ABSTRACTS AND REPORT. 157

that is not demonstrable be held, owing to the difficulty associated with explaining infection on lJhysiological and anatomical grounds, such a blood invasion is of no moment. Such an infection could not be considered as generalisation in the sense of the term as used by Weigert, for by it he indicated the final stage of the disease. The author has found remarkable pronounced lesions in the early stages on a number of occasions. Meat inspection as a practically applied science can only rest upon demonstrable facts.

If, then, the muscle as a rule proves free from organisms while the glands are diseased, and if it can be experimentally shown that infection of the glands is possible without the organism being demonstrable in the blood, these facts must be taken into consideration. (Muller, Zeitscltr. f Fleisch-u.­Milchhy., Vol. XXI!., No .. 4, January 1912, p. 106.)

HEPATIC BLASTOMYCOSIS OF THE GOOSE.

THE term blastomycosis is applied to diseased conditions caused by the multiplication in the solid tissues or in the serous cavities of blasto­mycetes-organisms which are closely related to the yeasts. In the lesions the parasites occur in the form of round or oval cells, which for the most part are discrete. They multiply by a process of budding. Cultivation of qrganisms of this type is easy, and under certain conditions, which are still imlJerfectly known for a number of them, there appear in the cultures certain forms whIch contain a variable number of spores (two to four).

A number of diseased conditions have been met with both in man and in the domesticated animals caused by the blastomycetes. The presence of organisms of this type has been observed in malignant tumours,and some author~ have suggested that they playa part in the causation of cancer.

The authors have discovered blastomycetes on two separate occasions in tatty livers of geese, where they were responsible for somewhat curious lesions.

The livers were obtained from geese that were fat and had been killed at the termination of the period of cramming. Only some fragments of the organ were available for examination, one of which weighed 375 grammes. The lesions were similar in the two cases.

Around the edge of the liver there were fifteen to twenty sacs, varying in size from a small cherry to a nut, and connected with each other by narrow channels. The sacs were sessile, and had large areas of attachment to Glisson's capsule. They did not penetrate into the parenchyma of the liver. The sacs were yellowish-white in colour and fluctuating, and they presented an appearance like a chain of echinococcus cysts along the edge of the liver.

On incision a thick, yellowish-white gelatinous material escaped, and on standing was soon cO:1Verted into a-pus-like liquid. The capsule of the liver was found to be continuous with the walls of the sacs and thickened.

Microscopic examination of the liquid showed that it contained large numbers of small cyst-like structures, which enclosed enormous numbers of the parasites. The organisms were round or oval in shape, discrete or collected into masses embedded in a gelatinous matrix. The parasites were refractile, and provided with a distinct membrane enclosing a nucleated protoplasmic mass. A large number were. observed to be budding, Qne or more little rounded bodies being visible at one or other po'e. The parasites varied in size from 1'5 to 3 microns in diameter.

L

ABSTRACTS AND REPORT.

Histologically the walls of the sacs showed no special features; they were fibrous in structure and fairly thick. The liver tissue was compressed and, showed areas of cirrhosis. No details were obtained by the authors as to whether the geese had shown any symptoms which could be ascribed to the blastomycetes, but it appears to be probable that there were none, since the animals were killed when the process of fattening was complete.

Cultivation .-Glycerinised potato, glycerin agar, and broth were sown with material from the sacs and kept at room temperature (8° to 19°). On the 1>urface of the potato small white colonies made their appearance some days after inoculation. These increased in size and became confluent, forming a creamy white layer over the whole of the potato surface. On microscopic examination this layer was found to be composed of rounded and oval ·elements showing budding, and also rods and short filaments of variable length. The significance of these filaments was not determined. Similar filaments have been described as occurring in cultures of other blastomycetes, but no opinion has been expressed as to their true nature. J'ossibly they represent a mycelium which is unable to develop upon unsuitable media. It is probable that in their saprophytic existence the blastomycetes form a vegetative mycelium and reproductive elements quite different from those found in animal tissues, which have undergone marked simplification owing to their parasitic existence.

Subcultures on glycerinised potato and glycerin-agar were made from the primary culture. The potato was covered with a thick creamy layer in eight days. Growth was slower on the agar, but the characters of the cultures were the same, and microscopic examination revealed the same cellular ·elements and short rods. The authors were able to make out from prepara­tions of this culture that the rods and filaments develop from the rounded elements. A small bud first makes its appearance, which then becomes elongated, but remains attached to the parent cell by a thin filament. This finally breaks through and the rod becomes free.

The organism grows well on other media at laboratory temperature. In glycerin broth white flocculi appear in the upper parts of the liquid on

the second or third day. These gradually enlarge and fall to the bottom of the tube, where they form an abundant deposit. This deposit has the same microscopic characters as the growths on the other media.

On the surface of carrot growth is extremely rapid, and the cultures present the same characters.

Growth is very slow in a I per cent. solution of glucose; the liquid remains limpid and there is no surface film, but a little deposit appears at the bottom ·of the flask.

In broth made from carrot alone, or with the addition of hydrochloric acid (2 cc. of 2 per 1000 HeI in 8 cc. of broth), carbonate of soda (2 per 1000

solution), or tartaric acid solution (2 cc. of a 20 per cent. solution), growth is very rapid. A white film forms on the surface of the liquid on the second or third day, and a thick flocculent layer forms at the bottom of the tube.

Glucose is less favourable for growth than glycerin. The authors were unable to get any of the sporulating forms in their

cultures, nor did they appear in cultures two and a half years old. No growth was obtained on plaster. Inoculations.-Inoculation yielded negative results. Pigeons, guinea-pigs,

and rabbits were inoculated intravenously, intra peritoneally, and intra­muscularly with large doses without result. In two pigeons inoculated intraperitoneally and intramuscularly with a broth culture there were found five months afterwards some small nodules the size of a pin's head in the mesentery and in the muscles. These nodules were yellowish-white in colour,

ABSTRACTS AND REPORT. 159

and contained blastomycetes similar to those in the cultures, but the birds were in perfect health and very fat.

The blastomycetes are saprophytes with a wide distribution in nature. They are found principally on vegetable structures, such as fruit and grains. The two cases described by the authors must have been produced by the introduction of the parasite into the bodies of the geese on the food. There is no evidence to show how the parasites reached the liver or how the lesions were produced.

With regard to the classification of the parasite found, the authors consider that it should be placed in the genus cryptococcus, and suggest the name cryptococcus anseris. (Martin and Daille, Rev. Vet., Vol. XXXVI!., NO.3, 1st March 1912, pp. 129-134.)

DIRECT ARTIFICIAL CULTIVATION OF THE TREPONEMA PALLIDUM.

THE plan adopted by the author for the cultivation of the treponema pallidum from the testicular lesions of rabbits inoculated with syphilis is not applicable to highly contaminated lesions in the human subject. The method referred to was the use of a liquid medium, followed, after adaptation had taken place, by a solid one.

In working out a method of direct cultivation two important points must be taken into consideration, viz.: (1) the nature of the medium, which must be strictly anrerobic, and (2) the power possessed by the treponema of migrating in solid media. The medium successfully employed by the author consisted of 2 parts of 2 per cent. slightly alkaline agar and I part of ascitic fluid or hydrocele fluid to which a piece of sterile animal tissue had been added.

When stab cultures are made in this medium with a liquid containing the treponema and bacteria, the latter remain along the stab, while the former migrate into the substance of the medium and there multiply.

This medium appears to be superior to that devised by Schereschewsky (which contained gelatinised horse serum). because of the liquefaction of the latter by bacteria.

The tubes of medium should be incubated for a few days to determine sterIlity. The author found that the best animal tissue for the purpose was rabbit kidney or testicle, but that other tissues, such as human placenta, sheep's testicle, etc .• could be used.

Small pieces of lesions rich in the parasite, such as chancre. condyloma, or skin papule, should be snipped off. and. after washing in sterile salt solution, should be immersed in sterile salt solution containililg I per cent. sodium citrate. The pieces should then be cut into small fragments and used as follows: One piece is emulsified in a mortar with citrate solution, the emulsion being examined under dark-field illumination to ascertain the richness of the material, and the others are inserted whole into the tubes of culture medium.

The small fragments are forced down to the bottom of the medium by means of a thin glass rod or a platinum loop, and several drops of the emulsion are deposited deeply into the medium. The surface of the medium has about 3 cc. of sterile paraffin oil placed on it to prevent evaporation. Care must be taken to avoid rupturing the medium.

The tubes are incubated for two to three weeks. The tubes usually show a dense bacterial growth along the stab, which

is surrounded by a hazy opalescent zone.