On pseudo-tuberculosis, with special reference to pseudo-tuberculosis in birds

ON PSEUDO-TUBERCULOSIS, WITH SPECIAL REFER- ENCE TO PSEUDO-TUBERCULOSIS IN BIRDS.

By ROBERT MUIK, M.D., F.R.C.P.Ed., Lecturer o n Pathological Bacteyiology, and Senior Assistant to the Professor of Pathology, Edinburgh University.

(PLATES XVIII. AND XIX.)

THE term pseudo-tuberculosis is n o doubt scientifically inaccurate, but must a t present be employed for purposes of convenience. It is one which has been applied to a group of diseases in which the chief characteristic is the occurrence of nodules resembling tubercle more or less in their naked-eye appearance, but in which the causal agent is not the tubercle bacillus, nor the glanders bacillus, nor the organism of any of the well-recognised diseases. As a matter of fact, several conditions produced by entirely distinct organisms have been included under this head. There is, first of all, the disease described by Nocard, Parietti, Pfeiffer, and others, which is almost certainly the same as the tuberculose xooglo&pc of Malassez and Vignal, and it is perhaps in connection with this form that most work has been done. There is, secondly, that described by Cazal and Vaillard (’), the bacillus of which, obtained from the hunian subject, differs considerably from that of the previous disease, notably in that it liquefies gelatine media; the bacillus obtained by Legrain (z) from phthisical sputum is probably the same. Thirdly, there is a form of pseudo-tuberculosis described by Preisz (3), as occurring in the sheep, and another still by Kutscher-pseudo-tuberculosis muriunz (4) ; in each of these the organism is of different nature. Another bacillus obtained by Kutscher (5), from a glandered horse, and found by him to have pathogenic action resembling that of the glanders bacillus, may, from the lesions produced by it, be placed amongst the pseudo-tuberculosis bacilli. We niay also mention the form of pseudo-tuberculosis produced by an aspergillus as described by Kotljar (‘j) and others.

The organism which I have cultivated is almost certainly the same as that described by the writers on the first of these forms of pseudo- tuberculosis, and it is to this form especially that reference will be made. This form of pseudo-tuberculosis has chiefly been found as a natural disease occurring in guinea-pigs, rabbits, and other anirnals, or

PSE UDO-TUBERCULOSIS IN BIRDS. 161

has beeii produced accidentally by inoculation with material from a variety of sources, e.y. with garden earth, with sputum, with milk, etc. The bacillus which I have cultivated was obtained from an epidemic disease of birds, and it was only after making experiments on rodents that I became convinced of its identity with one of the bacilli of pseudo-tuberculosis. I n the present paper the subject-matter falls under two heads, the jifirst iiicIuding an account of the disease in its natural form, of the organism prodhcing it, and of its relations to the disease; th,e second, an account of the lesions produced experi- mentally in other animals, along with a consideration of the relation of the organism to others described. I n order to facilitate the discussioii of the subject, I give an abstract of the literature on pseudo-tuberculosis, practically coiifiiiing myself to those cases in which the bacillus described appears to be of the same nature as that under consideration.

Literature on pseudo-tuberculosis.-The first paper on this subject appears to be that of Malassez and Vignal (7, who, in a series of experiments, accidentally produced a disease by inoculation of a guinea-pig with a portion of a nodule taken from the arm of a tubercular child. I n this animal numerous tubercle-like nodules were found in the spleen, liver, omentum, and lymphatic glands, while in the lesions there were no tubercle bacilli, but small masses of somewhat homogeneous appearance, which they found to be zooglcea masses of short bacilli. Sometirncs these masses were found without cellular proliferation around. After a series of experiments they came to the conclusion that this was a distinct disease, and to it they give the name tuberculose zoogloLique. They isoIated the bacillus from the lesions, and produced the disease in its pure form. They described the organism as a short oval bacillus growing in chains, or in zooglcea masses. It does not stain by Gram’s method, and there is difficulty in demonstrating it in sections of the tissues, on account of the readiness with which it is tlecolorised.

Eberth (9) in 1885 described a corresponding condition occurring spon- taneously in a guinea-pig, and later (lo), a similar condition in a rabbit. He called the disease “ pseudo-tuberculosis,” and thought it probably the same as that described by Malassez and Vignal. He, however, made no cultures. Nocard (11) also, in examining the lungs of a fowl studded with numerous small nodules, found a microscopic appearance similar to that of the first- named writers, and considered the disease to be the same as that described by them. Nanfredi (12), 1886, from two cases of pneumonia obtained, along with Friedlander’s bacillus, an organism which he found to be highly pathogenic, producing tubercle-like lesions. He mentions that sparrows and quails are killed in from one to four days, without visible changes in the organs. His drawings and description correspond closely with those of Pfeiffer (see below).

Next, Chantemesse (13), 1887, produced a similar condition in guinea-pigs, by the intraperitoneal injection of the washings of some cotton-wool, through which air from a phthisical ward had been passed for some time. I n this case also, the nodules were present chiefly in the liver, spleen, and omentum, and he considered the disease to be the same as that of Malassez and Vignal. I n the following year Charrin and Roger (14) met with a similar condition in a guinea-pig which had died naturally. There were minute nodules in the liver and spleen, and corresponding changes were produced by inoculation with the bacillus which they cultivated. They found, however, that in the nodules formed, which they describe as foci of cellular proliferation without caseation, they could find no bacilli on microscopic examination of sections, though such

162 ROBERT MUIR.

could be seen in fresh scrapings. There were, certainly, no zoogloea masses. For this reason they supposed the disease to be different from that described by Malassez and Vignal. nor (15) in the same year described the disease as occurring in rabbits, and believed it to be the same as that of Charrin and Roger. He obtained cultures, but failed to reproduce the disease by inoculation with them.

Nocard and Masselin (IG) obtained the bacillus of tubereulose zooglobique from a calf which was supposed to be tubercular, but in which no nodules of any liind were found on post-mortem exaniination. The occurrence of this organism mas thus probably accidental. They found that on subcutaneous injection it did not kill pigeons, but did so on intravenous injection, zooglea masses being found in various organs. I n another note, Nocard (17) states that he has compared the organism of Charrin and Roger, of Dor, and of Granclier and Ledoux-Lbbard with his own bacillus, and found all of theni to be similar.

An important contribution was made to the subject in the same year by Grancher and I,edoux-L6bard (18). I n the course of certain experiments on the passage of tubercle bacilli through a layer of garden soil, tuberculose zoogZoLiyue was accidentally produced in a guinea-pig. They described the appearance of the cultures obtained, and especially the effect of intraperitoiieal injection in guinea-pigs. These effects correspond closely with what I have obtained, as will be described below. There is one point of difference, however, namely, that the resulting nodules sometimes showed zooglcea masses of the organism, sometimes only foci of cellular proliferation and degeneration, in which organisms were difficult to find. This latter condition was found especially after inoculating a number of guinea-pigs in series, the zooglcea masses gradually disappearing. Grancher and Ledoux-Lkbard agree with other writers in believing that thc psendo-tuberculosis of Charrin and Roger is really the same as tubemdose zooglo&igue.

Courniont (19) found a somewhat similar condition in the pleural membrane of a calf, and was able to reproduce the disease in guinea-pigs. He considered that the organism, though similar to, really differed from, that of Nalassez and Vignal. It is, however, not possible to decide from his description whether this is so or not. (Courmont’s bacillus is described separately by Kruse in Flugge’s ‘ Mikro-organismen ’ as the B. pseudo-tubewulosis siinzlis.)

A. Pfeiffer (20), in 1589, published a short monograph on pseudo-tuberculosis. He likewise produced the disease accidentally, on inoculating a guinea-pig with portions of tissue taken from a case of true glanders, the tissue having been removed from the body of the animal several days before. H e obtained cultures and reproduced the disease- in various animals, amongst which guinea-pigs and white mice were specially susceptible. The local lesion was a caseous swelling attended by a similar affection of the lymphatic glands, and the organs chiefly affected by the tubercle-like nodules were the spleen and liver. I n some instances he succeeded in producing the disease also by feeding with cultures, there being a lesion of the intestine, chiefly in its lymphoid tissue, a nodular affection of the peritoneum, etc. He makes no mention of the occurrence of the zoonlea masses, nor does he mention the possible relation to fuberculose z o o g l k p e . There can be little doubt, however, that his bacillus is the same as that of the French writers.

Zagari (”) describes a disease which he found as a natural affection in rabbits, and which he believed to be the same as that described by Malassez and Vignal. He mentions that he reproduced the disease in birds, but does not mention the species. (Pfeiffer failed to infect pigeons.) Parietti (z2),

by injecting a rabbit with milk in the course of cerhin experiments, produced lesions very similar to those described by Pfeiffer. Here, again, I think there can be no doubt that the two organisms are identical. Meguin and Mosny ( z 3 )

at the second Tuberculosis Conference mentioned the occurrence of a form of pseudo-tuberculosis in hares, and state that they were able to reproduce the


disease in guinea-pigs. A considerable amount of interest is attached to a case of pseudo-tuberculosis in the human subject described by Hayeni (2 ” , and clainied by him to be the first recorded. The patient was a young nian who suffered from symptoms of gastro-enteritis and persistent algidity, terininating fatally. Post-mortem the left suprarenal capsule was found to be enlarged and caseous, and Peyer’s patches and solitary glands were also affected. KO tubercle bacilli mere present, but another bacillus was found microscopically in the tissues, and obtained in pure cultures from the lesions, as well as from the blood. The organism was found to be similar to that of Malassez and Vignal, and in guinea-pigs produced effects similar to those described above. This observation makes it possible that cases in the huinan subject have hitherto been overlooked.

So far as one can judge from an abstract in the Centrallil. J Bakteriol. 71. Parasitenk., i t is chiefly concerned with a discussion of the different varieties of the disease without any original observations.

Preisz (26 ) , in a second paper on pseudo-tuberculosis, mentions the fact that he has obtained cultures of the bacillus of pseudo-tuberculosis of Nocard, of l’feiffer, of Zagari, and of Parietti, and on carefully comparing them has found them to be identical as regards their morphological characters, their appearances in cultures, and their pathogenic effects. Lastly, Delbanco, in a recent paper (27), describes the disease as occurring as an epidemic amongst guinea- pigs. He cultivated the bacillus, and studied its pathogenic effects, especially those produced by feeding animals with cultures. His bacillus is the same as Pfeiffer’s, and his general results closely correspond.

Possibly this also is of the same nature.

Pseudo-tuberculosis is the subject of a thesis by Plancard (?”.

Froin the observations of the various writers nientioned above, it will be seen that the disease described has been produced by a bacillus obtained in a great variety of ways. I n the tissues there are nodules which resemble tubercle nodules in their naked-eye appearance, though less in their microscopical structure. The bacillus grows readily on ordinary media, and does not liquefy gelatine. In guinea-pigs the lesions occur in two varieties, namely, one in which zooglea inasses of bacilli are present, and one in which there are foci of cellular proliferation with few bacilli. It has been found that the one form can pass into the other, but I am not aware of any observation showing on what this difference depends. A very characteristic affection is produced by the intraperitoneal injection in guinea-pigs, and in all cases the spleen and liver are the organs which have a special liability to be affected. The distribution of the organism in nature is not fully known, though it appears to be pretty wide-spread. The natural disease, as described by these writers, most frequently affects guinea-pigs and rabbits, though even in them it is by no means commou. I know of no detailed bacteriological account of the epidemic disease of birds t o be presently described, nor of any observations to show that the bacillus producing it is related to the bacillus of pseudo-tuberculosis of the above writers.

I. PSEUDO-TUBERCULOSIS IN BIRDS.

1. THE NATURAL DISEASE.-MY attention was first drawn to this condition by a medical student who brought to me a bird, the liver

164 ROBERT M U I . .

and spleen o l which contained numerous small yellowish nodules,- a condition which he said was common amongst singing-birds, and which was generally spoken of as tubercle. On staining sections of the organs I could find no trace of tubercle bacilli, the nodules appearing merely as finely granular masses without definite structure. Film preparations, however, froin the spleen of another bird similarly affected, when stained with a watery solution of gentian-violet, showed large numbers of short bacilli, and subsequently I found that the nodules were in great part formed by masses of them. Pure cultures of the organism were readily obtained.

Geneyal facts.-The disease is well known to bird-faiiciers as an extremely fatal form of epidemic, which occurs especially towards the end of summer and the beginning of autumn, though also at other times. I t affects a11 sorts of singing-birds, and sometimes nearly all the birds in a room may be killed off. The symptoms are said to be fever, general malaise, and those of intestinal irritation, the droppings being soft and of greenish colour. The affected birds sit with feathers puffed out, breathing hard ; they are apnthetic, refuse to take food, and generally die in from two t o five days after the first symptoms. The disease is variously known as “ bird plague,” “ septic fever,” ‘‘ tuberculosis,” etc. Dr. E. H. Clarke in a paper giving an account of the disease, and published in one of the poultry journals; mentions that he has found and demonstrated bacilli in the affected organs. He however made no cultures, and 1 can find no account of the subject in bacteriological literature. He mentions two forms of the disease, in one of which there are tubercle-like nodules in the organs, while in the other there are none present. I n birds said to have died of the second form, and sent to me for examination, I have failed to find any characteristic bacilli present. My observations accordingly apply only to those cases in which the organs contain the characteristic nodules.

Anatomical changes.-From an examination of between twenty and thirty birds dead of the disease, the following appear to be the main facts. The lesions are chiefly present in the spleen and the liver, the former being practically always affected in an extreme degree. The spleen, which in one of these birds is an exceedingly minute organ, is swollen to many times its natural size, reaching sometimes three-quarters of an inch in length and nearly a quarter of an inch in thickness. It is studded with numerous yellowish nodules, of which the largest are of the size of a large pin-head, whilst the smallest are just visible. The larger are the more distinctly yellow, and may project from the surface, giving the organ a. somewhat nodulated appearance. Usually marked congestion and sometimes haemorrhage are present around the nodules. The spleen is of fairly firm consistence, but distinctly friable ; the nodules, however, are tougher, and may retain their form when the organ is broken down.

“On Septic Fever in Birds,” A‘coltish Poultry Journal, July 19 and 26, 1895.


Sometimes only a few nodules are present, but generally the organ is crowded with them. The liver sometimes contains nodules in large numbers, but usually they are less iiumerous than in the spleen, and they are also of smaller size, being often just visible as small yellow points, which stand out distinctly on the dark surface. They are more iiuinerous in the superficial than in the deep parts of the organ. Often no nodules can be found in other organs, but occasionally the lungs contain a few,-small yellowish white points with an area of congestion around. Occasionally one or two are present also in the wall of the intestine, generally towards the lower part, being visible through the serous coat. At first I thought that this inight be a constant lesion and would explain the path of infection, but I have failed to find that it is so, having observed it only in four or five cases. In one case a nodule was present a t the lower end of the cesophagns, and one in the intestine just below the stomach. I have not found any nodules in any other part than those mentioned, but the detection of them in an animal whose organs are so sinall is a matter of considerable difficulty, and some may readily be overlooked.

Microscopic appearances.--In film preparations made from any of the nodules mentioned, and stained by any of the simple combina- tions of the basic aniline dyes, numerous bacilli are easily found, which have the following characters. They are of small size, being generally about 1.5 to 2 p in length, and about a quarter of that in thickness. Shorter forms are also present, some of which appear almost like cocci, and longer forms up to 5 p in length may be met with. They are seen lying singly, in pairs, sometimes in short chains, and often arranged in dense masses in which the individual organisms are with difficulty distinguished. They take up the basic aniline stains with great readiness, but lose the colour very rapidly when decolorising agents are employed. They are quite decolorised by Gram’s method. When stained somewhat faintly, the extremities often appear deeply coloured, the intervening portion remaining clear (Plate XVIII. Fig. 1). I may add that no tubercle bacilli can be found.

For examination of scetions, the tissues were fixed either in absolute alcohol or iii a saturated solution of corrosive sublimate, the latter being preferable. They were afterwards treated in the usual way and cut in paraffin.

The staining of the organisms in sections is somewhat difficult, as they so readily become decolorised. I have found the following to be the best method. A staining solution, 1 per cent. thionin-blue in 2 $ per cent. watery solution of carbolic acid, is made up; for use, this is dil&ed with an equal qiiantity of water, and filtered. Sections are stained for about ten minutes. They are then decolorised with weak acetic acid (a few drops in a bowl of water), mashed in water, dehydrated with aniline oil, passed through aniline oil and xylol (equal parts), then through xylol, and mounted in xylol balsam.

166 RUBER T MUIK.

I t is often unnecessary to decolorise at all. I may also mention the following method, which I have found applicable to all organisins which are easily decolorised. It depends upon the fact that if paraffin sections, sufficieritly thin, are floated on the surface of a watery staining solution, the stain penetrates the section and colours the various structures quite satisfactorily. The sections, after being cut and flattened in warm water, are placed on the surface of the staining solution for about half an hour. They are then washed well in water, placed on slides, and allowed to dry. The paraffin is then removed by xylol, and the section is mounted. Methylene or thionin- blue solutions are best suited for this method, as they do not readily overstain ; one or two trials with any stain are necessary to deteriiiine the length of time which gives the best results.

I n sections stained with ordinary tissue stains, say hzematoxylin with rubin and orange, the aodules, visible to the naked eye, appear as finely granular or almost homogeneous masses, staining of a dull purple colour with the combination mentioned, and showing no distinct structure. They might in fact be mistaken for masses of necrosed tissue. When stained with thionin-blue, etc., they are, however, seen to be neither more nor less than masses of bacilli, corresponding in character to those seen in the film preparations. They are densely packed together, except a t the margins, where the individual forms can be well made out. Short chains can sometimes be seen, but they do not occur to the same extent as figured by Grancher and Ledoux-L&barcl in tubercatlose xooglodipue. In the large masses, the centre is often stained very faintly, the organisms having died out there. (This would correspond to the zooglodes en couronne of French writers.) When separate masses have enlarged and partially coalesced to form a large area, the whole of the central portion may have necrosed, and in it portions of necrosed tissue may be seen between the masses. But no tissue can be seen in the interior of the individual masses. I n the spleen the condition appears to start with the growth of small irregular groups of bacilli in the pulp and sinuses, and these afterwards enlarge to form more or less rounded masses, often with irregularly sinuous margins (Plate XVIII. Fig. 3). A striking fact is that, beyond a slight amount of leucocyte accumulation and of congestion, with occasionally hzemorrhages, there is practically no change in the tissues around. In the liver, where the nodules are fewer, a similar condition is found. Small masses of bacilli settle in the hepatic capillaries a d in the smaller veins, and all stages can be seen, up to large rounded masses with pale centres and deeply stained periphery. Here again there appears to be little direct action on the tissues, the h e r cells lying in contact with the masses having an almost normal appearance. I n the lungs similar masses are occasionally found, and the same sequel of events can be traced, the growth of the bacilli starting in the capillaries. I n the kidneys often no bacilli can be found, but sometimes small masses can be seen, starting generally in the intertubular capillaries ; the masses, however, never reach a large size. The cells of the tubules may show cloudy swelling, with occasion-


ally granular disintegration, but these changes are not confined to the iieighbourhoocl of the bacilli. I have also once or twice found masses of bacilli forming rounded collections in the mucous and subniucous coat of the intestine, sometimes with necrosis of the superjacent epithelium, but I have never found deeply excavated ulcers, and the intestinal lesions always appeared to be of comparatively recent date, if one can judge from their extent.

The outstanding facts revealed by niicroscopic examination of the tissues affected are, accordingly, that the yellowish nodules visible to the naked eye are chiefly constituted of niasses of bacilli; that the distribution is by the blood vessels, and that there is remarkably little effect visible in the surrounding tissues. The histology of this disease in birds, therefore, differs very widely from that of tubercle.

2. CULTIVATION OF THE BACILLus.-This can be readily efiected by the ordinary gelatine plate cultures, or by successive strokes on sloped agar tubes. If inoculation be made from the organs in which there are visible lesions, nunierous colonies can be obtained. A few colonies can also be obtained from the blood of the heart after death, though usually the bacilli are there so scanty that their discovery by microscopic examination is a niatter of great difficulty. Cultures obtained from nearly a dozen birds, on being compared, were found to present the same characters, which may be described as follows :-

The bacillus grows well on most media a t the ordinary room temperature, though growth is much more rapid a t 37" to 38" C. Growth takes place both in a6robic and anwobic conditions, though i t is more vigorous in the former. I n gclntinr! stab cultures the growth is visible in twenty-four hours as a whitish line which increases in thickness, the deeper part consisting of a row of spherical yellowish- white colonies. Usually on the third day, a t 20" C. a delicate surface layer is seen spreading out around the puncture; this increases in extent so as t o form a round disc of transparent appearance, which inay reach the wall of the tube. The growth is closely similar to that of the typhoid bacillus, I n all the cultures the deep part of the growth becomes brownish yellow, and the superficial part more opaque ; occasionally feathery crystals are formed in the medium. Stroke cultures on sloped gelatine show a similar surface growth with somewhat sinuous margins. Shake cultures in gelatine give a general haziness in the medium, and there is no formation of gas.

I n gelatine plates the superficial ancl deep colonies are sharply distinguished. The former appear as thin transparent filnis with irregularly wavy margin, and may reach 2 mm. in diameter on the third day, after which they do not undergo much enlargement. Under a low power of the microscope the colony often shows an irregular darker point iii the centre, their margin is well defined, and their surface is slightly undulated and shining. Deep colonies, on the other hand, are spherical points of a slightly yellow colour, which, 011 the third day,

168 ROBERT MUIR.

reach scarcely 0.5 mm. in diameter ; this size is never much exceeded. Under a low power of the microscope, the deep colonies have a sharp circular margin of a brownish-yellow colour, and are slightly granular in appearance. Not infrequently there is a well-marked outer zone of somewhat homogeneous aspect surrounding the more granular central part from which it is sharply marked off, an appearance which recalls that of the ovum of a tape-worm.

A stroke culture on agny gives a whitish layer of growth, which is fairly thin and somewhat translucent ; the margin is somewhat undulated, and single colonies have little tendency to become fused. Growth takes place with practically equal readiness on ordinary, and 011 glycerine-agar. When an agar culture is broken down in water, it tends to form little fragments suspended in the fluid, and does not readily gire a uniform opalescence.

I n bouillon at 37" C. growth produces on the following day a general turbidity of the fluid, though the growth is more abundant below. Later, the broth becomes clearer, a pretty thick layer of somewhat flocculent deposit gathers a t the bottom, and a layer of growth adheres to the wall of the flask. Rarely, flocculent masses gather on the surface. The organism by its growth produces a slight degree of alkalinity ; thus a culture in neutral bouillon, after incuba- tion for a few days at 37" C., gives a distinctly alkaline reaction.

I n milk it grows readily without producing any visible change in the medium. I n lactose gelatiiie there is no fermentation or formation of gas.

On potatoes growt,h is always slow, and sonietimes does not take place at all. A large quantity of fresh culture must be transferred to the potato with the needle, in order to start the growth. The growth appears as a somewhat transparent layer, which becomes slightly yellowish in tint, It has slightly raised niargins, and does not spread far over the surface. I have never observed any appearance resembling that of the glanders bacillus, such as Preisz describes.

The micyoscopic appearance of. the organism varies somewhat in different media. I n hanging-drop cultures, after twenty-four hours a t 37" C., a great many of the bacilli are arranged in chains, whilst others are lying free. The latter show the usual oscillatory Brownian move- ment, but do not change their position in the field, and I have con- cluded, af ter careful examination, that the organism is non-motile. I have also failed to demonstrate the presence of flagella by any method.

In an agar culture the bacilli are in masses, and arranged irregularly (Plate XVIII. Fig. 2). They correspond pretty closely in character with those seen in the tissues, as described above, but they show more variation in size, and some may be of considerable length -short filaments of 6 p or more in length. A growth from potato shows pretty much the same characters-no chains being found. On gelatiiie the bacilli tend to be rather shorter.

PSE UDO-TUBER CUL OSIS IN BIRDS. 169

I have never found any evidence of spore formation. The organism may be exposed in a fluid medium to a temperature of 60" C. for an hour without being killed, but 70" C. for the same time is fatal.

3. EXPERIMENTS ON I<IRDS.-I have used for my experiments chiefly small birds, canaries, linnets, etc., and the experiments have been made both by subcutaneous inoculation, and by feeding with pure cultures of the organism described.

(a) Sdxxtaneous inoci~lntion.-Snbcutaiieous or, more correctly, superficial intramuscular (as it is very difficult to avoid injuring the muscle in small birds) injection of a drop or two of a suspension of a recent agar culture in sterilised water has always produced a fatal result in from two and a half to six days. The injections were made over the pectoral muscle. Symptoms of malaise appear generally on the second day, the bird sitting with feathers puffed out and respirations accelerated ; weakness and general lassitude increase, and the bird ceases to take food, and gradually sinks.

Post-mortem, there is great swelling with inflammatory induration a t the site of inoculation, the skin being stretched over the part, and more or less fixed. There is a yellowish, partly fluid, exudation under the skin, and, on cutting into the muscle, there is found to be extensive necrosis spreading from the surface downwards. The muscle fibres have a dull buff colonr and somewhat tough consistence, while the intramuscular septa are widened and stand out distinctly, showing a yellowish-white appearance, which is found on microscopic examination to be due to an inflammatory exudation along them, accompanied by the growth of enormous masses of bacilli. The inflammatory change may spread right down to the bone and even through the chest wall to the pleural membrane. If the inoculation has been made in the middle line, both pectorals are usually affected ; if it be made over one side only, the other is generally free a t the time of death.

Of five birds injected in this way, three showed nodules in the spleen, one of them also a few very minute points in the liver. Of these three, two died within three days after inoculation, and one on the fourth clay. Tn all three cases the cultures used were taken directly from diseased birds, and had not been passed through other animals. In one case the culture used was a recent one, in the other two the organism had been grown on gelatiiie for ten weeks, and showed no diminution of virulence.

The other two birds showed no nodules in the spleen visible to the naked eye, but in one of these small masses of bacilli were found in the spleen on microscopic examination. The two birds died on the fifth and sixth days respectively, and the cultures used had been passed through a white mouse. The virulence of the organism to birds had probably been modified in this way.

I n the cases where the spleen contained nodules, these had the same appearance as in the natural disease, but they were much fewer

1 TO ROBERT MUIR.

in number and of much smaller size, there being sometimes only one or two minute yellow points visible. The spleen was not so much enlarged as in the natural disease, was of dark purple colour, and somewhat soft. There are often small hmnorrhages in the lungs, and once I found a large diffuse hzemorrhage in the cranial bones. Xumerous colonies of the organism are obtained from the affected organs, and a few also from the blood of the heart.

Microscopic examination of films made from the juice of the affected muscle shows enormous numbers of bacilli, lencocytes, and red blood corpuscles, along with some broken-down connective tissue cells and portions of necrosed muscle-fibres. The bacilli are usually lying quite free, often in masses ; a few are contained within leucocytes ; but it may be mentioned that in the two cases where the organisms used had been passed through a white mouse, and apparently diminished in virulence, phagocytosis was very well marked. Sections of the affected muscle show an acute inflammatory condition affecting the subcutaneous tissue and extending along the intramuscular planes, caused by an extensive growth of the bacilli in the lymphatics. The muscle-fibres undergo necrosis with a sort of hyaline change, and afterwards break down. I n the intermuscular planes, the bacilli can be seen spreading out in large masses a t the sides ; while in the central part, both the tissue and the bacilli have undergone necrosis. The bacilli also invade the small veins, some of which are thrombosed and contain masses of them. The direct passage of the organism into the circulation can in this way be easily traced.

I n sections of the spleen, in the cases where nodules are visible to the naked eye, masses of bacilli are found as in the natural disease, though the masses are not so large. I n one of the cases in which no nodules were visible to the naked eye, small clumps of bacilli could be found adherent to the walls of the veiious sinuses. A few small masses may also be found in the liver, their growth starting in the hepatic capillaries. I n both organs the growth of the bacilli is attended by practically no tissue reaction.

There is, therefore, a wide difference in the action of the organism when in the lymphatics of the connective tissue, from that when it is within the blood vessels. A certain analogy might be drawn with conditions, such as anthrax in guinea-pigs, where there is local inflammatory reaction and a general septicamia ; only in this disease of birds the organisms, instead of being widely distributed in the general circulation, settle in the blood vessels of certain organs, and by their growth form large masses.

Summnry.-The organisms produce, locally, an intense spreading inflammation with extensive necrosis of muscle. They grow in large masses in the lymphatics of the connective tissue ; they gain entrance to the blood, and settle especially in the liver and in the spleen. They may produce small nodules, which may be seen as early as the


third clay after inoculation ; or death may take place, from the extent of the local lesion, without the typical anatomical changes being produced in the internal organs. All the parts affected, as well as the blood of the heart, give cultures of the same organism.

( b ) Feeding experinients.-I have made experiments by feeding the birds with the organism, both by breaking down little portions of the diseased organs in the drinking water, and by mixing with the water a recent culture of the organism on agar. A fatal result has always followed, generally on the tenth to the fourteenth day. Several experiments of this kind were made towards the end of summer, but, as the disease was not uncommon a t the time, there .was still the possibility that the birds might have become infected otherwise-a possibility which was practically excluded by a consideration of the regular times a t which the birds died. I waited, however, till winter, and obtained five healthy canaries from a bird-dealer, who stated that he had seen no traces of the disease for many weeks. Two of these were injected subcutaneously, and died within three days, as already described, with very early nodules in the spleen. The other three, put in a new cage, and kept for several days, were found to be quite healthy. A fresh agar culture of the organism was then broken down in their drinking water. One bird began to show symptoms on the sixth day after, being rather less lively and appearing thicker in the body, owing to its feathers being slightly puffed out. On the two following days these symptoms were more marked, the bird sitting immobile on its perch with its feathers erect. On the ninth day it had descended to the corner of the cage, and showed difficulty in respiration ; it died in the afternoon of the same day. A second bird showed the same symptoms a day later, i.e. on the seventh day, and died on the tenth. The remaining bird was ill on the tenth day, and died on the thirteenth. None of them showed any intestinal symptoms, the droppings being scarcely moister than usual, though such symptoms were present in some of the earlier experiments performed in summer. The symptoms produced by feeding with cultures correspond in nature and duration with those of the natural disease.

It is unnecessary to give in detail the post-mortem appearances, as they corresponded in every respect with those described above as occurring in the natural disease. I n all cases the spleen was enormously enlarged, and studded with yellow noclules of various sizes. The liver in all three showed a few scattered yellow points of minute size. No other nodules could be found on naked-eye examination. The intestines were distended with fluid contents, and in one case there was an admixture of blood, but no nodules could be seen in their wall, nor could any collections of bacilli be found on microscopic examination, though a few might have been present and escaped notice. Accordingly, in the disease produced by feeding with the bacilli as well as in the natural disease, I cannot state definitely

172 ROBERT MUIR.

whether there is first of all a distinct intestinal lesion, or whether the organism gains entrance to the circulation without multiplying to any extent in the tissues of the intestinal wall. The nodules in the spleens and livers showed masses of bacilli as usual ; these had the usual characters. Cultures of the bacillus were obtained from the affected parts.

If we consider the changes found in these five birds, as described, we cannot but conclude that all the animals were free from the disease at the time of the experiments, a8 the symptoms and anatomical changes completely harmonise with the time and method of inoculation. .

GEPI'ERAL suMManY.-This disease of birds, which I have called " pseudo-tuberculosis of birds," and which is characterised especially by the presence of yellowish tubercle-like nodules in the liver and spleen, is caused by the bacillus which I have cultivated, and of which the characters are given above. Its causal relation to the disease can be readily proved either by injecting subcutaneously or by feeding small birds with pure cultures. By both methods the disease is reproduced ; by the former, however, owing to the severity of the local lesion, death takes place when the general infection is a t an early stage ; by the latter, there is a complete reproduction of the disease as i t occurs naturally.

( I t must be stated, however, that the above results have only been obtained in the case of small singing-birds. A subcutaneons injection of pure cultures in pigeons causes only a local inflammatory change with superficial necrosis of muscle ; the condition does not spread, nor does it lead to lesions in the internal organs.)

11. EXPERIMENTS ON RODENTS.

Having thus described the relation of this organism to the disease in birds, we have to consider its effect on certain other animals, those which were used for this purpose being rabbits, guinea-pigs, and mice. A special interest attaches to the result of these experiments, owing to the marked difference between the tissue reactions thus produced and those which occur in birds. It may be premised that the animals employed are lower in the scale of susceptibility than small birds, and that we have corresponding differences in the lesions produced. The methods of inoculation were intravenous, intraperitoneal, and subcutaneous injection, except in the case of the experiments on mice, in which the two latter methods only were employed. In all cases the bacilli used for iiijection were taken from a culture on agar of not more than two days' growth. To such a culture about 5 C.C. of sterile normal salt solution was added, and a fine emulsion was made with the growth. From the appearance of the emulsion one can judge so as to have it approximately of the same richness in bacilli


in a l l cases. I n every case the organs were examined microscopically, both by film preparations and by sections ; cultures also were made from them.

1. GuINEA-PIcs-Intraperitoneal injection of fr to 1 ex.-With this dose of the virulent organism a fatal result follows, usually on the fourth to sixth day. The condition found was approximately the same in all cases.

In the peritoneum there is a considerable quantity of serous exudation, which is more or less turbid, and contains siiiall flakes of fibrin here and there. Fibrinous exudation is specially marked in the upper part of the abdomen, over the liver, spleen, stomach, and great omenturn ; the last being drawn up, thickened, and infiltrated. On the peritoneum, at the site of the needle puncture, there is a sniall nodular swelling with caseous centre. On spreading out portions of the mesentery there are often to be seen small whitish scattered points, the largest of those present on the sixth day being about the size of a small pin-head. Such nodules are also present in the omenturn, but are there obscured by the fibrinous exudation. Of the internal organs the spleen shows most change. It is always enlarged, and may measure 1 4 in. in length by 1 in. in breadth. It is of firm, though somewhat friable, consistence, and is studded with numerous small points, which are larger, more distinct, and more yellow in tint the longer the aiiinial has lived. If death has taken place early, e.g. 011 the fourth day, the nodules are very minute, somewhat translucent, and sometimes rather difficult to distinguish with the naked eye. The liver is swollen, congested, and contaiiis a varying number of small nodules scattered in its substance. These are less numerous, less distinctly rounded, and more of a greyish tint than are those in the spleen. They are always most abundant towards the surface. There may be a small quantity of serous fluid in the pleural cavities, and the lungs may show a few hlemorrhages, but I have never found any nodules in them, or in any of the other organs, after iiitraperitoiieal injection. The typical changes which result from intraperitoneal inoculation are accordingly to be found in the peritoneum, spleen, and liver. Cultures made from these parts gave iiunierous colonies of the bacillus in a pure condition. A few colonies are generally to be obtained, also from the blood of the heart, post-mortem, though niicroscopic examination generally fails to detect any bacilli. The peritoneal fluid, when examined microscopically, shows a considerable number of bacilli, partly lying free, partly contained within leucocytes.

Intravenous injection.-The result varies according to the dose. If only one or two drops be injected, the animal may live nine or ten days. If 0.5 C.C. be used, death occurs about the fourth or fifth day. The syniptoms are those of general malaise, and, before death, collapse with marked lowering of the temperature. On post-mortem examination the outstanding changes are to be found in the liver and spleen,

which are studded with a varying number of nodules, these presenting very much the same characters as in the case of intraperitoneal injection, with differences in appearances according to their age. The nodules may be confined t o these organs, though sometimes a few are present in the lungs, where they may be readily felt to be of pretty firm consistence. I have not observed any nodules in the kidneys, or in any of the other organs. If the connective tissue a t the site of inoculation has been contaminated, there occurs a certain amount of inflaniinatory swelling with caseous infiltration. Otherwise there is no change. The bacilli can be obtained in pure culture from the affected organs and also from the blood. Here also, as in the case of intraperitoneal injection, the nodules are restricted in a remarkable way to the spleen and liver.

Sicbcutancous i./z;iection. - If 0.5 C.C. be iiijected subcutaneously, there forms a localised inflammatory swelling a t the site, which gradually increases, and about the tenth day may be fully half an inch in diameter, and of pretty firm consistence. About this time a process of purulent softening, attended by caseation, occurs, and this gradually increases. There is then found to be a collection of soft caseous material surrounded by a fibrous capsule. On the surface ulceration may occur, and the contents become discharged, leaving an irregular ulcer with caseous wall.

The lymphatic glands corresponding to the site of injection also become affected, undergoing a gradual enlargement, with subsequent caseation and softening. Some of the glands may reach the size of a small hazel-nut. These changes are well seen two or three weeks after injection. If the animal be killed a t this time there are often present a few nodules in the liver and spleen, sometimes in the liver only. These vary in size, some reaching nearly an eighth of an inch in diameter, are well circumscribed, and have a sort of caseous centre. I have not found these nodules in any other organs.

Microscopic examination. - Examination of the lesions has been made a t different stages. The following is a summary of the results :- In no case have I found a condition like that in the natural disease in birds, or one corresponding to the conditioii known as tirbcrculose xooglodiqzie. I n all cases the nodules are foci of tissue reaction, followed by degenerative change ; bacilli are usually present in comparatively small numbers, and sometimes it may be impossible to find any.

The earliest nodules, for example those seen in the liver on the third or fourth day after intravenous injection as small greyish-pink points, are found to be small areas in which the hepatic capillaries are blocked with leucocytes, altered red corpuscles, and a few bacilli. The relation of these areas to the distribution of smaller branches of the portal vein can often be well made out. The liver cells in the affected parts become somewhat swollen and homogeneous in appearance, while

PSE UDO-TUBERCULOSIS IN BIRDS. l i 6

their nuclei lose the power of staining, the result being a sort of coagulation necrosis with much hyaline swelling (Plate XVIII. Fig. 4). A day or two later, proliferation of connective tissue and endothelial cells a t the periphery is well seen, this being accompanied by a pro- gressive infiltration of leucocytes. Along with these changes, the necrosed liver-cells, first a t the margin, undergo a process of absorption and disappear. Nodules a t this stage accordingly often show collections of hyaline necrosed liver-cells in the centre, surrounded by a subacute inflammatory zone. Later still, the centre of the nodule becomes a granular amorphous mass, surrounded by a zone in which there are numerous leucocytes, the whole being enclosed by a capsule of young connective tissue. I have not seen any giant-cells, such as are met with in tubercle. Occasionally, when a nodule forms around the portal tract, the cells of the bile ducts enlarge and proliferate, so as to give an appearance somewhat resembling a giant-cell.

I n cases where death has occurred in three or four days the bacilli are found in large numbers, and in their neighbourhood early necrosis is present. They seem t o have a well-marked action on the red corpuscles, which fuse into a homogeneous mass or become fragmented. Remains of red corpuscles are commonly seen in the larger leucocytes. Bacilli also are numerous within leucocytes, many of which can be seen to be degenerated and breaking down. In the later stages-seen, for example, on the ninth or tenth day-the nodules are of larger size and more circumscribed, proliferative change being present a t the margin. Later still the centre becomes entirely necrosed, and a distinct capsule is formed. Changes of the same nature are seen in the nodules in the lungs. The nodules are irregularly rounded areas of consolidation, resulting from catarrhal along with interstitial reaction. I n the centre, the cellular aggregation is very dense, and there also degenerative change is first seen (Plate XIX. Fig. 7). The vessels in relation to a nodule can sometimes be seen to be thrombosed.

The mesentery (from an acute case after intraperitoneal injection) shows numerous small foci of cellular structure, these being present both along the vessels and in the parts betw-een (Plate XIX. Fig. 5). They are composed of aggregations of leucocytes around bacilli, along with desquamsted and proliferated endothelial cells, some of which are degenerated. Similar aggregations may also be seen on the surface of the spleen, etc. I n the peritoneal exudate large numbers of bacilli are found within polynuclear leucocytes. The kidneys may show cloudy swelling, but I have not found any nodules in them.

I may state that the virulence of the organism is well maintained in series of cultures showing a scarcely appreciable diminution after six months. In the case cif one specimen, which had grown in the laboratory for more than a year, I found that it still possessed the same pathogenic action, though in diminished degree. For example, on

In the spleen, corresponding changes are found.

12-JI. . OF PATH.-VOL. V.

176 ROBERT MUIR.

intraperitoneal injection of the dose mentioned, into guinea-pigs, death did not follow till about the tenth day, when there were young fibrous adhesions between the intestines, an extensive caseous infiltration of the omentum, and fewer nodules in the liver and spleen than is the case with recent cultures.

The results are of a corresponding type to those obtained with guinea-pigs, but are of a more chronic nature, the rabbit being distinctly less susceptible than the guinea-pig. On subcutaneous injection, the changes are practically the same as those stated above-a local inflani- matory swelling which caseates and softens, a similar affection of the lymphatic glands, and the presence of one or two caseous nodules in the liver and spleen. The lesions after intraperitoneal injection exemplify the lower susceptibility. About the twelfth day there is little fluid in the peritoneum, and that present is usually free from turbidity. Nodules may be present over the surface of the intestines, especially over the colon, and some of these may reach the size of a very small pea. The mesentery contains similar nodules, usually more numerous, and of smaller size. They are also present in the omenturn. The abdominal lymphatic glands may be swollen, but usually show no distinct nodules or caseation. The spleen is much enlarged, and contains numerous spherical nodules of various sizes, these being pretty firm in consistence. A few nodules may also be found in the liver. No nodules were found in any of the other organs. By intravenow injection a very acute infection may be produced ; for example, if 1 C.C.

of emulsion be injected, death may follow on the fourth day. I n such a case both the liver and spleen contain enormous numbers of nodules, or rather of €oci of necrosis. The spleen may be enormously enlarged, and has a distinctly firm appearance, somewhat like that of a waxy organ. It is filled with small pepper-like pale foci of early necrotic change.

It is unnecessary to describe in detail the microscopic changes, as they closely resemble those in guinea-pigs. Plate XIX. Fig. 8, shows one of the more chronic nodules from the liver of a rabbit. At the margin of the central necrosed area a few giant-cells may be found, but these are of comparatively small size, and are manifestly degenerating. Their presence is probably an indication of the more chronic reaction in these animals.

3. WHITE MrcE.-Intraperitoneal and subcutaneous methods of inoculation were employed. Mruperitoneal injection produces a result closely similar to that in guinea,-pigs, but there is less tendency to the formation of distinct nodules in the serous membrane. In the case of a moderate dose, death usually takes place on about the sixth day. The peritoneal exudate is chiefly clear serum, with slight traces of fibrin over the liver and spleen. Those two organs are extensively involved, the spleen especially undergoing great enlargement, and being studded with minute pinkish-grey points. I n the liver, only

2. RABBITS.-Injections were made by all three methods.

PSE UDO- TUBZR CUL OSIS IN BIRDS. 177

marked congestion may be visible on naked-eye exaniination ; but on microscopic examination numerous minute foci, similar to those described above, may be found. The fluid in the peritoneum and the liver and spleen give, on cultures being made, numerous colonies of the bacilli.

SlcbcutaneoiLs injcctio.n.--l’he results obtained varied considerably, and the changes found in one animal are of special interest, as forming the link between the condition in rodents and that in small birds. Usually there is formed a t the site of inoculation a small nodule, which undergoes caseation and subsequent softening ; and when the animal is killed two or three weeks after the injection, one or two nodules may be present in the spleen or liver. These, however, show little tendency to spread. In one case death followed on the seventeenth day, and both liver and spleen were studded with numerous rounded nodules, about a sixteenth of an inch in diameter, and of pretty firm consistence. One or two nodules of similar character were present in each lung, but the other organs were free. The centre of the nodules showed a small collection of soft, caseous material. We may say that the result appears to depend upon the number of bacilli which may enter the circulation. Another animal died of an acute general infection on the seventh day ; the spleen was enlarged as in the case of intraperitoneal injection, and was speckled with numerous pale grey points of minute size. The liver also showed a corresponding appearance, though the pale points were of smaller size, and less closely packed. The peritoneum was normal, and there were no lesions discoverable in the other organs. On microscopic examination in this case, the changes, as already mentioned, were different from those met with in any of the other rodents. In the liver there were innumerable small masses of bacilli lying in the hepatic capillaries, these masses increasing in size, and displacing the liver cells. Around most of them there was practically no tissue reaction, and the liver cells appeared practically healthy. In some of the larger masses, however, the bacilli were surrounded by a cellular zone -1eucocytic accumulation and young connective tissue proliferation. The central parts of the masses of bacilli were often stained faintly, the organisms there having died out. The condition present is shown in Plate XIX. Fig. 6. The spleen showed numerous masses of bacilli scattered irregularly through its substance, with numerous points of commencing necrotic change, broken-down red blood corpuscles, leucocytes, etc. Though the condition in this case resembled that in birds in the large iiumbers of bacilli present, it differed from it in the fact that here there was more evidence of tissue reaction. It is somewhat difficult to explain the differences in this case from those met with in others, but I think it is possible that in the process of injection a large number of bacilli may have suddenly entered the circulation.

The microscopic appearances in the other cases correspond with

A few can be obtained also from the heart after death.

178 ROBERT MUIR.

those already detailed. In all the larger nodules the central part was a collection of amorphous material, in which no living cells remained, whilst bacilli, usually somewhat scanty, were found at the margin of this material. Around the caseous material there was a dense zone of leucocytes, whilst beyond this again a capsule of young connective tissue was present. All chronic lesions conform more o r less closely to this type.

Feeding experiments.--I have attempted to infect guinea-pigs and white mice by feeding them with cultures, but so far without success. I am making further experiments on this subject.

SUMMARY.

When the bacilli are introduced into the subcuheous tissue of rodents, the lesion may be said, in general terms, to be a subacute inflammatory focus, which undergoes caseation and usually softening. A similar lesion is found in the lymphatic glands related to the part. The rapid necrotic action upon cells, both leucocytes and connective tissue corpuscles, is a well-marked feature. A few nodules may be found in internal organs, but they are usually confined to the liver and spleen. Usiially a fatal result does not follow, though sometimes it may.

Both intravenous and iiitraperitoneal injection give rise to a general affection, the acuteness and extent varying of course with the dose employed. In both modes the outstanding feature is the extensive affection of the liver and spleen with nodules, often to the exclusion of other organs. The nodules are foci of tissue reaction with central necrosis, in which it may be difficult to find bacilli. I n only one instance (in a white mouse) was a condition like that in birds met with. I n the case of intraperitoneal injection, the lesion of the serous membrane is a subacute inflammatory condition, with. the formation of cellular nodules varying in size according to their age.

As compared with tubercle, various differences may be mentioned. In the first place, there is more inflammatory reaction with more extensive leucocytic infiltration. Caseation and softening are also more rapid, the leucocytes also being involved in the process. The result is that the centre of a nodule after a time is rather a focus of caseous pus, and thus differs considerably from the tubercle nodule of corresponding age. The degenerative changes are usually advanced before the proliferative changes are well marked, and the latter are often confined to the periphery of the nodule. Giant-cells are absent, or are imperfectly formed, and a structure like a giant-cell tubercle is not met with. The lesion, in short, resembles rather that in glanders, though in pseudo-tuberculosis caseation is more rapid and more extensive. (This can be well seen on comparing the lesions in the two diseases, after subcutaneous injection of the bacilli in guinea-pigs.) AS the


changes in a nodule progress, the bacilli become fewer in number, and in the chronic lesions it may be impossible to find any. When present, they are to be found at the margin of the caseous centre. I n the early stages of acute cases they can be readily found in the liver and in the spleen, but after a time their discovery may be a matter of considerable difficulty, the difficulty being increased by the readiness with which they become decolorised.

From what has been stated, it will be noticed that the lesions in the two classes of animals-birds and rodents-present considerable differences, which well illustrate the effects of different degrees of susceptibility. In the former, subcutaneous injection produces a rapidly spreading acute inflammation, attended by extensive necrosis of muscle, whereas in the latter the lesion is localised, of subacute or chronic character, and is soon followed by caseation. In the internal organs the differences are even more striking ; for while in birds there occurs an enormous growth of bacilli in masses within the vascular system, attended by comparatively little change in the surrounding tissues, in rodents there occurs a well-marked tissue reaction, accompanied by necrosis of the cells of the organ, whilst the bacilli gradually diminish in number. It is noteworthy, however, that in all animals used, the spleen and liver are the organs in which the lesions specially occur, and very extensive lesions may be exclusively present in them. This fact forms an additional illustration of the susceptibility which diffemit organs possess towards different bacteria.

In the course of my observations on the characters and patho- genetic effects of this organism, I have been struck by the points of resemblance to those of the bacillus of pseudo-tuberculosis of Nocard, Pfeiffer, and others ; and, in view of all the facts, I have come to the conclusion that the two organisms are the same. There can be no doubt that in the case of the great majority of authors quoted above, one and the same organism is described, though there are discrepancies on certain points (vide p. 1 6 1 e t sep.). Preisz, for example, finds that the bacillus is motile, while Pfeiffer and Delbanco describe it as devoid of motility, the organisms in question being undoubtedly identical. Differences in statement are also found regarding the characters of the growth on potato. I may state, however, that regarding these very points, on which the descriptions of various writers disagree, I have either obtained varying results, or had dificulty in forming a definite opinion.

If I am correct in the opinion that the bacillus which causes this disease of birds is the same as the pseudo-tuberculosis bacillus of these writers, the above observations are of some importance, inas- much as they reveal a source of pseudo-tuberculosis hitherto overlooked. Hayem’s observations, though called in question by some, go

180 ROBERT MUIR.

to show that the disease may affect the human subject, but it is quite unknown to what extent this may occur. A knowledge of the lesions produced in animals and of the distribution of the bacilli is, however, an essential in the study of the question.

BIBLIOGRAPHY.

1. DLJ CAZAL AND VAILLARD,

2 LEGRAIN, . . . . . . 3. PREISZ, . . . . . . Centralbl. f. Bakteriol. u. Parasitenk., Jena,

4. KUTSCHER, . . . . . Zt.sc1~r.J Hyg., Leipzig, 1894, Bd. xviii. S. 327. 5. . . . . . . . . Ibid., 1896, Bd. xxi. S. 156. 6. KOTLJAR, . . . . . . Ann. de I‘Inst. Pasteur, Paris, 1894, tome viii.

7. MALASSEZ AND VIGNAL, . Arch. de physiol. norm. et path., Paris, 1883,

Ann. de l’lnst. Pasteur, Paris, 1893, tome vii.

Bull. Aead. de nikd., Paris, 1891, p. 1019. p. 353.

1891, Bd. s. S. 568.

p. 479.

(3) tome ii. p. 369. 8. 3 , 9 , . Ibid., 1884, (3) tome iv. p. 81.

10. ,, . . . . . . Fortschr. d. Met!., Berlin, 1885, Ed. iii. 9. EBERTH, . . . . . . Virrhow’s Archiv, 1885, Ed. c. S. 23.

S. 179 ; and Virc7iow’s Archiv, 1886, .Bd. ciii. S. 488.

Bull. et nibnz. Soc. centr. de nzkl. v&t., Paris, May 1885.

Fortsclzr. d. Med., Berlin, 1886, Bd. iv. S. 713. Ann. de I‘Innst. Pasteur, Paris, 1887, tome xi.

11. NOCARD, . . . . . .

12. MANFREDI, . . . . . 13. CHANTEMESSE, . . . . 14. CHARRIN AND ROGER,. .

15. DOR, . . . . . . . 16. NOCARD AND MASSELIS, . 17. NOCARD, . . . . . . 18. GRANCHER A N D LEDOUX-

LOBARD, . . . . . 19. COURMONT, . . . . . 20. A. PFEIFFER, . . .

2 1. ZAGARI, . . . . .

22. PARIETTI, . . . . . . 33. MBGUIN AND MOSNY, . . 34. HAYEM, . . . . . .

25. PLANCARD, . . . . .

26. PREISZ, . . . . . . 27. DELBANCO, . . . . .

p. 97. Con@. rend. Acad. d. sc., Paris, 1888, tome

cvi. p. 868. Ibid., p. 1027. C07npt. rend. soe. de biol., Paris, 1889, tome

xli. p. 177. Ibid., p. 608. Arch. de nibd. exper. et d’anat. path., Paris,

Conyt. rend. Soc. de biol., Paris, 1889,

‘‘ Ueber die bacillare Pseudotuberkulose der

Ref., Centralbl. f. Bakteriol. u. Parasifenli.,

E d . , S. 577. Ibid., 1891, Ed. x. S. 775. Seniaine nit%., Paris, 1891, tome xi. p. 285,

and Bull. Soc. nzkd. d. hGp. de Paris, 1891, (3) tome viii. p. 378.

“ Des pseudotuberculoses microbiennes,” Montpelier, 1893; ref. Centralbl. f. Bak- teriol. u. Parasitenli., Jena, 1894, Bd. xv. S. 501.

Ann. de l’lnsf. Pasteur, Paris, 1894, tome viii. p. 231.

Beitr. z. path. Anat. u. z. allg. Path., Jena, 1896, Ild. xx.

1889, tome i. p. 203.

tome xli. pp. 215, 513.

Nagethieren,” Leipzig, 1889.

Jena, 1890, Bd. viii. S. 208.

JOURNAL OF PATHOLOGY.--VOL. V. PLATE XVIII.

JOURNAL OF PATHOLOGY.-VOL. V. PLATE S I X .

PSE UDO-TUB3RCULOSIS IN BIRDS. 181

DESCRIPTION OF PLATES XVIII. AXD XIX.

PLATE XVIII.

FIG. l.-Filn~ preparatioii made from the spleen of a bird affected with pseudo-tuberculosis, showing iiumerous bacilli, in some of which polar staining is seen.

Thionin-blue. ( x 1000.)

FIG. 2.-Film prepaiation from young agar culture of the bacillus of pseudo-tnherculosis


PIG. 3.-Section of spleen of a canary affected with the disease, showing the nodules which


FIG. 4. -Liver of guinea-pig, showing an early nodule of pseudo-tuberculosis, fourth day In the centre the necrotic change in

of birds.

are chiefly composed of bacilli, with little change in the surrounding tissue.

after intravenous injection of the bacillus. the liver-cells is seen, leucocyte infiltration especially a t the periphery.

Hzmatoxylin, rubin and orange. ( x 250.)

PLATE XIX.

FIG. 5.-Portion of mesentery of guinea-pig, five days after iritraperitoneal injection of the

( x 20.)

FIG. 6.-Liver of mouse, showing a nodule composed of cellular connective tissue, in which darkly-stained clumps of bacilli are seen ; the more faintly stained centre is a large mass of bacilli in process of degeneration; plugs of bacilli also in the surrounding capillaries. Seven days after subcutaneous injection of the bacillus.

bacillus, showing the small cellular nodules.


FIG. 7.-Section of a nodule in the lung of a guinea-pig, nine days after intravenous injection of the bacillus, showing the necrosed centre and consolidation around.

Hzmatoxylin rubin and orange. ( x 50.)

FIG. S.-Nodnle of pseudo-tuberculosis in liver of rabbit, fifteen days after subcutaneous iujection of the bacillus.

a. Caseous centre. b. Zone of leucocytes, many of which are degenerated, among them some

imperfectly formed giant-cells. c . Zone of young connective tissue. d. Outer zone of leucocytes, chiefly lymphocytes, along with numerous

e. Liver-cells. eosinophiles.

Hzmatoxylin, rubin and orange. ( x 50.)

Documents

On pseudo-tuberculosis, with special reference to pseudo-tuberculosis in birds