Bacteriology

BACTERIOLOGY. Whatis a poisonous quantity ?--Possibly from about five to ten grains. Canyou fix the amount of sutph~te ~' cq t.er tlmt might be used with impunity ?--No, I never trief. Are there people peculiarly susceptible to pol runs ?--Ye'. inn sense. Persons suffering from stomach disorders might be more susceptible than others. Do you regard sulphate of copper as a cumulalive poison ? Witness : What do you call a cumulative poison ? Suppose you take sulphate of copper mixed with same food in small doses, and continued for several days~ would you naturally look for evil results ?--No. I presume you know Professor Simpson, of Glasgow University ?--I have that honour. Well, if I tell you that Professor Simpson said that the continued use of these peas from day to day would lead him to expect poisoning by copper, what would you say to that ? I should only say I would not expect it. Would you say that the whole of this compound passe~ through the bowels, and~nonewhatever goes into the system ? ~ N o ; you could never make a statement of that kind. Take this tin of peas ; suppose that the bulk of these peas, with their small proportion of copper which goes to the liver, suppose these are taken 8gain and a~ain, would that not show it to be cumulative ?--No, I think not, because it would pass from the liver again. Without injury to the liver ?--I think so. I suppose you have heard of people being poisoned by eating vegetables made in copper vessels ?~They used to be heard of formerly, but not recently. Can you tell me the reason why ?--How could I tell you the reason ? Suppose I suggest a reason.. Is it became iron vessels or vessels coated with tin are used now ?--I do not know. It does not frequently occur. The form in which copper is combined also materially influences the m~ximam dose capable of being taken with impunity. I consider the com. binatioa in peas the least active. Do you draw a distinction between the albumiaate in pickles or fruit and the albuminate in peas ?--There is no resemblance. They have nothing to do with each other at all. There is almost no albumen in such vegetables as generally occur in pickles, while there is a very large percentage in peas. There is almost none in fiuits such as apples, pears, etc. By Mr. Colquhoun : I have read Maestre's report, and I generally concur with the opinions therein. Dr. John D~ugaU, Professor of Materia Medica and Clinical Medicine in St. Mungo's College, and Examiner in Chemistry for the Faculty of Physicians, Glasgow, and Royal C01tege of Surgeons and Royal College of Physicians. Edinburgh, and Dr. John Glaister, Professor of Medical Jurisprudence and Lecturer on Public Health, St. Muogo's College, were examined, and gave corroborative evidence. Mr. Colquhoun intimated that he had also in attendance Dr. John Barlow, Professor of Physiology in St. Mungo's College ; Mr. Archibald Sloan, Dr. Angus M'Phee, Dr. Milne, and Dr. George M'Intyre, all of whom were held as concurring with the previous witness. This closed the evidence. T h e Sheriff, in giving judgment, said : I recently gave a detailed opinion upon this precise point, a n d I m a y not be entitled to refer to that case as in any way h e a r i n g on the present case, because each must be looked at according to its own evidence ; but that relieves m e from giving a n y detailed opinion to-day. I expected in this case that I would have had evidence on the two points obviously desiderated--flrst, ofpract:calinjury from eating these peas, and, second, of the difference between i ' 4 and 2'5. I have had n o n e , a n d the facts remain precisely as in that case. T h e opinions

i55

of the most e m i n e n t men, both in their writings a n d as given orally, are diverse to the utmost extent. I do not for a m o m e n t say that actual injury in every case must be necessarily proved to bring it within this Act, but where the opinions are so obviously diverse as here of the best authorities, it seems to, me imposdble to hold that it is proved that these peas have been r e n d e r e d injurious to health. I think the proof lies on Mr. Lindsay, and that, until he can prove thag, he c a n n o t ask the verdict which he craves in this case. I therefore assoilzie the respondent. I a m sorry I cannot give expenses, but I would have given them if I had h a d p o w e r . ~

Glasgow Herald: BACTERIOLOGY. ON CERTAIN MICROBIAN ASSOCIATIONS IN D I P h THERIA. By M. H. BARBIER (Arch. de M6decine Exp6rimentate). T . I I I . , No. 3. THE clinical forms which diphtheria m a y a s s u m e are, as is well known, numerous, a n d but little comparable among themselves. T h e state of the throat, the existence of secondary p h e n o m e n a - adenopathy, coryza, etc., present so m u c h differentiation, that o n e is tempted to see in each o f these types a new disease. Now our present knowledge of the biological a n d pathogenic properties of the Klebs-Loeffter bacillus allows to us the conception of a p u r e diphtheritic angina, having ever the same objective characters. Such is obtained experimentaIIy among animals, e.g., in the pigeon by inoculating on the previously w o u n d e d mucous m e m b r a n e of the pharynx, a pure diphtheritic culture, and such u n d e r certait~ circumstances is observed in man. But really more frequently we find other appearances, which ,would dismiss the idea of diphtheria were it not that certain clinical resemblances gave evidence of the action of the bacillus. Nevertheless, the opinion goes that the diphtheritic bacillus is not the only one taking part, a n d that other micro-organisms finding in the m u c o u s m e m b r a n e already affected, or in the exudations a favourable soil, in their turn enter o n the scene a n d give rise to contemporary accidents, without i m m e d i a t e connection with diphtheria. These complex cases represent the infectious forms of diphtheria, in contradistinction to the toxic forms of diphtheria, in which latter the bacillus diphtheria alone acts, a n d does so by its toxines. A m o n g micro-organisms found in the diphtheritic throat, some are found constantly during variable time, a n d then disappear. They do not correspond to a definite clinical type, and show themselves in even the most dissimilar cases. T h e y may therefore be considered as accidental parasites, for m a n y germs of the air and of t h e m o u t h may be m e t there ; others, on the contrary,

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B ACTERIOLOGY.

are only found in types of diphtheria identical in course and aspect ; their presence it seems influences the local lesion and the general phenomena. We have no longer to do with unimpqrtant microbes but true pathogenic agencies, for they are not only found on the mucous membranes, but in the secretions and in the tissues. Before the discovery of Klebs and Loeffler, and since their labours, all who have studied diphtheria have recognised the existence of micro-cocci in the false membranes, and about the connective and muscular tissues, in the lymphatic vessels, in the hlood, heart, liver, kidney, and in other organs blocking up the capillaries. We already know that the active erysipelas strep¢ococcus augments the virulence of the attenuated bacillus. Other microbes may act in an opposite way, and the question may be asked whether the bacillus of diphtheria does not give a temporary in$ectiousness to microbes of the tonsils and pharynx ~noffcnsive in their normal state. Our present study concerns three organisms which we have met with among others in variable frequency in diphtheria throats. These we wiU designate provisionaUy by the letters a, f3, y. Streptococcus a.--During May, I89o, the organism showed itself in nearIy all the cases examined by us, not only in the pronounced diphtheritics, but also in sore throats destitute of diphtheritic bacilli. In November of the same year it disappeared, to reappear in March, I89 L Hence it appears to be an accidental parasite. It develops in peptonised agar, and in gelatinised serum at 35 °, at the end of twenty-four hours the cultures being visible. The colonies on agar form translucid masses the size of grains of hemp seed, forming oblong prominences on weak cultures. They often resemble drops of starch paste. The development is slow at the laboratory temperature. At the end of a certain time the cultures become enfeebled, dry, and take the aspect of a "collerette" formed of a thin grey pellicle. In beef broth it forms flocculent deposit, without turbidity. It stains well by aniline and forms chains. The cocci have a tendency to elongate and take the appearance of bacilli. The size is larger than the following, and can be microscopically distinguished from it. Streptococcus f l ~ T h i s is found associated with a bacillus in diphtheritic throats when the mucous membrane is red, swollen, thickened with or without false membranes, the latter being often soft and diffluent. The tableau is completed by an adenopathy always very marked with infiltration of the cellular tissue. The serous exudation gives very rich cultures of the microbe. In three eases we have found it in the blood of the heart after death, and more of these on the vegetations of the mitral valve, the valve being affected by a re'_ent endo carditis.

On solid media the eolomes are very smallqthey consist of compressed heaps of transparent granulation% they are especially marked on agar, on wh;ch they often form a cloud. They develop in broth, forming deposits on the sides and bottom of the vessel without troubling sensibly the body of the liquid. Cultures lose rapidly thei r power of reproduction and their virulence. By successive stirring, the cultures at the end of a week remain sterile. Inoculation of cultures four clays old are innocuous. This organism is also met with in sore throats accompanied with white exudation, in which Loeffler's bacillus is absent. To ascertain its virulence, we have inoculated cultures of 24 to 48 hours' old into guinea-pigs in the cellular tissue of the thigh. All the animals died in three or four days, and died quicker than those inoculated with diphtheritic cultures derived from the same animal. The phenomena observed were as follows : From the second day an enormous swelling of the leg, with loss of movement and acute pain on pressure. The animals losing their vivacity remained crouching in their cages. The temperature at the commencement had a tendency to be below 380 , and at the approach of death it sank to 360 and 35 °, Sometimes there was at the last diarrhoea. At the autopsy, injection and infiltration of the leg ; along the psoas muscle, vivid vascularisation in the vicinity of the inoculated spot, swelling and redness of a ganglion situated on a level with the sacro-iliac articulation, and this inflammatory train reached in nearly every case to the hilum of the kidney. Only in one autopsy was there acommencement of suppurationqthe pus was liquid and bloody, nearly always local or generalised peritonitis, and often traces of pleurisy or of pericarditis. Liquid from the thigh, the peritoneal serum, and blood taken from the heart gave pure cultures of the streptococcus. The experiments were divided into three series :-(i) Guinea-pigs inoculated with the streptoCOCCUS.

(2) Guinea-pigs inoculated with diphtheria. (3) Guinea-pigs inoculated with diphtheria and the streptococcus. Culture Used for Inoculation.--Guinea-pig 3, from guinea-pig z and 2, dead 3. Guinea-pig 5, from guinea-pig 4 and 2, dead 2. Guinea-pig 8, from guinea-pig 7 and 6, dead 3. It is necessary to add that guinea-pig No. z was inoculated on the eleventh day of his diphtheritic inoculation with a culture of the streptococcus, and died in four days ; and the guinea pig No. 2 on the eleventh day of his streptococcus inoculation was inoculated with diphtheria and (by error) the streptococcus, and

BACTERIOLOGY. the animal died in four days. It hence appears that the streptococcus met with in diphtheria is endowed with virulent properties. Has this strcptococcus an action on the mucous membrane, and may it not favour the development of diphtheria ? If by means of a sterilised platinum cone a recent culture of the streptococcus is deposited on the mucous membrane of the vagina of the female guinea pig, a slight and curable vaginitis is devdoped, which manifests itself by redness and a muco-purulent discharge. If in the same way is applied a mixture of diphtheria and streptococcus, the next morning there is observed an intense vaginitis, which has the aspect of the pharyngeal inflammation in the serious form of diphtheria on the human subject. The irritation propagates itself to the vulva. The latter is covered with yellowish exudation, sometimes infiltrated with the colouring matter of the blood and of a red-brown colour. By means of a platinum needle, shreds of pseudomembrane may be detached. Death may supervene, or if the symptoms subside, the mucous membrane takes about the fifth or sixth day its normal hue, keeping only in places, at the entrance of the vagina ulcerations, covered with a greyish exudation which soon heal. We have also observed in one of the animals a fact which illustrates the part which the streptococcus may piny towards diphtheria. A female guinea-pig, which for four months previously had been used for the purpose of studying the action of the streptococcus on mucous membranes, and which had suffered from an experimental streptococcus vaginitis, received on the vagina a culture of diphtheritic bacillus. On the following day she pres.ented symptoms like those produced by a mixed inoculation, and died in convulsiom. This observation shows that the influence of the two infections need not be contemporaneous ; it puts in relief the importance which is to be attached to a past or latent streptococcus infection. The diphtheritic infection appears to give a new virulence to the streptococcus as a parasite on the mucous membrane. A simple sore throat may not only provide a favourable soil for the cultivation of the bacillus, but besides predisposes to the serious forms of the disease. Hence we may conceive three ways by which the mixed effect of this infection may be reaiized. (x) The bacillus instals itself on a mucous membrane modified by the streptococcus at an epoch more or less distant--the primitive infectiousform. (2) Or the bacillary infection is contemporaneous with the streptococcus infection m the

simultaneous inflcliaus form.

h (3) Or the streptococcus invades a diphtheritic t roat--thesecondary infectious [arm. Of these three forms the last is the best to take for clinical e~tamination.

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The cultures of streptococci in agar, those of diphtheria in serum, were mixed with a little sterilised broth, and inoculated in doses of { c.c. for the first two series, of a ~ c.c. for each of the two cultures of the third series, with the following result :-Di2hhtheria.-- 4 Guinea-pigs. Guinea-pig i, inoculated iuto the peritoneum,lived eleven days afterwards; guinea-pig 4, inoculated into the thigh, dead on the second day ; guinea-pig 7, inoculated into the thigh, died on the sixth day ; guinea-pig 2A, inoculated into the thigh, died on the third day. Slrep¢ococcus.--Guinea-pig 2, culture of six days old, living on eleventh day ; guinea-pig 6, culture of three days old, dead on the second day; guinea-pig xA, culture of three days old, dead on the third day. 2ficracoccus 7.--This is the most constant of the microbes associated with diphtheria. It may be obtained by culture of the false membranes, especially when the latter are accompanied by redness of the mucous membrane and muco-purulent excretion. They are met with also in sore throats with membrane in which the bacillus is absent, and of which we have already spoken of. Cultivated on serum,the colonies resemble a little those of diphtheria, roundish, isolated, of a yellowwhite but deepening with time, very apparent at the end ot twenty-four hours after exposure to 35°; by their confluence they form yellow manes. Their outlines are, however, not as defined as the diphtheritic cultures, and their development in a few days permits them to be distinguished from diphtheria. On agar they form at the end of twenty-four hours round colonies, thickest in the centre, yellow in colour, the colour deepening the following day. This microbe develops slowly in gelatin without liquefying it. Cultivated in broth, the broth becomes rapidly turbid, and on the bottom of the vessel a very abundant white deposit forms, whilst the bulk of the liquid retains its clearness. It is a coccus with a few short chains of three elements, or more rarely of five elemtnts. It does not appear, so far as the guinea-pig is concerned, to possess pathogenic properties. But it ~ffects mucous membranes, cauAng in guniea-pigs a mucopurulent vaginitis which lasts from five to six days, and then subsides. This vaginitis is an inlet for diphtheria, for in our experiments with the latter we have obtained false membranes. It is true there is no longer the intense vaginitis of the streptococcus. The mucous membrane is red, not hmmorrhagie, the thick vaginal discharge is moderated, and the false membranes are white and of a good aspect.

(2"0 be continued.)