ABSTRACTS. 12 7

the filtrate is quite germ-free. The filtrate can be preserved for several months by the addition of toluol. (Naoshi Nitta. Bulletz'n oj the Central Veten'nalY Medical Association of Japan, No. I, April 1918.)

ULCERATIVE LYMPHANGITIS.

AT the request of the Consultative Veterinary Committee, the Military Research Centre, instituted at the Alfort Veterinary School towards the end of 1917, has carried out under the successive direction of MM. Panisset and Send rail a series of experiments in connection with the different forms of lymphangitis occurring in the horse. In addition to very valuable information regarding epizootic lymphangitis, which was mainly the result of investigations carried out by M. Boquet, and most {)f which has already been published, the Centre has furnished a series of reports which are summarised in the present article.

Nature of the Disease. - Before undertaking the therapeutic study of ulcerative lymphangitis it was considered necessary to determine its nature with certainty, since in the course of the war doubts were expressed as to the role of the Preisz-N ocard bacillus. As a matter of fact, the microscopic search for the bacillus often has a negative result. Sometimes the bacilli are very scanty and seem to have unusual characters. One may examine several preparations from a nodule without discovering a single bacillus. When the material for examination is taken from an open lesion the presence of secondary invaders puts difficulties in the way of diagnosis.

In order to determine the actual cause of the dis~ase, attempts were made to obtain cultures from materials taken from recent unopened lesions.

Even in cases where microscopic examination fails, the Preisz-Nocard bacillus can always be cultivated in Martin's broth from the lesions. The organism is so virulent that a very small number of bacilli may set up extremely acute lesions; and, since the Preisz - Nocard bacillus is always found in a state of purity when the material is taken from recent unopened lesions, it is justifiable to conclude that it is the actual and only cause of ulcerative lymphangitis.

The bacilli possess the troublesome property of being able to live in a latent condition in the body. They may be found locally in certain lesions which have apparently been cured for a long time, and it is this fact that explains the relapses which often occur at long intervals.

On the other hand, the bacilli, at least in ordinary conditions, have very little power of spreading in the tissues. The lymphatic glands which receive lymph from a diseased limb generally escape attack, and in any case do not contain the bacilli. This absence of adenitis is a point of value for dis­tinguishing between ulcerative lymphangitis and epizootic lymphangitis.

Experimental Transmission. - The constant and exclusive presence of N ocard's bacillus in recent lesions having been firmly established, it remained to reproduce the disease by inoculation with that organism.

At the outset it was thought that it would be necessary to use appreci­able quantities of culture in order immediately to overcome the resistance of the animal. Three horses each received under the skin I cc. of a forty­-eight hours' culture in ordinary broth. One of the animals was inoculated in the neck and the other in the hollow of the pastern. The result was the formation of large abscesses. which burst but were not accompanied by persisting lymphangiti~. In one of the horses, however, thirty - two days after apparent healing of the lesion, three ulcers were found to have formed in the hollow of the pastern, near the point of inoculation. A

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fourth also appeared on the fortieth day, and showed no tendency to cicatrisation. The first attempt to transmit the disease was thus successful.

In order to make the experiment approach more closely to the condi­tions of natural infection, it appeared logical to try the effect of very small doses. On the other hand, having regard to what is known in many other diseases, it appeared possible that powders, or substance capable of exert­ing a negative chemiotaxis, or even accessory bacteria, might favour the development of the disease.

Two subjects were therefore injected in the hollow of the pastern, one with -~ cc., and the other with I cc. of a young culture in broth diluted to one in ten, and mixed with some powdered charcoal. A third received ~ cc. of culture, diluted with ten volumes of a I per cent. solution of lactic acid, and in a fourth subject oil of vaseline was used as a favouring agent. In the case of a fifth subject, a loopful of culture was simply diluted in 10 cc. of normal saline, and I cc. of this mixture was used for inoculation.

The first two horses developed a large swelling, which burst after four days, leaving an ulcerating wound about 3 cm. in diameter. In the second horse, which received a small dose, the lymphangitis appeared to evolve in the usual form. In the horse which had received the oil a slough separated. leaving a large ulcer. The lactic acid appeared to have to a large extent diminished the virulence of the bacilli, as in that case nothing followed except diffuse swelling.

The most demonstrative results were obtained with Horse NO.5, which had been given only a very small dose of culture. On the thirty-second day two small abscesses burst on the inner surface of the metacarpus; on the thirty-seventh day two others on the inner surface of the fetlock; and on the forty-third day, a fifth at the lower part of the hock. This experi­ment was, therefore, very conclusive, inasmuch as the inoculations all caused the formation of lesions resembling those of the natural disease, including local engorgement following the course of the lymphatic vessels. It was therefore concluded as a result of this experiment:-

(1) That the Preisz-Nocard bacillus is the actual specific cause of ulcerative lymphangitis of the horse; and

(2) That it is possible to reproduce the disease experimentally hy locar inoculation with very small doses of young culture, associated preferably with some inert body su~h as charcoal;

(3) That the true symptoms of lymphangitis usually do not appear sooner than a month after the virulent injection.

Contagz'osz'ty.-lt was thought of interest to determine to what extent ulcerative lymphangitis is contagious under ordinary conditions. With that object a number of healthy animals and of animals suffering from epizootic lymphangitis were kept for a very long time in stalls previously occupied by the subjects of ulcerative lymphangitis, and even on the litter which had been soiled by the latter.

It was not found possible to transmit the disease in this way in any case, and it therefore appears justifiable to conclude that there is little danger of direct contagion under ordinary conditions. No doubt maceration of the legs in mud plays a predisposing role by favouring the penetration of the bacilli. This notion has some importance, because it demonstrates that there is no danger of the disease heing spread by the return of a horse suffering from ulcerative lymphangitis to his own stable and ordinary work. The same cannot be said of epizootic lymphangitis, which exhibits the power to spread by contagion much more clearly.

TherapeuHcs.-While the Committee were carrying out these researches on the nature and mode of transmission of ulcerous lymphangitis it undertook a

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series of experiments in order to verify the curative value of some methods already recommended, and in case these failed to search for some truly efficacious method of treatment.

The therapeutic agents which had been recommended included, (I) mallein, tuberculin, rivoltin, etc.; (2) specific vaccine; (3) chemical sub­stances; (4) sera; (5) surgical methods.

Mallein.-Several army veterinary surgeons, having nothing else at hand, had the idea to use mallein in order to excite phagocytory action in the lesions, and they sometimes obtained interesting results. The doses em­ployed varied from 5 to 10 cc. of dilute mallein, a quantity equivalent to icc. of strong mallein. The patients were given from four to six sub­cutaneous injections with intervals of two days. The wounds were simply cleansed by the methods commonly employed.

This method of treatment was tested according to the precise conditions indicated by those who first employed it, and from these tests it appeared to be permissible to draw the following conclusions: Injections of mallein have an undoubted effect on the progress of the disease. This action is manifested first by the production of a very distinct phase of diminished resistance, with slight increase of temperature, a re-awakening of suppuration, and the development of new "buds," sometimes even in old cicatrices. This phase of lowered resistance is not reproduced by successive injections; generally it lasts up to the fourth day of treatment, and then is succeeded by a phase of improvement. Unfortunately, this improvement is not very marked; some of the lesions cicatrise, but relapses nearly always follow.

Larger doses injected intravenously have no better effect. The phase of diminished resistance is shorter and sometimes more decided, and it is reproduced by successive injections; but any therapeutic effect remains doubtful, and, as a rule, new buds make their appearance after an apparent improvement.

Tuberculz'n.-Tuherculin injected in the same conditions as mallein gave exactly parallel results. In both cases the effect is merely a transient stimulus to the lesions, followed by a phase of marked but transient leuco­cytosis, without any retardation of the development of the disease.

Rivoltin.-The autolysate of the yeasts of beer, which has been recom­mended for the treatment of epizootic lymphangitis, had no effect when tried in cases of ulcerative lymphangitis.

Specific Toxin.-The Preisz-N ocard bacillus, which presents many points of analogy with the diphtheria bacillus, like the latter, forms a very active toxin. It was therefore indicated to try the effect of this toxin, either by employing the products of external secretion-that is to say, the soluble poisons which are found in a broth culture (exotoxin), or by using killed bacilli themselves (endotoxin). '

At first treatment with the soluble toxin appeared to be of some benefit. The negative phase was no greater than that caused by mallein and tuberculin. When 20 cc. of the toxin was injected subcutaneously every three or four days pretty rapid cicatrisation of the suppurating lesions took place, but relapses occurred.

Vaccines.-Different kinds of vaccine have been prepared from cultures of the Preisz-Nocard bacillus, killed either by ether or by heat.

Cultures killed by ether, like the toxins, had beneficial but transient results. When injected under the skin they have the inconvenience of provoking the formation of small necrotic ulcers. In order to avoid this result the injections were made into the muscular masses of the croup; and, although no ulceration followed, the injection produced a pronounced inflam­matory swelling in the region, which became hot and painful and caused intense lameness.

Here again the injection was followed by a phase of diminished resistance, H2

130 ABSTRACTS.

manifested at first by an increase of temperature, and later by the develop­ment of fresh lesions, which afterwards tended to disappear gradually, but relapses soon occurred. The vaccine prepared with ether was found to have no real specific action, but simply a stimulating general effect like the other antigens previously mentioned.

A last trial of vaccine practised according to a new technique, which con­sisted in inoculating increasing doses, viz., I cc. the first week, 2 cc. the second week, and 5 cc. the third week, had no better results. Each injection was followed by a phase of slightly diminished resistance, succeeded by some improvement in the lesions; but control subjects which received simple hygienic care were cured in the same proportion as the horses that were treated with vaccine. Moreover, some animals which were treated in a veterinary hospital by a similar method, and afterwards sent as cured to the Research Centre, showed very decided relapses, proving the very slight value of such treatment.

Cultures killed by heat and live cultures were not found to be more efficacious.

Chmzica! Substances.-It was thought of interest to verify the value of various chemical substances which had been previously recommended, or which seemed to conform to certain theoretical indications .

. An English author has recOlpmended as very efficacious the following mIxture :-

Sulphate of copper 30 grammes Carbonate of ammonia 30 II

Water 560 II

Ammonia 4 " This caused some local improvement, but not a definite cure. A I in 200 solution of sulphate of copper injected into the veins in pro­

gressively increasing doses of from 30-60 centigrammes of the sulphate had no decided result. The treatment did not cause any general reaction or coagulation of blood, but the lesions in the horse thus treated showed no difference as compared with those in control subjects.

Arsenobenzene of copper when tested in the same conditions was found to be without any beneficial effect on the disease.

Iodised oil in the strength of 40 per cent. can be injected with impunity into the veins in doses of 5 grammes. Such injections are fairly well tolerated, provided one takes the precaution to strengthen the heart by a simultaneous injection of 10 cc. of camphorated ether under the skin. The only effect produced is some rise of temperate and a little swelling at the point of injection. In spite, however, of an apparent improvement, this treatment does not kill the bacilli throughout the body, for, in four out of six subjects that appeared to be cured, relapses occurred.

Doses of I C;C. of a 3 per cent. Lugol's solution of iodine, introduced by puncture on the surfaces of the diseased limb, proved of no benefit. On each occasion the treatment was followed by a pronounced swelling of the leg, which disappeared after two or three days. Some improvement in the lesions occurred, but they were not curec;l.

The injection into the veins every two days of I gramme of tartar emetic in 50 cc. of normal saline solution was also without beneficial result.

On account of the sensitiveness of the Preisz-Nocard bacillus to acids, it was thought well to try a local treatment based on the employment of these substances. An ointment composed of I of salicylic or lactic acid in 9 of lanoline and vaseline in equal parts was applied to the ulcerating lesions. The ointment had no caustic effect and did not harden the tissues, but the therapeutic results were mediocre. The shallower ulcers cicatrised pretty quickly, but the others improved only slowly. The treatment had the disadvantage of increasing the suppuration during the hot season.

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Upon the whole nothing beyond very partial results were obtained by treatment with chemical substances, and the results did not justify the recommendation of anv of them.

Serum.-The specific character of ulcerative lymphangitis being clearly established, it was natural to try the effect of serum treatment.

Two horses that appeared to have recovered were given at the outset injections of the toxin of the bacillus (filtered cultures in Martin broth) in increasing doses, and afterwards living cultures. One of the two died from renal lesions. The other, after five months of immunisation, had a recrud­escence of the disease in some of the old lesions, and it finally died from the disease in a generalised form.

The attempt to obtain a serum thus failed completely, and it showed that it is not possible to vaccinate against the Preisz-Nocard bacillus, since animals that have received large and progressively increasing doses of toxin and of virus are not protected against the development of specific lesions. In reality, this result might have been foreseen, for up to the present time one does not know any efficacious vaccine against a chronic disease, and from every point of view ulcerative lymphangitis must be regarded as a slowly evolving affection.

In view of this impossibility of protecting the serum-producers themselves against the disease, the attempts at serotherapy were abandoned. Moreover, it was found that the serum of the subjects considered to be hyper-vaccinated had no effect on the bacillus, either locally or at a distance_

The polyvalent serum of Leclainche and Vallee, used for the local treat­ment of ulcers, on many occasions gave interesting results. It facilitates cicatrisation, either by creating a favourable medium or by its action on secondary agents. It cannot, however, be said to be an infallible method of treatment, though it is a valuable adjuvant, especially in those cases in which streptococci are also present, as is the rule in open lesions.

Htemotherapy.-Treatment by the injection of blood from subjects believed to be cured did not give the positive results which some authors have claimed for it. By this method, as by many others, improvement may be obtained, and some apparent cures; but, if one takes care to use animals presenting the disease at the same stage as controls, it is not difficult to perceive that the number of recoveries is as great among the latter as among the animals treated.

Surgical.-Treatment by Bier's method, using passive congestion pro­duced by means of a rubber band, was tried in a number of cases, either by itself or in combination with interstitial iodine injections, but without any apparent benefit.

Cauterisation of the lesions, which is of great value in epizootic lymphan­gitis, especially in the case of lesions situated on the trunk, was found to be of little use in ulcerative lymphangitis, in which the lower parts of the legs are most commonly attacked.

To sum up, it may be said that the different methods of treatment tried were found to have only a palliative effect. They aid recovery in the slighter forms of the disease, but are almost without effect when the lesions are extensive.

Dzagnosis by Specific Reactions.-It was of interest to determine whether ulcerative lymphangitis is a single pathological entity, and, in the contrary case, if it was possible to distinguish the different types of Preisz-Nocard bacillus by their biological reactions.

By injecting into the subpalpebral connective tissue I cc. of a glycerine extract of a pure culture of the bacillus there was produced after twenty-four hours a diffuse swelling of the region, firm to the touch, often very extensive, and reaching as far as the malar crest. The formation of this sensitive swelling is very characteristic. Along with it there is frequently observed

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conjunctivitis, lacrymation, and a more or less abundant formation of muco­pus at the inner canthus. This reaction, however, cannot be considered absolutely specific, for even in healthy subjects the injection produces an <:edema, which, although it is often painless and rapidly disappears, is never­theless capable of being confounded with the genuine reaction. The injection does not cause any disturbance of temperature either in diseased or healthy subjects. At present it would therefore be premature to recom­mend this method of diagnosis, which until it has been further improved must remain a laboratory test.

In reality, however, the diagnosis of ulcerative lymphangitis rarely presents any difficulty. Noone who has seen a case of this intractable disease can afterwards have much difficulty in recognising it. (XXX. Revue Generale de MM. Vet., Tome XXVII!., 1919, p. 233.)

THE BIOLOGY OF PSEUDO-ANTHRAX BACILLI AND THEIR DIFFERENTIAL DIAGNOSIS.

THE so - called pseudo - anthrax bacilli have recently acquired a great importance owing to the fact that anthrax-like diseases in the human subject have been ascribed to organisms of that class. Since pseudo-anthrax bacilli are widely distributed and may in certain circumstances be con­founded with true anthrax bacilli, the differential diagnosis of these organisms becomes important. The author in his researches endeavoured to determine whether clear biological distinctions could be relied upon for diagnosis, whereas in previous investigations reliance was mainly placed on morpho­logical characters. The immediate impulse to the investigation was given by the discovery of pseudo-anthrax bacilli in a pig. The an thor's conclusions are summed up as follows :-

So-called pseudo-anthrax bacilli were cultivated from the muscular tissue of a pig which had been slaughtered while suffering from a disease with anthrax-like symptoms. The case acquired special importance owing to the fact that the slaughterman by whom the pig was killed became ill and died after exhibiting the clinical symptoms of anthrax. Owing to the fact that pseudo-anthrax bacilli are widely distributed in nature and especially in animal products (flesh meal, bone meal, wool, etc.), and frequently present an extraordinarily close morphological resemblance to anthrax bacilli, it is possible to confound them with the latter. This morphological resemblance is sometimes so great that it becomes absolutely impossible to distinguish by microscopic examination between the one and the other, or reliance has to be placed on quite trivial differences.

In young cultures pseudo-anthrax bacilli are generally motile, but they usually become motionless when spore formation begins. It is especially the single free bacilli and the short jointed filaments that exhibit motility, whereas the longer threads are generally motionless. Strains may, however, be met with that are more or less motile. Contrary to what is the case in the Bacillus anthracis, no stainable capsule is demonstrable in pseudo-anthrax bacilli. .

In pseudo-anthrax bacilli spore formation is much more abundant and energetic than in anthrax bacilli, and in agar cultures it is generally complete within twenty-four hours. The spores are larger and more rounded than anthrax spores.

On agar plates the surface colonies of the pseudo-bacilli showed the medusa arrangements less distinctly than anthrax colonies, and at the edge the outrunners are less numerous and less regular in shape. In corre-