 
 rium. The infectious properties of bacteria are properties borrowed 
 from the soil upon which they have lived, and which they can keep 
 up from generation to generation, even upon artificial cultures.t 
 
 He observed that the traditional empirical etiology of tubercu- 
 losis, as well as of other infectious diseases, maintains all its value 
 confronted with the bacterial doctrines ; that pneumonia, for 
 instance, and endocarditis are not always due to microbes coming 
 from without, but rather to an inward infection, following upon a 
 diminished resistance of the organism to microbes, which it bears 
 itself : indifferent pathogenic microbes, capable of provoking 
 affections of various seat and form. J 
 
 In 1888 he resumed the argument of such auto-infection, attri- 
 butmg it to pre-existing germs in the organism, harmless, but 
 which, owing to external circumstances (of a corporeal or cosmical 
 order) may become pathogenic.§ 
 
 In 1892 he deplored the tendency that modern bacteriology 
 had of suppressing medical etiology through three erroneous sup- 
 positions : — (i) By considering the pathogenic microbes as extra- 
 neous to a healthy organism ; (2) by considering them as the 
 
 * De Giovanni, sulla tisichezza pulmonare, Naples, 1882. Uno sguardo 
 alia batteriologia, 1886 — 87. La linfa di Koch, communicazione R. Istiluto 
 veneto, 1891, pp. 24 — 28. Morfologia del corfo uinano, Milan, 1891, pp. 113 
 — 119. Commentari di Clinica Medica, Vol. I., Padua, 1888, pp. 36 — 62, 
 Vol. II., Padua, 1893, pp. 75—177. 
 
 t Jaccoud, Cours de Patholoqie, Le<;on du li Nov., 1882. Paris, 1893, p. 15. 
 
 J Jaccoud, Clinique medicale de la Pitii, pour 1883, 1884, Paris, 1885, 
 p., 23, et pour 1%%^ — 86, Paris, 1887, pp. 94 — 98. 
 
 § Jaccoud, Lejon du 5 Mai, i888 (Bulletin Midical, 11 Nov., 1891, p. 1034. 
 
194 BACTERIA OF THE SPUTA 
 
 originators of effects which are always identical; (3) with reference 
 to the supposed immobiUty of the species of bacteria. The 
 pathogenic microbes, he says, are not always extraneous to a 
 heahhy organism ; on the contrary, a large number of them nor- 
 mally lodge in it. The same disease, for instance, endocarditis, 
 may be induced by various microbes, and these, likewise, are not 
 unchangeable, but assume various forms. Thus, if certain patho- 
 genic microbes normally exist in the healthy organism, this fact 
 demonstrates that the diseases connected with them arise in us and 
 through us, else the disease would be a normal condition and the 
 healthy state a myth. The microbe, for Jaccoud, means only 
 the instrumental cause of the disease ; the how and not the why. 
 
 Nevertheless, micro-biology presents, according to the author, 
 quite a new character ; the transmissibility to others of a disease 
 originally autogenetic from some common injurious cause^a cold, 
 for instance, although even this fact is, in its turn, subject to the 
 law of a morbid spontaniety. * 
 
 Amongst the microbes which in a harmless state thrive in the 
 human organism, Jaccoud places the Pneurnococcus, Pneumobacillus, 
 Streptococci and Staphylococci, the Bacterium coli commune, and 
 numberless bacteria that swarm upon the skin and the mucous 
 membranes. He affirms that there is not a single one of these 
 microbes which may not become pathogenic in the affections of 
 the bowels or membranes, in pyaemia, or in erysipelas, rheuma- 
 tism, etc. The Bacterium coli commune, for instance, and its con- 
 geners may enter into the autogenesis of the intestinal affections, 
 causing catarrhal burning, and ulcerous affections, and even typhus 
 or cholera infections, cholera itself being not always characterised 
 by the presence of Bacillus virgula. 
 
 Then we are in danger of having a plurality of microbes 
 for the same disease, and a plurality of diseases for the same 
 microbe. This is the rock, says Jaccoud, against which the sup- 
 posed specific character of the microbic diseases is likely to be 
 wrecked. 
 
 The characters of a really specific disease (as poisonings 
 
 * Jaccoud, Note d, la Traduction de Graves, first edit., Paris, 1891, p. 142, 
 and Clinique Midicale de la Pitii, i886 — 87, Paris, 1888, pp. 52 — 55. 
 
AND CONTENTS OF THE MOUTH. 195 
 
 by animal venoms, eruptive fevers, syphilis, etc.) are, accord- 
 ing to the author, five :— Exteriority of the cause ; unity of 
 cause for each disease ; unity of effect for each cause ; immutable 
 relation between cause and effect; and constant reproduction 
 under the same species. Now, he concludes, it is manifest such 
 are not at all the characters of the microbic diseases ; whilst, on 
 the other hand, there is the fact that the greatest part of the 
 diseases truly specific {eruptive fevers, etc.) in any microbial cha- 
 racter is hitherto wanting.* Jaccoud means to indicate that the 
 diseases truly specific (eruptive fevers, etc.) present no peculiar 
 bacilli as their bacillar characteristic, whilst the contrary -should 
 take pjace, according to theory. 
 
 Middendorp's Remarks on Tubercle. — From the time that the 
 surprising researches of Prof. Koch upon the bacterial characters 
 of tuberculosis were made known, there have been opponents to 
 his theory. Prof. Spina, who was working with Prof. Strieker, 
 opposed it by observing that if the bacilli of Koch should be the 
 true cause of tuberculosis, they ought to precede the formation of 
 tubercular nodules. Let this precedence be disowned, and the 
 principal base of the new theory falls to the ground. Now, he 
 stated that he had never met with bacilli in the young tubercular 
 nodules, and that the bacilli found, by a further observation, in the 
 decayed tubercles and in sputa not only were mixed up with a mass 
 of other bacterial forms, but they did not even possess any truly 
 distinctive characters.t 
 
 Dr. Beale had already expressed a doubt respecting the exist- 
 ence of true giant cells. He considers those forms as heaps 
 resulting from the development and subdivision of nuclei (bio- 
 plasts) in 2i farmed material or stroma, slightly consistent and con- 
 tinuous in its mass ; and he adds, besides, that such cellules are 
 often wanting in many specimens, even typical of tubercles. J 
 
 * Jaccoud, De la Spontaneity dans les maladies microbiennes. Lepn 
 d'ouverture (Bulletin Medical, i6 Nov., 1892, pp. 1407 — 10), 
 
 t Spina, Studien ilber Tuberculosa, Wien, 1883. See also Wiener Med. 
 Presse, 1883, No. 19. Concerning the Experiments of the same author upon 
 the colouring of the bacilli in question, and the observations of Coppen Jones 
 and others, see later on. 
 
 t Beale, op. cit., pp. 330—31. 
 
196 BACTERIA OF THE SPUTA 
 
 Charrin pointed out that, from the early discovery of Koch, 
 numberless cases of acute miliary tuberculosis had taken place, in 
 which the characteristic bacilli within the tubercular nodules were 
 wanting; and this want was, from the beginning, attributed to 
 imperfection in the method of investigation ; that analogous facts 
 had happened later on to which the same explanation could not 
 apply. He afterwards quoted a case of acute miliary tuberculosis 
 in an adult, in which were found numerous tubercular granulations 
 without caseous degeneration or ulceration, and likewise without 
 the bacilli of Koch ; nor could such bacilli be discovered in any 
 way, even in the inoculated guinea pigs.* 
 
 Kuskow was also unable to find specific bacilli in the primitive 
 acute, miliary tuberculosis, when the granulations were isolated 
 within healthy tissues. He discovered them in cases of secondary 
 acute miliary tuberculosis, consequent upon a chronic tuberculosis. 
 He affirmed that the acute miliary tuberculosis does not at all 
 proceed from the bacilli of Koch, and that such bacilli only 
 appear there second-hand, proceeding from pre-existing tuber- 
 cular nodules.t 
 
 But the strongest opposition to the doctrine of Prof. Koch, on 
 the ground of pathological anatomy, is actually that of Prof. Mid- 
 dendorp, of Groningen. He has for several years maintained 
 that the bacilli of Koch are but a simple dissemination of buccal 
 microbes, as he had never detected them in the young tubercles, 
 which were not yet in communication with the bronchia. When 
 the lymph of Koch was so highly proclaimed, Middendorp thought 
 fit to divulge his researches ; but I shall not refer here to that keen 
 contest. Briefly, Middendorp, from the figures exhibited by Koch, 
 shows that the latter has found bacilli in tubercles only in a retro- 
 gressive phase, and already in communication with the air-passages, 
 and not in tubercles which were either partially or fully developed, 
 but still healthy and grey and isolated. The author then mentions 
 that he has never come across any such bacilli, either in young 
 tubercles, in yellow, caseous tubercles, or in cavities not communi- 
 cating with the bronchia. This would prove, according to him, 
 
 * Charrin, Comptes Rendus de la Sociiti de Biologic, 12 Oct., 1891. 
 t Kuskow, St. Petersburger Med. Wochenschr., 1891, No. 36. 
 
AND CONTENTS OF THE MOUTH. 197 
 
 that the relative bacterial germs, instead of preceding the evolu- 
 tion of the tubercle, are transplanted by second hand, and a 
 second time in the tubercular mass in decay from the nursery, 
 existing in the mouth ; and consequently they have nothing. to do 
 with the cause of the disease. Middendorp likewise impugned the 
 existence of true giant cells, for which he was rebuked, although 
 he was not the first to doubt the true or supposed origin of them.* 
 
 Middendorp treated largely upon the experiments of Villemin 
 and Waldenburg, about the inoculation of tuberculosis in animals, 
 adding reports of numerous inoculations, practised by himself 
 upon dogs. 
 
 To the censures made to him Middendorp replies a second 
 time, more particularly explaining the points relating to the origin 
 of the bacilli of Koch. " Here is (he says) the quintessence of 
 the problem concerning the doctrine of Koch on the bacillary 
 origin of tuberculosis, on the biological properties of these 
 microbes that he pretends to have discovered, and on the cura- 
 tive method founded by him. I deny the existence of bacilli in 
 sound tubercles of recent formation, yellow or caseous, as Koch 
 and the bacteriologists of his opinion would make us believe ; and 
 likewise I deny their presence in the same cavities, until these 
 enter in communication with the bronchia. Then the situation 
 changes ; but the bacilli found in such tubercular cavities, commu- 
 nicating with the bronchia, or in the sputa, are common bacilli of 
 buccal mucus, having, as such, reached the morbid seat, or in a state 
 of germs. In order to place our faith in the theory of Koch, that 
 such bacilli are in etiological relation with tuberculosis, he will have 
 to show them to us, not in the expectorated matter nor in the tuber- 
 cular mass taken from the walls of a cavity or cavern in communica- 
 tion with the bronchia, but in tubercles in a state of developme7it, or in 
 tubercles already fully developed, but still fresh, grey, raw, and 
 healthy. \ 
 
 * Middendorp, Le Remide de Koch, sa valeur, etc., Paris, 1891. Der 
 Werth des Koch 'schen Heilverfahrens gegen Tubercolose, Emden and Borkum, 
 1891. 
 
 t Middendorp, Nouvelks etudes concernant les bacilles tuberc, etc., Paris, 
 1 89 1. Weilere Mittheilungen, u. d. v. Prof. Koch Tuberkelbacillen, etc., 1891. 
 
198 BACTERIA OF THE SPUTA 
 
 In 1892-93, after the bitter criticism of the followers of Prof. 
 Koch, Middendorp wrote three pamphlets in Dutch, which were 
 translated and appeared in La France Midicale, in which he dealt 
 with the evolution of the tubercle, the giant cells, the so-called 
 pure cultures, and the lymph of Koch.* 
 
 Having again confirmed the absolute absence of specific bacilli 
 in tubercles still isolated, Middendorp enlarges, in rectifying some 
 inaccuracies in which he thinks Prof. Koch and his followers have 
 fallen, with regard to the structure and evolution of the tubercle. 
 Not even in the caseous and most inward part of the yellow 
 tubercles and their conglomerates did Middendorp detect bacilli ; 
 whilst an emulsion of 10 milligrammes of such caseous substance, 
 although deprived of bacilli, produced, when inoculated in dogs, 
 a miliary tuberculosis. He excludes a transmission by means of 
 invisible spores, not apt to colour, as well as the supposed transport 
 of tubercular bacilli (immobile) through migratory cellules. 
 
 In one of his letters to me, Prof. Middendorp thus expresses 
 himself : — " The bacilli only accompany pulmonary phthisis in 
 caverns communicating with the bronchia. They are, on one side, 
 immigrants just arrived there ; but for the greater part they are the 
 descendants of bacilli which first immigrated into these cavities 
 (which may on this account be compared to sinuous abnormal 
 recesses of the air-passages), and have produced there numerous 
 colonies of various generations. These microbes, therefore, cannot 
 be called tubercular bacilli, as they are not found in tubercles as 
 such, and have no etiological relation whatever with tuberculosis. 
 For this reason I maintain that tubercular bacilli, in the sense of 
 Prof. Robert Koch, do not exist at all." 
 
 With regard to cultures, Middendorp thinks that their virulence 
 is to be attributed, not to the bacilli in themselves, but to the 
 necrosing tubercular substance, in which the bacilli- are entangled. 
 Such substance, even free from bacilli, becomes extremely virulent. 
 It goes to form the cultures, and transplants itself in the succes- 
 
 * Middendorp, Tuberkelbacillen Bestaan Niet, Open Brief, etc., Gronin- 
 gen, 1892. Tuberkelbacillen Bestaan Niet, Vercfere Bij'iira^en, etc, , Gionineen, 
 1893. £s giebt Keine Wharen Tuberkelbacillen, etc., Groningen, 1894, La 
 cause (bacillaire ?) de la Tubet culose. Suite sur la doctrine erronie de R. Koch, 
 etc. ( France Midicale, 1S94, pp. 113 — 35). 
 
AND CONTENTS OF THE MOUTH. 199 
 
 sive transports, which cannot eliminate it or weaken its virulence. 
 It enters as well into the composition of the lymph proclaimed by 
 Prof. Koch for curing tuberculosis ; also,, its presence can be 
 proved in the cultures from which lymph is extracted. 
 
 Consequently (concludes the author) I cannot attach the least 
 value respecting the genesis of tuberculosis, to the inoculations 
 of these cultures, almost pure, because Professor Koch has not 
 inoculated the isolated parasite, but always conjointly with this 
 necrosing tubercular substance, specifically infectious. 
 
 So far Middendorp. But his definite conclusions leave, in my 
 humble opinion, two questions unsolved : the first concerning the 
 presence of peculiar bacilli resisting the decolourising agents in 
 sputa of consumption rather than in other sputa; the second ques- 
 tion, about their presence even in tubercular nodules of organs not 
 communicating with air-passages (glands, brain, etc.), impugned 
 by Middendorp, but proved by other accredited investigators. 
 
 These two points deserve the greatest attention. Even admit- 
 ting, with Prof. Middendorp, that such bacilli are not at all a 
 species distinct from other forms of Leptothrix (and I concur in 
 this opinion), it still remains to be explained why, in the tubercular 
 products, they especially resist the decolourising and are capable of 
 furnishing special cultures. Here is, in my opinion, the real point 
 of the question. I shall speak of this again later on.* 
 
 The Experiments of Pettenkofer and others on Asiatic Cholera. 
 
 The first who tried the ingestion of choleraic matter seems to 
 have been Rochefontaine, who swallowed pills of faecal matter of 
 cholera patients without any injury to himself. Similar experi- 
 ments were repeated by Klein at Bombay ; afterwards by Wall and 
 others. 
 
 During the epidemic at Hamburg in 1892, Prof. Pettenkofer 
 communicated to the Medical Society of Munich Emmerich's and 
 his own experiments with pure cultures of cholera bacilli in broth 
 taken from a quite fresh culture from Hamburg. Of this culture 
 
 * Lately the author has given a resume of his observations and views in a 
 newspaper : La Cause de la Tuherculose, Paris, Oct., 1895. 
 
200 BACTERIA OF THE SPUTA 
 
 Pettenkofer took a cubic centimeter in a solution of bicarbonate 
 of sodium, and suffered only from a slight diarrhoea, with many 
 comma bacilli in the dejecta. Emmerich repeated the experiment 
 in a dietetic excess, and exposed himself to a current of cold air 
 in the night. He suffered from a more acute diarrhoea, with bacilli 
 in the dejecta, and other disorders. 
 
 After these experiments, Pettenkofer dared not impugn, abso. 
 lutely, the pathogenic action of the Comma Bacillus. In a 
 letter to me, he maintains that the key of the etiological pro- 
 blem in the infectious diseases is to be looked for in bacteriology. 
 Therefore, he held the presence of bacilli in the dejecta to be a 
 proof of a certain connection with the choleraic process ; but he 
 did not acknowledge them as the only cause nor as the principal 
 seat of the virus. An epidemic, according to Pettenkofer, is an 
 equation of three incognitse : — x, the specific germ ; y, the local 
 conditions ; z, the individual predisposition. He attributed chiefly 
 to the want of these extrinsic conditions {y and z) the relative 
 harmlessness of the ingathered bacilli, Munich being at that time 
 free from epidemics. In order to confirm that interpretation, it 
 would have been necessary to repeat the experiment in infected 
 countries. Pettenkofer, referring to the experiments of Bouchard,* 
 concluded that the specific virus of cholera does not arise from the 
 Comma Bacillus, but is evolved in the human organism ; and, once 
 developed, that virus, taken directly from the body of cholera 
 patients, even when deprived of bacilli, transmits cholera to the 
 inoculated animals. f 
 
 Frsenkel objected that the disordelrs from which the two experi- 
 menters of Munich suffered should have been at once recognised 
 as true attacks of cholera, however slight ; but Pettenkofer had 
 previously requested Bauer and Ziemmsen to verify the nosogra- 
 
 * Bouchard, when inoculating substances secreted from the intestines or 
 kidneys of cholera patients, had noticed in the animals under experiment 
 symptoms of cholera ; whilst inoculating on other animals of the same species, 
 simple cultures of the Comma Bacillus, or the product of their material 
 change, had only obtained negative results. (Bouchard, Zes Microbes Patho- 
 gines, pp. 119—30). 
 
 t Pettenkofer, Ueher Cholera mit BerucksichtigHn^ d. jungslen Cholerapi- 
 demie in Hamburg (Munch. Med. Wochenschr., 1892, No. 46). 
 
AND CONTENTS OF THE MOUTH. 201 
 
 phic table of the indisposition from which they suffered. Emme- 
 rich added that the dejecta of the two experimenters, which con- 
 tained a great number of the comma Bacillus, had been poured in 
 a common closet without any disinfectants, and nevertheless 
 cholera had not broken out at Munich.* 
 
 These experiments were, in 1893, followed by those of Strieker 
 at Vienna, who, together with his six pupils, swallowed with 
 impunity large quantities of comma Bacillus. These experiments 
 were, however, impugned by Loeffler. 
 
 Lazarus, meanwhile, relates the case of an assistant who con- 
 tracted a slight cholera infection through having handled many 
 times choleraic cultures.f 
 
 Metschnikoff shortly afterwards published new experiments on 
 the subject.! Some considerable quantity of a culture nine years 
 old, and harmless to animals, having been swallowed in large 
 quantities, an attack of cholera was produced, which was followed, 
 on the tenth day, by recovery. This experiment was opposed by 
 Drasche ; but experienced practitioners proved the case to be a 
 real attack of cholera.§ Metschnikoff, in a second experiment, 
 obtained analogous results from a drop of a recent culture diluted 
 in broth. However, in various other experiments, he could only 
 detect slight disorders. Then he found that even large quantities 
 0/ choleraic cultures may be swallowed by man with impunity, the 
 result of a certain relative impunity being due, in his opinion, to 
 the presence in the intestinal passages of other antagonistic 
 microbes. 
 
 In September, 1894, whilst Hamburg was free from cholera, 
 Oergel, an assistant at the Institute of Hygiene, in making experi- 
 ments on guinea pigs, fell a victim to cholera after the seventh day. {| 
 
 With this exception, all the other infectious cases which hap- 
 pened in laboratories appear to have ended in the recovery of the 
 patient, and did not transmit the disease to other subjects. We 
 
 *Fr3enkel, Deutsche Med. Wochenschr., 1892, No. 48, Emmerich, ibid.. No. 50. 
 
 t Lazarus, Berliner Klin. Wochenschr., 1893, No. 51. 
 
 X Metschnikoff, Annates de Vtnstitut Pasteur, May and July, 1893 ; May 
 
 and August, 1894. 
 
 I Drasche, Wiener Med. Wochenschr., 1894. 
 
 II Deutsche Med. Wochenschr., 1S94, No. 41. 
 
 Q 
 
202 BACTERIA OF THE SPUTA 
 
 notice also that other observers detected comma bacilli identical 
 with those of cholera in persons who were simply affected by cos- 
 tiveness ; that De Crecchio at Naples found them in a case of 
 arsenical poisoning; and others (amongst whom Cunningham) 
 detected, in the dejecta of cholera, ten different bacterial charac- 
 teristics ; whilst, on the other hand, he met with cases of cholera 
 free from any traces of Spirilla or Comma Bacillus. 
 
 As we see, the facts proving the harmlessness, however acci- 
 dental and relative this may be, of the cultures and choleraic 
 matter, are neither few nor isolated, and we cannot pass them 
 over, particularly in a matter of such importance. It is difficult to 
 understand how choleraic cultures, ingested in large quantities, 
 may be harmless, whilst a few germs or bacilli introduced, whether 
 mixed with food, drinks, or inhaled, succeed, according to theory, 
 in producing the ravages of cholera. 
 
 The question being still sub judice, the mind naturally returns 
 to the opinion of Lewis (see the second memoir), who identified 
 the cholera bacilli with the normal comma bacilli of the mouth. 
 It was objected that these cannot be reproduced on artificial soils 
 of culture, whilst the cholera multiply there easily. But we can 
 meet this objection, in my opinion, by saying that these two forms 
 of bacilli represent a single bacterial element, though in two dis- 
 tinct phases of existence. In the original phase (buccal comma 
 bacillus) it is a question oi fertilising elements in full activity, 
 and therefore incapable of multiplying by themselves, though this 
 cannot prevent the same elements, when transplanted into the 
 intestine and afterwards on the cultural media, and precluded from 
 performing their copulative function, from falling back into a retro- 
 gressive phase of existence, reverting to simple vegetative elements, 
 and so multiplying by fission. In fact, some thought that the 
 cholera bacilli had an affinity to the Spirilla and Spirochcete, through 
 their aptitude to form articulated filaments, with short bendings.* 
 
 For other arguments against the bacterial origin of cholera I 
 refer the reader to the work of Wall.f 
 
 * See on this subject the forms of cholera spirilla from broth cultures, in 
 the drawings of Cornil and Babes (pp. cit. , third edition ). 
 
 t Wall, Asiatic Cholera : Its History, Pathology, and Modern Treatment. 
 Third edition, London, 1893. 
 
AND CONTENTS OF THE MOUTH. 203 
 
 Hypothesis of the Author on the Action of the Bacteria 
 upon the Human and Animal Economy. 
 
 Let us examine now whether an eventual simplification of the 
 bacterial' species, in the sense that I propose, may be made to agree 
 with laboratory experiments considered within their -proper and 
 strict confines. 
 
 It is not here the place to take into consideration the action of 
 bacteria in the external world and their destiny in the general plan 
 of nature ; as the disintegration of rocks, the elaboration of the 
 humus, the nitrification and other processes related to vegetal 
 chemistry, etc. Although we have such an array of facts to prove 
 .the great importance of Bacteriological Flora, its high destiny in 
 the universal economy, quite different from the supposed morbific 
 action, my task now is simply to consider bacteria in relation to 
 human and animal economy, under three aspects — viz., the healthy 
 state, the morbid state, and in laboratory experiments. 
 
 In the Healthy State.— \}ni^ex this first aspect I have elsewhere 
 noticed that the presence of bacteria in such a prodigious number, 
 especially in the mouth and the external genitals, exhibits the cha- 
 racter of a real excluding vegetation, destined to protect those 
 natural openings, and to spread itself into other and more distant 
 
 organs. 
 
 The presence of bacteria or their germs in the tissues and 
 fluids of the body had already been an argument for the considera- 
 tion and researches of many investigators. From a Memoir on 
 Relapsing Fever I quoted some passages from Dr. Beale, which 
 are worth repeating :— " The presence of these living organisms in 
 the tissues and fluids of the body is not peculiar to contagious 
 diseases. Bacteria and their germs have been found in numbers, 
 in cases characterised neither by fever, nor by specific symptoms, 
 nor by blood-poisoning, nor by being propagated by contagion. 
 Nay bacteria grow and multiply, and in all organisms and in all 
 parts of organisms, so very soon after death, either of a part or of 
 the whole body has occurred, as to render it almost certam that 
 their germs must have existed in the organism before death. They 
 live and multiply under many different conditions, and consume 
 healthy as well as morbid matters in a state of decay. In the 
 
204 BACTERIA OF THE SPUTA 
 
 internal parts of the bodies of man and animals, in their tissues 
 and in their blood, these little germs exist and probably multiply, 
 but very slowly, their life-changes being in abeyance as long as the 
 bioplasm of the body, with its higher life, prevails ; but when this 
 succumbs, the bacterium germ bursts its bonds, and its descen- 
 dants, soon numbering millions of bacteria, assert their sway to 
 the detriment of the part. That bacteria are not in themselves 
 hurtful has been proved most conclusively. Many things we eat 
 contain them in countless multitudes. In the alimentary canal 
 of infants suffering a little derangement of the stomach, bacteria 
 are often present in vast numbers, and in some of these cases not 
 a particle of the contents could be found at any point between the 
 mouth and the anus that was not teeming with bacteria. Many of 
 the secretions may contain them without any perceptible injury to 
 health, while hosts of them are invariably present in the fluids, 
 and in and about the superficial cells of the mucous membrane of 
 the mouth of all persons, even in the most vigorous health." * 
 
 To-day the presence of bacteria in the interior of single histo- 
 logical elements is so generally admitted that some wish to argue 
 a kind of equivocal generation or derivation of bacteria from the 
 normal granulations of the cellular protoplasm.f So B^champ 
 thought that bacteria were derived from microzymas, or protoplas- 
 mic granules, which were apt to transform themselves into bacteria 
 as soon as the nutritive activity of the cellule became extirict or 
 suspended. I An analogous hypothesis was advanced by Nuesch 
 — viz., that bacteria and ferments were only secondary cellules 
 produced by the normal cellules of the vegetal pulp, with this 
 difference, that bacteria would produce themselves when the albu- 
 menoids abound in the generating cellules (Zellsaftblaschen) ; and 
 when, on the other hand, glucose predominates, there would be 
 produced ferments. Sterilisation would only be an effect of 
 the extinction of the protoplasm of the generating cellule, without 
 
 * Beale, op. ctt., pp. 315—18. 
 
 tThe equivocal generation of the bacteria has, however, been refuted since 
 1880 by Dr. Beale ('see How to Work with the Microscope, London, 1880 
 p. 20s). 
 
 + Bechamp, Les Microzymas et les Zymases (Arch, de Physiologic, 1883). 
 
AND CONTENTS OF THE MOUTH. 205 
 
 the vitality of which the secondary cellules would be unable to 
 produce themselves.* 
 
 Even Prof, de Giovanni observed that, in the disintegration of 
 the red and colourless corpuscles of the blood, of the epitheliaand 
 cancerous cellules, they resolved themselves into other and more 
 minute elements, which would constitute \\\t plastidules of Haeckel, 
 the colonies of the bioplasts of Altmann, or the bacterial germs.f 
 
 Galippe, in three notes (from 1887 to 1891) treated of the 
 presence of micro-organisms in the vegetal healthy tissues ; and in 
 a fourth note (1893) described a method of his own for the research 
 of bacteria in normal living tissues, of vegetal or animal origin, 
 and especially in the testicle, liver, and kidneys, t 
 
 The researches of Escherich, Bumm, Mercit, Cohn, Neumann, 
 and Hauigmann revealed the presence of bacteria in the milk of 
 women in perfect health, which occasioned various conjectures 
 about their origin.§ 
 
 In the third Memoir I recorded the hypothesis of Rud and 
 Arndt — viz., that the moving granules of the salivary corpuscles 
 are but the germs of Spirochete lodging there, in the first phase of 
 their life. In the already recorded Memoir on Relapsing Fever I 
 myself described certain groups or heaps of colourless blood cor- 
 puscles, from which, by means of the oscillations impressed on the 
 preparation in pressing the thumbs, new Spirilla were, now and 
 then, seen to escape. || 
 
 These facts (and others which, for brevity's sake, I omit) are a 
 convincing proof of the vast diffusion of bacteria or their germs 
 in our economy, as if their universal presence or ubiquity were 
 co-ordinated to the various metamorphoses and operations of the 
 
 * Nuesch, Arch, des Sciences Phys. et Natu., Genfeve, 1886, No. 10. 
 
 t De Giovanni, Remarks concerning the Inflammaiory Process (Gazzetta 
 Med. Veneta, anno xxiv. e xxv. ). Altmann, Die Elementarorganismen ii. 
 ikre Bezieungen zH d. Zellen, Leipzig, iSgo. 
 
 X Galippe, Recherches et Notes Originates, Deuxifime S6rie, 1891, pp. 71 — 
 73, Troisi^me S^rie, 1S93, pp. 37 — 44. 
 
 § Hauigmann, Zeitschr. f. Hyg. u. Inf. Krank., 1893. See also Modern 
 Medicine and Bacteriotogicat World, Oct., 1893, pp. 252 — 54. 
 
 II Vicentini, On a Case of Relapsing Fever and on the Discovery of Spirilla 
 in the Blood, p. 23, and Fig. 2, h. 
 
206 BACTEKIA OF THE SPUTA 
 
 organism. In the second Memoir I touched on the influence of 
 bacteria upon digestion. Now, however, I must appeal to the 
 general law of cellular biology. 
 
 Considering the assimilating activity of bacteria, on the one 
 hand, and, on the other, their reducing activity at the expense of 
 the histological elements in decay, I have often asked myself 
 whether this ubiquity of the bacterial elements is not, perhaps, 
 destined to perform some important service, as, for instance, the 
 expurgation of the fluids to act as scavengers in relation to 
 decayed particles or cells of the body. 
 
 As is admitted, the elementary parts or cellules of the single 
 tissues have, each one of them, a fixed cycle of evolution, in which 
 they begin, multiply, and pass away, and other generations take 
 their place. We must therefore presume that the products of 
 decomposition of these extinct particles or cellulary corpses with 
 their residue become the pabulum of bacteria, in the same way as 
 is daily observed in the epithelia, in the corpuscles of the blood, 
 of the pus, etc. In similar manner the bacteria and their germs, 
 scattered in our bodies, would act as necrophagous agents, cleansing 
 the tissues and fluids of the economy, ijot only of the eliminated 
 substances, but even of the residue of the defunct histological 
 elements, which otherwise would contaminate the body. 
 
 The physiological destination in the organism being thus 
 understood, we possess the teleological reason of the presence and 
 ubiquitary circulation of their particles and germs in the various 
 points and recesses of the economy, following the passages and 
 . gaps of the blood and the lymph, or migrating slowly from an his- 
 tological element in decay into another contiguous element. Such 
 particles or germs may be so slender that, even when magnified to 
 2,000 diameters, they will not be detected * ; and can, therefore, 
 cross and pass through the cellular bodies and tissues as through 
 porcelain filters ; and more so, for they can penetrate through the 
 intestinal villi and the chyliferous system. 
 
 The origin of such circulating particles may be assigned to 
 those internal granules of the common bacterial articulations 
 which are drawn in Fig. 2, «'; in Fig. 4, h ; and in Fig. 5, e,e',f; 
 
 * Beale, The Microscope in Medicine, edit, cit., p. 318. 
 
AND CONTENTS OF THE MOUTH. .207 
 
 as well as to the young gemmules of reserve enclosed in the fila- 
 ments, before fully developing into new bacteria. Thus we have, 
 so to say, a fixed capital of bacterial forms, in remarkable or full 
 development, upon the digestive, respiratory, and genito-urinary 
 mucous membranes ; and a circulating capital of more rudiment- 
 ary, slender forms inside the body. 
 
 In Disease. — The preceding remarks may apply equally well 
 to the pathological conditions ; nay, we may presume that, in 
 local and general affections, the activity of bacteria may become 
 comparatively more intense and agitated. In a vast number of 
 morbid processes we have a great multiplication of histological 
 elements (phlogistic exudations, pus, etc.), and, consequently, a 
 tumultuous succession of new cellular generations, which rapidly 
 retrograde and disappear as the preceding ones. To this prolifi- 
 cation'^and retrogression would ensue, therefore, a larger number 
 of elements and bacterial germs ; and likewise a more rapid and 
 agitated multiplication of the same in the diseased parts. 
 
 The simple fact that bacteria are present in the products or 
 morbid seats does not imply anything specific, so long as their 
 totally'extraneous origin, and their quite independent entity, be 
 satisfactorily proved. Losing sight of this truth, as simple as it is 
 essential, one risks falling into strange exaggerations, and is likely 
 to include the most varied complaints (even the ingrowing nail) 
 in the list'of bacteriological infections.'^ 
 
 In the fever complaints, without proportional local incentive, a 
 similar thing happens. If in these cases the local processes are 
 relatively unimportant, there is, otherwise, an increase not l^ss 
 tumultuous, but even more general, of the cellular metamorphoses, 
 in many provinces or in certain systems of the economy ; thence 
 the hypertermia, with a rapid thinning and loss of weight in the 
 body. 
 
 This is the reason why, in the acute or chronic disease (for 
 example, tuberculosis), which more severely attack certain systems, 
 the presence of bacteria in the affected parts and other seats is 
 more easily detected. " Chacun suit," says Charrin, ''que les mal- 
 
 * On the bacterial origin of the ingrowing nail, see Regnault, Association 
 fran^aise pour Vavancement des Sciences, Cahen, August 13, 1894. 
 
208 BACTERIA OF THE SPUTA 
 
 adies ginkraks ou locales peuvent faire que nos tissues cessent d'etre 
 privlss de germes." * 
 
 Now, in my opinion, from this virulent deviation of the primi- 
 tive activity of the bacterial elements, concurring and multiplying 
 themselves in the affected parts, new properties which bacteria 
 assume would be evolved, often irritating, sometimes polluting, 
 and finally showing themselves through the inoculation of the 
 morbid products or cultures obtained from them in the animals 
 experimented upon. In other words, through the changes brought 
 about, harmless bacteria or saprophytes would develop into mfec- 
 tious or pathogenic bacteria. These contaminating properties 
 then are to be attributed, not to anything original and intrinsic to 
 bacteria, but to an accidental deviation of their activity. And 
 observe that the virulence so acquired would, in its turn, remain 
 still inactive, without any hurtful effect whatever, at least in most 
 cases, if the will of the experimenter did not help to deviate, as 
 far as possible, the natural course of vital phenomena by taking 
 up these bacteria, so modified, in order intentionally to transport 
 them, together with the relative products of decomposition, con- 
 trarily to the ordinary course of nature, into the animals experi- 
 mented upon. 
 
 These views largely agree with those of Goldscheider, who 
 believes that bacteria, originally harmless, become infectious or 
 pathogenic through the simple deviation of the primitive biological 
 properties impressed upon them by certain substances {predisposing 
 substances). Having once got this deviation, it will be maintained 
 from generation to generation, as the author proves in the Strepto- 
 cocci and Staphylococci.'^ 
 
 The possible affinity of saprophytic bacteria with pathogenic 
 forms are admitted in a work by one of Prof Koch's disciples. 
 "The distinction" (so writes K. Fraenkel) "between pathogenic and 
 non-pathogenic bacteria is not so absolute as it at first appears. 
 We know that a fair number of micro-organisms which seem 
 harmless may, in given circumstances, become pathogenic ; and 
 we also know, on the other hand, that some pathogenic species 
 
 * Charrin, Compt. Rend, de la Societe de Biologic, II Nov., 1893. 
 t Goldscheider, Centralblatt f. klin. Med., 1893, No. 33. 
 
AND CONTENTS OF THE MOUTH. 209 
 
 may lose their pathogenic properties, and form a series of harmless 
 bacteria." And elsewhere, " Considering that all pathogenic bac- 
 teria in a period more or less distant were lacking their pathogenic 
 properties ; that even the parasitic micro-organisms had originally 
 to lead a saprophytic life ; that the pathogenic action depends on 
 a special adaptation to the nutritive conditions, we might consider 
 the loss of this property as a reversion to their old way of existence. 
 It is clear that the tendency of the species to lose the virulence is 
 different, and that some keep it up longer than others. But for 
 all, the virulence constitutes only an accessory, apt to increase or 
 decrease, according to circumstances. Therefore the differences 
 in the virulence cannot assist us in separating, the one from the 
 other, identical species of bacteria."* 
 
 I will, on this point, recall an important distinction between 
 virulence and pathogenic action. 
 
 When, in laboratory experiments, we speak oi pathogenic or 
 non-pathogenic action of bacteria, we mean that we consider an 
 exchange of ideas takes place only as we implicitly hold the 
 action of bacterial cultures in the laboratory to be identical to that 
 of similar bacteria, in the genesis of the spontaneous infectious 
 diseases, as exhibited in daily practice, though the truth of this is 
 not quite demonstrated, as the fundamental point of the question 
 has not yet been proved — namely, the supposed aggression of 
 bacteria from without, or by first hand (say, in the pulmonitic or 
 tubercular diseases) ; nor is it even proved, in the affected part, 
 their precedence on the local alteration, as it results from the obser- 
 vations of Spina and Middendorp. 
 
 On the contrary, to say virulent action is to put the question in 
 its true terms. Thus, we only affirm the fact of certain alterations 
 following upon the inoculation, keeping it distinct from the other 
 fact of the preceding evolution of a spontaneous disease (from the 
 products of which we obtained the culture), and without attribut. 
 ing to this earlier fact what really belongs to the later one, by 
 second-hand — viz., to the irritating or polluting action of the 
 inoculated material. Though mostly unknown, the causes may 
 
 * Karl Fraenkel, Manual of Bacteriology, Ital. translat, Turin, 
 1892, pp. 109 — 123. 
 
210 BACTERIA OF THE SPUTA 
 
 be numberless by which a bacterial culture can prove hurtful to 
 animals experimented upon, and often the contamination that 
 follows has nothing in common with the original disease. 
 
 We must keep in mind two actual facts in laboratory experi- 
 ments. The first is that the bacterium inoculated in the labora- 
 tory is not a bacterium proceeding from the external world (as is 
 thought to be the case in spontaneous infections), but one pur- 
 posely taken ; and from what source ? From a morbid source or 
 from eliminated matter (sputum, pus, fceces, etc.). Now, such a 
 product or material is, after all, but a confusion of various elimin- 
 ated matters, either in an incipient state of decomposition or quite 
 decayed, destinated to be eliminated by the affected organism, 
 and not inoculated to a sound organism ; a material polluted, by 
 the cellular decayed bodies, incongruous to life, and even dele- 
 terious if taken from the dead body, as is often the case in such 
 experiments. 
 
 The second condition of the fact is this : — That a bacterium 
 hardly ever becomes virulent when it is brought back to these 
 sources, for it is originally destined by Nature, let us say, in the 
 mouth ; but it may become so when it is introduced into seats not 
 intended-«.ft, under the skin, in cavities, or in the circulation, but by 
 Nature, viz. :— The inoculation, as I said elsewhere, introduces at 
 once a huge mass (milliards) of bacterial elements into the internal 
 organs, where Nature did not intend them to be. What wonder, 
 then, that in a great many cases local or general alterations should 
 result ? If, on the contrary, you deposit the bacterial culture in 
 the mouth (a habitat destined by Nature for most of these minute 
 beings), it will prove harmless, as is demonstrated by the experi- 
 ments of Pettenkofer and others in reference to the Cholera 
 bacillus and those of Stagnitta, who for several days ingested rabid 
 matter, without any harm to himself.* The same thing happens 
 jvith the poison of serpents. 
 
 In Laboratory Experiments. 
 But it will be objected that the bacteria inoculated in labora- 
 tories were, for a certain time and for different generations, nursed 
 in artificial culture media, where it is presumed they may have 
 
 * Stagnitta, Bulletlmo deU'Acc. Med. di Roma, 1893, P- SZ°- 
 
AND CONTENTS OF THE MOUTH. 211 
 
 deposited every vestige of those putrid and deleterious substances 
 in which they were previously immersed. And this is partly true. 
 Thus, through frequent transplantations, the virulence of those 
 bacteria may be reduced and even destroyed. Again, the virulence 
 is an acquired property in the affected spots, and may, upon other 
 culture media, diminish or vanish. 
 
 But it is understood that such an epuration, catharsis, or 
 complete reversion of bacteria to their primitive harmlessness, 
 cannot be immediate ; nay, it may perhaps fail altogether. On 
 the other hand, we cannot advocate a purification process of this 
 kind for one property (the acquired virulence) without admitting 
 it implicitly even for the other (the supposed original pathogenic 
 action) ; as we cannot reasonably expect that, in the artificial 
 cultures, the bacterium should divest itself of the polluting 
 properties acquired in the pathological material (which are a 
 fact) without divesting itself at all of that original, pathogenic 
 efBciency, which is a supposition. 
 
 In truth, he who considers the conditions of the artificial cul- 
 tures must implicitly admit that, if the form, the type, or the 
 supposed specific entity of the bacterium and its supposed patho- 
 genic action is maintained intact, in passing from the morbid seat 
 or from the eliminated matter into the gelatine or broth, this 
 necessarily implies that, in those artificial culture media, are 
 exactly reproduced and maintained the same conditions, bio-phy- 
 sical and bio-chemical„ which existed in the product or in the 
 morbid seat. One of two things must naturally occur, either the 
 bacterium has degenerated from its primitive condition, or it has 
 not. If it is always the same, it cannot have preserved in quality 
 and quantity the same properties, but by assimilating the same 
 principles and casting in the nutritive vehicle the same products of 
 decomposition which first segregated (toxin, toxi-albumen, etc.). 
 The conditions of the bacterium as such then are not substantially 
 changed in the culture ; nay, a certain quantity of the impure or 
 decomposed material is transplanted with it ; and the same pro- 
 ducts of decomposition must necessarily be repeated, which, in 
 their turn, concur to produce the pollution, if they do not consti- 
 tute the principal cause, in the animals experimented upon. If it 
 were not so, the nature of bacterium would not always be the 
 
212 BACTERIA OF THE SPUTA 
 
 same, or, at most, the bacterium would be thoroughly modified in 
 its biological effects. In successive transplantations an analogous 
 transmission takes place ; a certain part of the fermentable mass, 
 together with the relative ferment, is transmitted from one culture 
 to another, as, for instance, happens with leaven or yeast used in 
 baking, which is transported from one parcel of dough to another. 
 A confirmation of this view can be obtained from the same 
 bio-chemical properties of the ordinary culture media. Let 
 us consider what are broth, gelatine, agar, serum, etc. Are they 
 not organic substances taken from vegetables or animals, living or 
 dead, which were, in both cases, destined to decay and perish after 
 being separated from the living economy ? The sterilisation may 
 have transitorily suspended such decomposition, but this decompo- 
 sition must initiate or recommence, so soon as a fresh corrupted 
 matter, as, for instance, a diseased product, any refuse or cadaver- 
 ous product, reaches it. In other words, on account of the mixing 
 up or dissemination of the suspected material, the culture medium, 
 although before sterilised, soon becomes decomposed ; and from 
 that decomposition no other than virulent and deleterious sub- 
 stances can be produced, to which, more than to the bacterium 
 itself, are to be attributed at last the poisonous effects and the 
 pollution in the animals experimented upon. 
 
 I have used the word pollution with good reason, because, in 
 many cases, the disorder caused by inoculations in animals 
 experimented upon resembles a poisoning rather than a disease in 
 the true sense of this word. 
 
 We must in fact make a distinction between the diseases which 
 are certainly inoculable, such as cow-pox, syphilis, tuberculosis, etc., 
 from those which are supposed to be inoculable (whooping cough, 
 pneumonia, etc.). The first are transmissible through graft- 
 ing the relative virus, either directly by means of the morbid 
 product in substance, or indirectly through bacterial cultures con- 
 taining that virus. In these cases it is really the disease which is 
 transmitted, and not an infection of another nature. The bacteria 
 taken from this source, having been in contact with the specific 
 virus, may transmit it from the sick to the healthy, as clothes and 
 linen transmit certain contagions, without constituting, however, 
 the generating principle of the disease. 
 
AND CONTENTS OF THE MOUTH. 213 
 
 It IS not the same with cases of the second series. Here it is 
 not a question of a communicated disease strictly speaking, but of a 
 produced poisoning, more or less analogous to the septic or cadav- 
 erous infection. This poisoning might partly resemble the original 
 disease from the products of which we got the culture ; if, as often 
 happens, the bacteria, removed from a tissue or an organ (in the 
 patient), will tend to regain (in the mouse or guinea pig) the tissue 
 or the homologous organ. It does not surprise us to find in those 
 organs or tissues of the animal the same bacteria as it would not 
 surprise a farmer (says Zeissl) to see the corn spring up where he 
 sowed it.* 
 
 Whether the animal succumbs or survives the experiment, it 
 will be difficult, in my opinion, to establish a perfect identity 
 between this disorder stirred up, mostly tumultuous, and the 
 evolutionary process of a true illness identical with the original 
 disease in man. There are numberless examples of supposed 
 bacilli or pathogenic spores (Letzerich) to prove how often the 
 experimenters are, on that point, led into most serious errors. 
 We may affirm that there is not a single one of them who, with 
 his spores or bacteria, has not pretended to have reproduced the 
 original disease (say whooping-cough), in all its characters, con- 
 futing one another in turn ; so easy is it to exchange the symptoms 
 of an artificial inoculation with those of a disease sui generis, 
 — for example, Septicoemia with hydrophobia (see second Memoir). 
 
 In that Memoir we have seen that there is no comparison 
 between the fabulous number of bacterial elements which, at 
 once, introduce themselves in the animals experimented on, and 
 the few homologous elements which, in the spontaneous infections, 
 are supposed to introduce themselves, one by one, from the exter- 
 nal world, given and not conceded, that in those spontaneous 
 infections the corresponding bacteria be of external origin, and 
 not simple disseminations of normal bacteria of the economy, as 
 seems more probable to me. In fact, bacteria are most frequently 
 to be found, and therefore considered characteristic of the disease, 
 in the affections of the respiratory and digestive passages. In 
 the diseases really infectious or more distinctly contagious (namely, 
 
 * M. von Zeissl, Sul Diplococco di Neisser, Ital. trans., Naples, 
 1887, page 46. 
 
214 BACTERIA OF THE SPUTA 
 
 the eruptive fevers and others mentioned by Jaccoud), a, true 
 bacterial characteristic has not yet been found. 
 
 Here I should stop at the so-called pure cultures. Apart from 
 the consideration that we do not even now know whether the bac- 
 teria hitherto known are true and complete beings, or simple par- 
 ticles of more complex beings, the difficulties of the isolation of 
 single types, or supposed bacterial species (which confusedly and 
 in huge masses encumber most preparations), are such and so 
 many that a culture can be called pure only in name.* 
 
 I have no sufficient data to give a positive opinion upon the 
 specification of Bad. anthracis, but fear it is nothing but B. subti- 
 lis modified either originally or consecutively in the pustule itself. 
 
 For many years it has been thought that the successful treatment 
 of Lister should depend purely and simply on the exclusion of 
 bacterial elements suspended in the atmospheric dust, which was 
 reputed to be the only source of the traumatic infections. Now, 
 this opinion is losing ground, and already many surgeons have 
 abandoned the spray, to direct every attention to cleanliness of the 
 hands, of the instruments, and of the medicaments applied.t 
 
 Moreover, it is nearly impossible to exclude efficiently the 
 bacteria from the parts operated upon (the same applies to many 
 manipulations in the laboratory), as it has been remarked by Dr. 
 Beale; for, as Galippe says, "// n'y a point de barriires pour les 
 microbes. X 
 
 *"I know nothing more humiliating than to be exposed to difficulties 
 apparently infinitesimal, which always reappear the moment we think we have 
 overcome them. Thus, after knowing the difficulties that the selection 
 
 of bacteria present, I have become very sceptical about the purity of cultures. 
 The methods boasted of a,s safe can, in my opinion, give only a rela- 
 tive security." — Miguel, Etude sur la fermentation ammoniacale, etc. (Annates 
 de Micrographie, 1891, /. 277). 
 
 tSee on this subject Lawson Tait, The Pathology and Treatment of 
 Diseases of the Ovaries, 1883, p. 268, and British Med.Journ., 1882; Ban- 
 tock, Medico-Chirurgical Transactions, Vol. LXiv. ; Haegler, Beiirage zur 
 Klin. Chirurgie, Bd. 9, 1892 ; Fritsch. Bericht u. d. Gincekolog. Oper., 1893 ; 
 Doderlein, Deutsche Med. Wochenschrift , 1893, No. 21, etc. 
 
 X Galippe, loc. cit., troisieme slrie, p. 37. On this subject I recall the 
 remarks of the same author upon a fungus 'ui generis (Monilia Sputicola), 
 which he saw germinating in filtered and sterilised saliva, attributed by him to 
 germs pre-existing in the filtering pipe (Journ. de I'Anatomie et de la Physiolog., 
 1885, Planche xxvii.) 
 
AND CONTENTS OF THE MOUTH. 215 
 
 Bizzozero pointed out, also, the presence of bacteria in the 
 same parts, carefully covered with Listerian dressings.* 
 
 "The practice of Lister " (wrote Dr. Beale) "is doubtless excel- 
 lent, but the data upon which it is based may nevertheless be faulty. 
 It seems very probable that the favourable results of this method 
 of treatment are due, not to the destruction of the bacteria, but to 
 the direct influence of the carbolic acid upon the wound. Car- 
 bolic acid interferes with the rapid growth and multiplication of 
 bioplasm, which results after a time in the formation of pus-cor- 
 puscles." + 
 
 Briefly, the theory of the direct contagium has nowadays sup- 
 planted that of the infection through the air-medium. Rather 
 than to germs floating in the atmosphere, the attention is now 
 directed to the organic residues that may be suspended there, or 
 may settle upon and contaminate the instruments, the hands, and 
 the dressing materials (unclean particles, incrustations of pus, 
 exhalations, and excretions). Perhaps the anomalous bioplasts of 
 Dr. Beale may have something to do with it. 
 
 But even when the traumatic infections should only be due to 
 bacteria, the nature, essentially, and originally pathogenic of them 
 would not at all follow. Microbes totally inoffensive in their own 
 natural habitat may become offensive if, either by accident or pur- 
 posely, they are transplanted in one extraneous to it ; therefore, 
 harmless bacteria on the mucous membranes or on the skin, etc., 
 might, in the continuous solutions, in the serous cavities, or in the 
 blood, become corrupting because transplanted to locaUties neither 
 intended nor consented by Nature that they should occupy. 
 
 Concerning the modifications that common bacterial elements 
 may undergo in morbid parts {e.g., tuberculous), it is not difficult 
 to explain, seeing the special bio-chemical and bio-physical condi- 
 tions which on their arrival they find already established there. 
 The special resistance of tubercular baciUi to the decolourising 
 test and other analogous facts fall in, I think, under this heading. 
 I thought it probable that such baciUi might in part be derived 
 from the same sporules of Leptothrix (as they are very susceptible 
 
 * Bizzozero, Man. di Microscopia dinica, ist edition, p. 50. 
 t Beale, op. cit., p. 316. 
 
216 BACTERIA OF THE SPUTA 
 
 to aniline colours), fallen in the tubercular seats, and there mul- 
 tiplying in series, under special conditions at present unknown ; 
 and partly (those with cancellated appearance) should be derived 
 from other Leptothrical elements still not determined, and modi- 
 fied in an analogous manner. But it is yet to be explained in which 
 way both of them acquired, in the tubercular seats, a special apti- 
 tude for retaining the aniline colours.* However, admitting the 
 circulation of common bacterial elements in the internal economy, 
 the presence of the bacilli of Koch would be fully explained, even 
 in the tubercles of the brain, glands, etc., without having to admit 
 their specific origin or their etiological value. 
 
 In summing up the reasons hitherto expounded, I venture to 
 conclude — (i) That the pathogenic efficiency! of the so-called 
 pathogenic bacteria, in the genesis of the infectious spontaneous 
 diseases (as exhibited by the clinic), is not at all demonstrated ; 
 (2) That the laboratory experiments show, neverthele,ss, a very 
 important fact — namely, certain inquinating properties of some 
 bacterial cultures ; (3) that this order of facts, relative to the 
 artificial infections, this power of transmissibility of a morbid quid, 
 is worth the greatest attention, but cannot, by itself alone, be taken 
 as a distinctive criterion of the bacterial species. 
 
 * I refer here to the researches of Prof. Spina on the colourisation and 
 decolourisation of bacteria treated with tannin and other substances. " These 
 researches (he says) contradict the predominant rules in bacteriology. Hitherto 
 it has been said that the Schizomycetes resisting the action of nitric acid con- 
 stituted a definite species ; but the experiments just alluded to show, on the 
 contrary, that this distinctive criterium to characterise the micro-organisms has 
 no foundation, because the action of the same substance, the tannin, renders the 
 more different species resistant to acids." From other experiments not less 
 ingenious, the author concludes that bacteria may find even in altered tissues 
 (as, for instance, in the tubercular seats) such chemical conditions as will fix 
 the tint. Untersuch. u. d. Entfarbbarkeit d. m. Anilinf. ting. Bad. ( Allg. 
 Wr. mediz. Zeitung, 1887, Nos. 15 and 16). Bacteriolog. Vers. m. gefarh. 
 Nahrsubstanzen (Ceniralblatt f. Bakt.u. Parasitenk. , 1887, Bd. 2, Nos. 2 and 3). 
 
AND CONTENTS OF THE MOUTH. 217 
 
 Some further Researches on the Morphology of Leptothrix, 
 
 After the publication of the third Italian Memoir, I had occa- 
 sion to gather some other facts concerning the inferior phases of 
 Leptothrix, which will be expounded later oij. I shall simply 
 record now a few supplementary remarks upon the already des- 
 cribed superior phases. 
 
 Together with the most obvious fructifications, with a thin 
 stem, I have drawn, in my last Memoir, other forms of ears, less 
 frequent, with a club shaped stem, having tv^o zones of colourisa- 
 tion, etc. (Fig. 27). In these, I was unable to detect the pedun- 
 cles, sterigmata, or implanted filaments of the sporules in the 
 central stem visible on the other forms. But later on, by a fortui- 
 tous chance, I was able to discern distinctly, even in ears with a 
 (r/«^-shaped stem, analogous peduncles, somewhat longer. It 
 occurred in this way: — 
 
 At the end of 1892, I had to examine the sputum of a 
 youth affected with an extended basal pneumonia of the right 
 side, and in the mass of that sputum I discovered numerous ears. 
 I think it was on account of the gravity of his condition and the 
 dryness of the mouth, phenomena having already intervened of a 
 metastasis in the meninges, which brought the patient to a rg.pid end. 
 I kept this sputum for twenty-two days, when, in examining the 
 wet preparation, coloured with gentian violet, using a 1/9 " dry " 
 objective and a condenser of i -30 N. A., I came across one ear 
 with a club-shaped stem, drawn in Fig. 32, a, and magnified 820 
 diameters. 
 
 This figure has the characteristic, which I had noticed in other 
 sputa and in the patina dentaria, of being lightly pyriforpi in 
 appearance. The ^//a (glair) shows a vividly coloured contour; 
 the central stem is also club-shaped, as in Fig. 27 ; but the club, 
 instead of being cylindrical in shape and of a uniform thickness, 
 exhibits a somewhat conical shape, with its base turned upwards ; 
 thence its general pyriform appearance. I noticed that such ■ 
 conical club-shaped ears are, generally, longer than thpse with 
 cylindrical clubs. 
 
 As the colourisation had been too strong, and the preparation 
 was full of numberless stained granulations, I clarified it with a 
 
218 BACTERIA OF THE SPUTA 
 
 drop of liquor potassce through capillarity (i : lo), always keeping 
 the ear in the visual field. As soon as the solution of potash 
 reached that field, the coloured granules at once disappeared, and, 
 in spite of the small enlargement, I was agreeably surprised to 
 see, at the same time, the glia discoloured, and the thick and very 
 distinct peduncles from the two sides of the central stem, make 
 their appearance, as in Fig. 2,2, b. 
 
 In the first place, the dissolving action of potash only acted on 
 the glia, the peduncles and the sporules still keeping the stain ; 
 but afterwards they also became pale, as I ascertained by using a 
 i/2S-in. objective, although they were discernible, specially by 
 lamp-light. The central stem kept the stain longer, only fading 
 within four hours. 
 
 I have already remarked that, even in the Patina dentaria, ears 
 of the same form are to be found, and upon these as well as upon 
 those with the cylindrical club (Fig. 27) I verified the fact that 
 potash acted in the same way. . This re-agent showed me, besides, 
 that the da^-j with club-shaped stem in both varieties are, in the 
 dental patina, more frequent than I had thought. It prevents the 
 drying-up of the specimen, and thus allows one to study its par- 
 ticularities for several days through a residual sufl"usion of a light 
 azure, which is maintained in the central stem, in the peduncles, 
 and the spores. 
 
 After decolouration with potash — viz., after four or five hours — 
 we may obtain another satisfactory effect by letting penetrate, at 
 intervals, under the cover, three drops of the iodurated solution of 
 iodine, which, within another hour, reproduces again the violet 
 colour, though not to the same extent as at first. 
 
 With regard to the affinity of the spores in question for aniline 
 colours and their analogy on this point, with the bacilli of Koch, 
 I will give other examples. 
 
 I have said in the Second Memoir that, at another time, gly- 
 cerine may be substituted for water (in the preparations of sputum 
 as in those of the Patina dentaria), letting it penetrate through capil- 
 larity. I preserved some similar preparations for several months, 
 and was enabled to remark that the common bacteria and the 
 Pneumococci, not capsulated, are the first to decolourise (within two 
 or three weeks), under the action of glycerine; the large Diplo- 
 
AND CONTENTS OF THE MOUTH. 219 
 
 cocci (forms of Sarcina), coloured in a darker blue tint, hardly 
 decolourise at all. The sporules of ears resist longer and only 
 fade within a month or two. Now, the preparations with tubercu- 
 lar sputum, dry, coloured with carbolic fuchsine and treated with 
 nitric acid and alcohol, when they are kept in glycerine, exhibit, 
 likewise, an analogous decolourisation. The Bacilli of Koch gra- 
 dually fade, and in about a month disappear altogether. The 
 fading is even quicker in the preparations with gentian violet. 
 The bacilli assume there a pale appearance within ten days, and 
 they disappear in two or three weeks. My experiments with dry 
 preparations and deep stainings continued for twelve to twenty 
 hours, and it is to be presumed that the resistance of the bacilli of 
 Koch would result even less in contrary conditions. 
 
 Elsewhei'e, I said that water, when used very warm, if pro- 
 tracted for a few minutes longer, becomes a good decolourising 
 means for the bacilli in question ; but even at the ordinary tem- 
 perature the water exerts, in the long run, the same action, as it is 
 proved by the dry preparations, kept without balsam, and re- 
 examined now and then, by means of a drop of water, through 
 capillarity. The repeated application of those drops of water is 
 sufficient to see the bacilli become pale and finally disappear. 
 
 With regard to the power of resisting the decolourising effects, 
 I remember a case in a preparation of foecal matter, proceeding 
 from a person believed to be suffering with intestinal tuberculosis. 
 The preparation, stained, by dry process, with carbolic fuchsine, 
 and then decolourised with nitric acid and alcohol, did not exhibit 
 any of the bacilli of Koch ; but in the midst of the mass of other 
 bacterial forms, more or less decolourised, certain colonies or 
 zooglceic groups of minute oval, pointed cocci were singularly 
 conspicuous; they were still brightly coloured and girded by 
 slender capsular appearances. 
 
 Towards the end of the Second Memoir, I recalled (in a note) 
 the observations of Klein and others on the forms of ramification 
 to be found in certain cultures of tubercular bacilli, and on the 
 probable analogy of such bacilli with higher micro-organisms or 
 fungi. Since then various other observers have described new mor- 
 phological particulars respecting the cultures of tubercular bacilli, 
 especially in regard to the formation of gemmules (Knospen) of 
 
220 BACTERIA OF THE SPUTA 
 
 elements of reserve (Kemartige Reservestoffe), and of protoplasm- 
 granules in the interior of baicUi ; in regard to the disposition of 
 vacuoles and knots, analogous to that of filaments of many fungi ; 
 to the development of a kind of mycelium and even of vertical fila- 
 ments (hypha) ; and, lastly, upon certain analogies of the tuber- 
 cular bacillus with the Actinomyces. According to these researches, 
 the bacillus of the tuberculosis would not be a Schizomycete, but 
 a mycelium-like fungus ; and Belfanti is inclined to classify the 
 tetanus bacillus in the same category. These views clearly, though 
 indirectly, agree with mine in regard to an eventual simplification 
 of the bacterial species ; but, for the sake of brevity, I will refer 
 the reader to two articles of Coppen Jones, where previous 
 remarks on this argument are illustrated by new and interesting 
 researches.* 
 
 I can say little about staining the ears with other tints. Car- 
 bolic fuchsine, with a wet process, vividly attacks the spores, but 
 hardly stains the peduncles. If, a second time, we let a drop of 
 lactic acid penetrate the object, the clods and the ears themselves 
 nearly decolourise altogether ; but we get the advantage of using 
 the preparation for a longer time, as the lactic acid keeps it wet 
 for several days. Methyl blue gives a fine transparency to the 
 specimens of the Patina dentaria and renders visible a larger 
 number of ears. It attacks the internal stem to a greater extent, 
 but has very little effect upon the peduncles. By substituting gly- 
 cerine a second time, we obtain a beautiful effect on the ears, 
 having a wrapping-cap at the top (Fig. 30), as glycerine promptly 
 decolourises the wrapping-cap, though leaving for some time the 
 internal ear coloured. We may speak later on of the advantages 
 of the methyl blue, for the examination of the bundles or matrix 
 of Lept<ah,riot. Hsematoxylin is not suitable for these researches 
 on account of its frequent precipitations. 
 
 I would remind those who wish to verify the existence and the 
 details of structure of the fructifications m question that they must 
 
 * Coppen JoilEs, Ueber einen neuen, b^i Tuber,kulose hij,ufigen Fadenpih. 
 ( CentralblcUt f. Bakt. u. Parasitenk., Bd. xiii.^ Nos. 21 anji 22). The same 
 author, Ueber die Morphologie u. Syst. StellUng des Tuberkelpilzes, etc. (In 
 the same Journal, Bd. xvu.', 1895. Nos. i and 2.) 
 
AND CONTENTS OF THE MOUTH. 221 
 
 be carefill to scrape lightly the patina deniaria, avoiding persons 
 with clean teeth and directly after tnastication. 
 
 Likewise, such fructifications are found even in sputa, and much 
 more frequently than I at first thought, provided that we examine 
 the sputum without previous colourisation, which causes the loss of 
 a great many ears. Those who know them can soon detect them, 
 even in the natural or unstained state, under a simple i/gth dry 
 objective and a 1-30 N.A. condenser. We can discern, also, their 
 complex mulberry apipearance, especially looking for them along 
 the edges of the preparation, where, by their lightness, they are 
 impelled. But the particle of the sputum must be so small as not 
 to pass over the edge of the cover-glass. The ears remain intact 
 a long time in the pulmonitic sputum. In one case I detected 
 them on the sixth d&yi and, in the sputum of the above-mentioned 
 youth, on the twenty-second day after emission. But numerous 
 ears can be found even in sputa of bronchitis and in the same 
 tubercular sputa examined in the natural state, and by the wet 
 process. Some were found even in emulsion of sputa with soda, 
 or potash during my research for elastic pulmonary fibres, and 
 lately I think I have found also some specimens in faeces. • 
 
 At a first glance, our ears might be taken for certain rarely seen 
 specimens, which somewhat resemble them, as Fig. 33 shows 
 (deeply stained with gentian violet, magnified to 820 diameters). 
 Here it is simply a question of two rows~ of cocci in mechanical 
 superpositibn to certain large filaments or bacilli, of opaque 
 apipeatance and probably old, with some other cocci here and 
 there on the central line, with no trace of proper stem or involving 
 glia. These incidental aggregations of more considerable dimen- 
 sions (which we might caW false ears) have nothing, apparently, in 
 common with true fructifications. 
 
 In diabetic and albuminous urines, I had already found, in 
 many instances, some small and isolated zoogloeic groups or 
 branches, formed by two or three longitudinal series of bacteria 
 and bacilli, coarsely resembling our Spica or ears, as described in 
 the patina dentaria. But lately I met, by chance, with a con 
 spicuous and beautiful growth of this kind (resembling a true 
 zodglma ramigera), in a diabetic urine, with many crystals of uric 
 acid, albumen, and some red blood-corpuscles. This growth, with 
 
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238 INDEX. 
 
 PAGE 
 
 Ramification in Tubercle Bacilli .. ••• •■• 141 and 219 
 
 Ranvier on Myelin ... ... ... • ■•■ ^3 
 
 Recapitulation of Arguments in the Second Memoir ... ■•• '37 
 
 Recognition of the Source of the Sputa, Necessity for ... ... 9° 
 
 Researches, New, On the Fructification of the Normal Parasite of 
 
 the Mouth ... ... ... ... ■•. ■•• '66 
 
 Researches on Expired Air ... ... ... •.• ••■ 35 
 
 Researches on the Morphology of Leptothrix, Further ... ... 217 
 
 Reasons for Writing the Appendix ... ... ... ... 187 
 
 Roots and Haustoria, Forms of, in some Filaments of Leptothrix ... 116 
 
 Saliva, Collecting and Preparation of ... ... ... ... 95 
 
 Secondary Micro-Organisms of the Mouth ... ... ... lo6 
 
 Sheath Expansion or Young Antheridium ... ... ... 176 
 
 Spermatic Fluid, Capsulated Diplococci in ... .. ... $1 
 
 Spina on Tubercle Bacilli ... ... ... ... 195 and 216 
 
 Spirillum sputigenum ... ... ... ... ... 103 
 
 Spirochaete dentium ... ... ... ... ... ... 105 
 
 Spirochsete in Salivary Corpuscles ... ... ... ... 154 
 
 . Spirochete in White Blood-Corpuscles ... ... ... 205 
 
 Spores, Fructification by ... .. ... ... ... 124 
 
 S]5ores, Temporary and Persistent, Fructification of Leptothrix race- 
 
 mosa by ... ... ... ... ... ... 172 
 
 Sputa, Collecting and Preparing ... . . 68, 90, 94, 169 
 
 Sputa from Pleurisy, Bacteria and Bacilli found in ... ... 52 
 
 Sputa, Forms of Gonococci in ... ... .. 20, 4I 
 
 Sputa, Further Remarks on the Bacteria and Bacilli found in ... 75 
 
 Sputa, Healthy, Bacteria and Bacilli found in ... ... 44 
 
 Sputa in Bronchitis, Bacteria and Bacilli found in ... ... 48 
 
 Sputa in Phthisis, Bacteria and Bacilli found in ... ... 54 
 
 Sputa, Pneumonitic Bacteria and Bacilli found in ... ... 49 
 
 Sputa, The Necessity for Recognising the Source of the ... ... 90 
 
 Staining and Preserving Specimens, Hints on ... ... 68, 94, 169 
 
 Staphylococcus pyogenes .,. ... ... ... ... 112 
 
 Staphylococcus pyogenes albus and aureus ... . ... 112 
 
 Sternberg's Paper before the American Microscopical Society ... 44 
 
 Summary of Investigations and Methods of Working ... ... 85 
 
 Tongue, Cuticle of the ... ... ... ... ... gg 
 
 Traumatic Infection, Bacteria in ... ... ... ... 214 
 
 Tubercle, Middendorp's, Remarks on ... .. ... ... jqc 
 
INDEX. 239 
 
 PAGE 
 
 Urethral Mucus, The Fructification of Leptothrix in Small Flakes of 134 
 
 Vegetation of Leptothrix racemosa. Phase of .. ... ... 181 
 
 Vignal's Fungi ... ... ... ... .. ... 108 
 
 Virulence and Pathogenic Action, The Distinction between ... 209 
 
 Whooping-cough, Bacteria and Bacilli found in ... ... ... 37 
 
 Whooping-cough, Cases in which the Sputa have been Exa- ) ,- i„ -, 
 mined i^' '' '° ^" 
 
 Whooping-cough, Myelin in ... 
 
 Whooping-cough, Observations and Considerations on the Fungi of.. 
 
 Whooping-cough, Previous Researches on the Sputa of 
 
 Whooping-cough, The Importance of Myelin in 
 
 Whooping-cough, The Sputa of 
 
 Wrapping-Cap on some Ears ... ... ... ... 179) 220 
 
 25 
 
 62 
 2 
 
 29 
 I 
 
 3finis. 
 
, BATH : 0. SEEBS AND SON, PRINTERS, 2 ARGTLB 
 
1 M 
 
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