PRELIMINARY INVESTIGATIONS ON COMPLEMENT-FIXING ANTIGENS IN BOVINE TUBERCULOSIS

The principal subject discussed in the present paper is the antigenicities of several kinds of antigens for the C. F. T. in bovine tuberculosis. 1. The antigenicities of next-mentioned human and bovine type of antigens were examined ; Boiling supernatant, bacterial carbohydrate, bacterial protein, ether-extracted, ether-alcohol-extracted lipoids which were prepared from the dried bacilli preliminarily inactivated by temperature of 70℃ and 100℃, and tuberculin. 2. The boiling supernatant antigens from the human type bacilli by low temperature inactivation and from the bovine type bacilli inactivated by both low and high temperature, equally indicated very good antigenicity and specificity. 3. Bacterial proteins possess considerably good antigenicity, however, their reaction ranges was not so wide but that they easily lost their antigenicity by comparative low dilutions. 4. Tuberculin also indicated excellent antigenicity but the nonspecific reactions were sometimes recognized. 5. Ether-extracted lipoid antigens are not recommendable ones on account of their weak antigenicity, especially in the one from bovine type, and on account of non-specific reactions. 6. Bacterial carbohydrate antigens do not have any antigenicity at all

STUDIES ON TOXOPLASMOSIS I : AN OUTBREAK OF TOXOPLASMOSIS AMONG HARES (LEPUS TIMIDUS AINU) IN SAPPORO

A severe outbreak of toxoplasmosis was observed in a hare ranch (Lepus timidus ainu) in Sapporo. The data obtained are summarized as follows : 1. Outbreak occurred in rather warm season during the period of August 15〜September 11,1957. 2. Out of 13 hares including 8 adult and 5 about 1〜2 months old, total 8 had succumbed one after another within a short period. 3. Infection rate was higher in adult (7 out of 8-87.5%) than in young (1 out of 5-20%). No subclinical or latent infections were recognized clinically. 4. Disease runs a very acute course without representing any nervous symptoms. 5. Autopsy findings exhibited the necrotic changes completely in coincidence with those already reported. 6. Parasites were demonstrated heavily in the liver, spleen and mesenteric lymph node by the GIHMSA stained smear. The isolation of the parasites was successful in 1 case after 4 successive inoculations to mice. 7. The present observation in hare was the first reported in Japan

BACTERIOLOGICAL STUDIES ON STREPTOCOCCI FROM BOVINE UDDER I. : SEROLOGICAL AND BIOCHEMICAL OBSERVATIONS ON GROUP-B STREPTOCOCCI AND A GENERAL DESCRIPTION OF STREPTOCOCCI FROM BOVINE MILK IN HOKKAIDO

Bacteriological studies on streptococci, especially on Str. agalactiae isolated from bovine milk in Hokkaido during 1955 and 1956 were described. The data obtained are summarized as follows : 1. A total 3508 quarter milk samples derived from 886 cows were examined from the view point of a study of streptococci. Incidence of streptococci in bovine milk in Hokkaido was 27.0% in cows (239 out of 886), and 11.9% in quarters (418 quarters out of 3508). 2. These streptococci were demonstrated from average 69.2% of mastitis quarters and 51.4% of quarters which secreted the abnormal milk, though it was demonstrated from normal milk in 18.8%. These percentages are widely diverse by farms. 3. Isolated strains of streptococci to the number of 319 were divided into Str. agalactiae ; 126 (39.5%), Str. uberis ; 80 (25.1%), Str. bovis ; 34 (10.7%), Str. faecalis ; 17 (5.3%), Str. lactis ; 12 (3.8%), Str. dysgalactiae ; 8 (2.5%), Str. acidominimus ; 5 (1.6%), Str. pyogenes ; 4 (1.3%) and others. 4. Eighty percent of Str. agalactiae and Str. dysgalactiae strain, 70% of Str. uberis strain and about 90% of Str. lactis strain were derived from mastitis or abnormal milk. Contrary to this, more than 50% of Str. bovis or Str. faecalis were isolated from normal milk. 5. One hundred forty-five strains which reacted with group-B serum were serologically typed into type II ; 114 (78.6%) and type III ; 1 (0.7%) by precipitation reaction following LANCEFIELD. The author could not demonstrate any type specific carbohydrate antigens in the remaining 30 strains (20.7%) examined. 6. Some experiments were performed by agar gel diffusion method on the strains lacking type specific carbohydrate. Discussion was offered. 7. Survey on the distributions of these sero-types among cows in 4 farms showed interesting results characteristic to each farm. 8. Biochemical properties, especially sugar fermentation reaction were examined. According to this there seem to be 3 types ; the salicin-negative- and trehalose-positive type, the salicin-positive and trehalose-negative type and the both-positive type. One biochemical type of Str. agalactiae tends to be detected in 1 farm. 9. Str. agalactiae produce acid from galactose, mannose, levulose, sucrose but not from xylose, rhamnose amygdalin, raffinose, inulin, sorbitol and mannitol without exception. With the characters of hydrolysis of sodium hippurate, non-splitting of esculin and production of ammonia from arginine, these fermentation reactions seem to be invariable by strain. 10. The strains which manifested the above mentioned biochemical characters all reacted promptly strong positive with group-B sera by precipitation

STUDIES ON TOXOPLASMOSIS V : COMPLEMENTAL OBSERVATIONS ON THE TISSUE CULTURE METHOD, ESPECIALLY ON THE EFFECT OF THE NUTRIENT FLUID UPON THE INVASION AND MULTIPLICATION OF THE ORGANISMS

Following the previous report, complemental observations were carried out to ascertain the suitable cultural conditions including the effects of the nutrient fluid upon the invasion and multiplication, the ultimate aim being to promote the utility of tissue culture in isolation of the organisms. Throughout the experiments HeLa cells and T. gondii strain RH were used. Data obtained are summarized as follows : 1. Good invasion occured very frequently when Hanks' balanced salt solution and solutions devoid of divalent ions such as Mg, Ca, were employed. However, there seems to be no significant differences in the ratio of invasion due to the culture fluids tested-H (Hanks), HL (Hanks plus lactalbumin hydrolysate), HLS (HL plus serum), YLE (yeast extract, lactalbumin hydrolysate and Earle's balanced salt solution) and YLES (YLE plus serum), and organism numbers more than 90% may invade the host cell 24 hours after inoculation (fig. 1). 2. The organisms which were suspended in the culture fluid devoid of protein substances tend to die in rather short period and could not multiply well in cells when cultured by these proteinless nutrients (Fig. 3 A, Table 1). Accordingly, the nutrients must be replaced by ones containing protein after 5〜7 hours when the media such as Hanks were employed in inoculation (Fig. 3B). 3. Addition of bovine sera to the concentration of 0.5〜1.0% seems to be quite enough to cause heavy growth of T. gondii if the medium HL is used for the culture nutrient. 4. PH values ranging from 6.5 to 8.0 do seem not to show any effects on the invasion ratio of the organisms. However, in the more acid or alkaline sides (6.0,8.0) poor multiplication was always observed and especially in acid side, elution of the soluble complement-fixing antigenic substances was also scarce (Fig. 2). 5. The volume of the suspension of the inocula seemed to have no significant relations with the yield of the organisms when the inoculated bottles had been left overnight for the invasion (Fig. 7). 6. To cultivate a small number of the organisms in material, frequent changes of the nutrient fluid every day or every other day during the first week are very necessary (Fig. 6). 7. Clear proofs of the multiplication of the organism numbers of 2.5～5.0×10^3 in the materials were observed by counting method when the tests were performed with due consideration of the above described points (Fig. 8)

STUDIES ON TOXOPLASMOSIS II. : SOME OBSERVATIONS ON STRAIN "HT" WHICH WAS ISOLATED FROM A HARE (LEPUS TIMIDUS AINU) IN SAPPORO

The hare strain HT of Toxoplasma gondii which was reported in the previous paper was tested immunologically and also pathogenicity for the laboratory animals and was examined in comparison with known toxoplasma strain RH. The data obtained are summarized as follows. 1. The strain HT indicated high virulence for mice and death occurred generally in 5∿6 days after intraperitoneal inoculation, almost the same as the RH; however the number of parasites in mouse peritoneal fluid was very small in comparison with the strain RH. 2. For rabbit and guinea pig, the strain showed very low virulence. In rabbit, even resulting from intracerebral inoculation, only fever reaction occurred and the general conditions ran always normal afterward. 3. Immunologically, by the use of cross complement-fixation test, this strain was identified as Toxoplasma gondii. 4. At first, the isolated strain HT had revealed no reactivity in vitro in case of dye test with the rabbit sera infected with the strains HT and RH, however the isolated strain had showed the ability to produce cytoplasm-modifying antibody reacting with the antigen RH. After about more than 50 mouse passages, the strain revealed reactivity in vitro the same as the strain RH. The above facts must be remembered for serological identification of the newly isolated strains by the dye test

SALMONELLA TYPES IN ANIMALS IN SAPPORO II

A survey on Salmonella distribution was conducted in apparently healthy animals in and around Sapporo. The data are summarized as follows : 1. Salmonella was detected in 11 cases out of 355 (3.1%) dogs in Sapporo. Specimens were serological1y typed into 4 S. enteritidis, 6 S. typhi-murium and 1 S. thompson. S. thompson from dog is the first finding in Hokkaido. 2. From foxes, total 25 cultures of Salmonella were recovered in 19 cases out of 119 (16.0%). Sixteen of them were S. bareilly. Furthermore this is the first isolation from fox in Japan. The others are 6 S. typhi-murium and 3 S. enteritidis. Some cases showed incidence of 2 types. 3. No Salmonella was detected from 44 minks. 4. These Salmonella from dog and fox were detected from the mesenteric lymph node (70%), liver (30%), caecum contents (30%), spleen (20%) and kidney (3.3%). It is noticeable that no Salmonella was detected from the bile samples

SALMONELLA TYPES IN ANIMALS IN SAPPORO

Some investigations on Salmonella distribution in several animal species in and around Sapporo have been performed. Data are summarized as follows : 1. Eleven Strains of Salmonella organism in total were isolated from 1,233 cases of animals mainly including cattle, hog, dog, fox etc. 2. Salmonella types found were S. enteritidis (5) from cattle, dogs, fox and cat, S. typhimurium (3) from dog, fox and rat, one each S. thompson from hog, S. cerro from fox and S. amager from mink. 3. The isoiation of Salmonella cerro and Salmonella amager were the first to be made in Japan. 4. Besides Salmonella, 4 Bethesda-paracolon group organisms were also isolated from cow, hogs and fox