PubMed Health⌕ Search

Biomedical subjects

S Tsubaki

Publications and source records attributed to S Tsubaki.

At least 73 records · Page 4Linked to original sources

Ecology of Rhodococcus (Corynebacterium) equi in soil on a horse-breeding farm.

The ecology of Rhodococcus (Corynebacterium) equi in soil was studied on a horse-breeding farm. R. equi was cultured from soil at a depth of 0, 10, and 20 cm on the six sites of the farm at monthly intervals for 10 months from March to December of 1983. The highest numbers of R. equi were found in the surface soil. The mean number of bacteria in soil samples at every depth increased remarkably from 0 or 10(2) to 10(4) colony-forming units (CFU) g-1 of soil in the middle of April, and later decreased gradually. R. equi inoculated into six soil exudate broths prepared from surface soils at separate sites yielded suspensions with different optical densities, indicating differences in growth. The distribution of serotypes in the soil was similar to that in the horses on the farm. These findings indicated that R. equi could multiply in the soil and flourish in the cycle existing between horses and their soil environment.

Actinomycetales↗

Quantitative aspects of fecal Rhodococcus (Corynebacterium) equi in foals.

Quantitative aspects of fecal Rhodococcus (Corynebacterium) equi in newborn foals for 12 weeks after birth were investigated on two horse breeding farms. R. equi was found in the feces of foals during week 1 of life. The greatest numbers of R. equi were present in the feces of foals during the first 8 weeks of their lives, which coincides with the age when foals are most liable to be exposed to R. equi.

Age Factors↗

Immunoglobulin and specific antibody responses to Rhodococcus (Corynebacterium) equi infection in foals as measured by enzyme-linked immunosorbent assay.

Humoral immune response to intestinal Rhodococcus (Corynebacterium) equi in horses was studied by enzyme-linked immunosorbent assay. Anti-R. equi immunoglobulin M (IgM), IgG, and IgA antibodies were demonstrated in the healthy horse population. Adult horse levels of anti-R. equi IgM and IgG antibodies were reached by 5 to 9 weeks of age in two healthy newborn foals. R. equi was recovered from the foals in the range of 10(3) to 10(4) per g of intestinal contents. A 1-week-old foal was infected with R. equi by mouth daily for 9 weeks. The foal did not show any clinical signs of illness. Anti-R. equi IgM antibody values in the foal increased about 5 to 8 weeks after initial inoculation, similar to the naturally occurring immune response to intestinal R. equi. There were differences among the antibody responses to R. equi in healthy horses, foals with suspected infection, and infected foals. These results suggest that exposure to R. equi is widespread in the horse population and that intestinal R. equi is the most important source of antigenic stimulation for a naturally occurring immune response in horses.

Age Factors↗

Quantitative fecal culture for early diagnosis of Corynebacterium (Rhodococcus) equi enteritis in foals.

Quantitative culture of Corynebacterium (Rhodococcus) equi from feces of 17 foals on a farm (A) with an endemic C. equi infection problem and 26 foals on a farm (B) without the disease in the past decade was done with a selective medium at weekly or monthly intervals from April to August of 1984. Corynebacterium equi was observed in the feces of 16 of 17 foals on farm A, and 19 of 26 foals on farm B. The mean viable count of C. equi in one gram of feces was 4.1 +/- 3.7 (log10) on farm A, and 3.9 +/- 3.4 (log10) on farm B. Corynebacterium equi was recovered from feces of foals as young as two weeks old. Almost all foals at an age between two to four weeks shed the bacteria in the feces. During the observation period two foals showed clinical signs: fever, diarrhea, and cough, at four or five weeks old. At the same time the bacterial count per gram of feces increased from 4 to 7 or 8 (log10). They shed large number of bacteria in the feces and continued to show the clinical signs until death at 10 or 11 weeks old. One of the foals was diagnosed as having had C. equi enteritis and pneumonia by the postmortem recognition of lesions with bacteriological confirmation. The quantitative culture of the feces of foals at weekly intervals after birth on farm A was found to be very useful as an aid in early diagnosis of C. equi enteritis in foals.

Actinomycetales Infections↗

Correlation of in vitro properties of Rhodococcus (Corynebacterium) equi with virulence for mice.

To study the virulence of Rhodococcus (Corynebacterium) equi, seven ATCC strains of different serotypes were tested for their LD50 in mice, clearance of the organism from the lungs and spleen following intravenous or intratracheal inoculation, and in vitro interaction with murine peritoneal macrophages. Strains ATCC 33704 and 33705 were virulent for mice and multiplied in the lungs and spleen, resulting in death of the animal in 5 days. The other five strains were avirulent for mice. The number of bacteria in the lungs and spleen of mice given these five strains decreased immediately. Pulmonary clearance of strains ATCC 33703, 33706, and 33707 was significantly more rapid than that of the virulent strains ATCC 33704 and 33705 12 hr after inoculation. Complete clearance of the avirulent strain ATCC 33707 occurred by day 14, while that of virulent ATCC 33704 and 33705 strains occurred by day 30. The virulent strains ATCC 33704 and 33705 were resistant not only to phagocytosis but also to intracellular killing by macrophages. Strains ATCC 33702 and 33706 were rapidly killed by macrophages although they were rather resistant to phagocytosis. Strain ATCC 33703 was easily phagocytized though resistant to killing by macrophages. The most avirulent strains, ATCC 33707 and 6939, were easily phagocytized and rapidly killed by macrophages. These results indicate that virulence appeared to be related to the ability of the organisms to resist clearance from the lungs and spleen and to resist phagocytosis and intracellular killing by macrophages.

Animals↗

Enzyme-linked immunosorbent assay for diagnosis of Corynebacterium (Rhodococcus) equi infection in foals.

An enzyme-linked immunosorbent assay (ELISA) was used to diagnose Corynebacterium (Rhodococcus) equi infection in foals. In tests done with different antigen-extraction procedures (sodium dodecyl sulfate, sodium deoxycholate, polyoxy-ethylene [9] p-tert-octylphenol, polyoxy-ethylene [9-10] p-tert-octylphenol, sonification, homogenization, and heat treatment at 121 C), Tween 20 was a satisfactory reactive antigen. Using hyperimmune rabbit sera or infected foal sera, we investigated the specificity and the sensitivity of the ELISA with the Tween 20 antigen of the different serotypes or of the isolates. Corynebacterium equi strain ATCC 6939 antigen had the best activity for detecting antibodies to C equi in foals. Sera from 218 healthy horses, 11 healthy foals, 17 healthy newborn foals, a foal with suspected C equi infection, and 5 infected foals were evaluated for antibodies to C equi, using ELISA. The optical density values of 206 healthy horses, 17 healthy newborn foals, and 9 healthy foals were less than 0.1. Infected foal sera, except from foal 3, and serum from a foal with suspected C equi infection had higher optical density values. Using ELISA, specific antibodies against C equi were detected in a naturally infected 6-week-old foal after the foal had a rapid increase in the number of bacteria in the feces and after the initial development of clinical signs of illness at 5 weeks of age. Therefore, ELISA was useful for the early diagnosis of C equi infection in foals.

Animals↗

Use of alpha-naphthyl acetate esterase staining to identify T lymphocytes in cattle.

Peripheral blood lymphocytes (PBL), lymph nodes, and/or spleens from clinically normal cattle were examined for cytochemical staining of alpha-naphthyl acetate esterase (ANAE). Two types of positive-staining patterns in ANAE staining resulted. By a combination of ANAE staining and latex-ingesting test, diffuse ANAE-positive cells were considered as mononuclear phagocytic cells. Using erythrocyte rosettes, erythrocyte antibody complement rosettes, nylon-wool column technique, surface immunoglobulin (SIg) staining, and the ANAE staining technique, granular ANAE-positive lymphocytes were shown to be T lymphocytes. The frequency of T and B lymphocytes in PBL, spleens, and lymph nodes of clinically normal cattle was measured, using ANAE staining and SIg staining. In PBL, 47.7% were ANAE-positive and 26.9% were SIg-positive; in spleens, 22.4% were ANAE-positive and 53.7% were SIg-positive; and in lymph nodes, 38.5% were ANAE-positive and 28.3% were SIg-positive. The frequencies of T and B lymphocytes in PBL, spleens, and/or lymph nodes from cattle with enzootic bovine leukosis (EBL) and cattle with persistent lymphocytosis and in tumor cells from cattle with EBL were measured. When compared with those of clinically normal cattle, PBL, spleens, and lymph nodes of cattle with EBL and the PBL of cattle with persistent lymphocytosis contained numerous SIg-positive cells and few ANAE-positive cells. Tumor cells from cattle with EBL contained 7.3% ANAE-positive and 78.0% SIg-positive cells.

Animals↗