SummaryThree cytopathic rotavirus strains were isolated in MA104 cells from faecal specimens of pediatric patients with acute gastroenteritis. Pre-treatment of virus with trypsin and incorporation of a small amount of trypsin in maintenance medium were important for establishment of the strains in these cells. The isolates were antigenically closely related with strain Wa of human rotavirus and had some antigenic relationship with strain Lincoln of bovine rotavirus. , Numerous attempts to propagate human rotaviruses to high titer in cultured cells have failed (1--3, 7, 8, 1t). Recently, WYATT et al. (12) adapted type 2 human rotavirus (strain Wa) to grow to relatively high titer in primary cultures of African green monkey kidney cells after 11 passages in newborn gnotobiotic piglets. In the present study we isolated 3 strains of human rotavirus in cultures of MA 104 cells, a stable cell line derived from embryonic rhesus monkey kidney.Faecal specimens were obtained from pediatric patients who had whitish, watery diarrhea, vomiting and slight fever and recovered in 3 to 5 days. The faecal specimens were collected the day after the onset of the disease and stored at --80°C until use. All the specimens were shown by electron microscopy to contain rotavirus particles when examined by the method described previously (9).Cultures of MA104 cells were prepared in 110 × 10 m m tubes. The growth medium used was Eagle's minimum essential medium (MEM) containing 10 per cent tryptose phosphate broth (TPB) (Difco), 10 per cent calf serum, 100 units/ml penicillin, 100 ~g/ml streptomycin and t ~g/mI fungizone, and the maintenance medium (MM) was MEM containing 10 per cent TPB, 0.5 per cent sodium glut,-0304-8608/81/0069/0155/$ 01.20

A sudden outbreak of epidemic diarrhoea of piglets occurred in Japan, the principal features being watery diarrhoea, dehydration and high mortality in newborn animals. The microscopical lesions were villous atrophy in the small intestine, the villous enterocytes being vacuolated and cuboidal in shape. The villus-crypt ratio was severely reduced, varying from 1:1 to 3:1. Transmission electron microscopy showed numerous coronaviruses within the cytoplasm of enterocytes and among microvilli. Specific antigens of porcine epidemic diarrhoea (PED) virus were detected in the cytoplasm of enterocytes by the streptavidin-biotin (SAB) technique. Infected cells, which were most abundant in the villous epithelia of the jejunum and ileum, were present in small numbers in the large intestine, the crypt epithelia, the lamina propria and Peyer's patches. The study suggests that the SAB technique is useful for the diagnosis of PED.

Akashi, H., Inaba, Y., Miura, Y., Tokuhisa, S., Sato, K. and Satoda, K., 1980. Properties of a coronavirus isolated from a cow with epizootic diarrhea. Vet. Microbiol., m,A coronavirus (Kakegawa isolate) isolated from a cow with epizootic diarrhea was grown in BEK-1 cells and examined for biophysical and biochemical properties. The Kakegawa isolate was able to replicate in the presence or absence of 5-iodo-2'-deoxyuridine, indicating that its viral nucleic acid was RNA. It was highly sensitive to ether and chloroform, and moderately sensitive to trypsin and heat. It was, however, readily stabilized by treatment with cation at 50°C for 1 h. Its infectivity was slightly reduced at pH 3.0. The virus passed through a membrane filter of 200 nm pore size, but not through one of 100 nm pore size. The buoyant density of the virus was determined in a sucrose density gradient. The peak of infectivity and hemagglutinin activity was found at a density of 1.182. Neutralization and hemagglutination inhibition tests showed a close serological relationship between the Kakegawa isolate and the American strain of calf diarrhea coronavirus.0378--1135/80/0000-~000/$02.50

Antigenic relationships between the Wa, Hochi, Ito, Hosokawa and Nemoto strains of human rotavirus, the SA-11 strain of simian rotavirus, the Lincoln strain of bovine rotavirus, the OSU strain of porcine rotavirus and the R-2 strain of lapine rotavirus, which were all established in cell culture, were studied by neutralization (NT) and immunofluorescence (IF) using guinea pig antisera against these strains. The strains could not be distinguished by indirect IF staining of infected cells because of marked cross reactions. On the other hand, the NT test clearly distinguished these strains, although there was a one-way antigenic relationship between some of them. The five strains of human rotavirus were clearly distinguished from each other by NT test. These human strains exhibited much higher homologous titers than heterologous titers, the former being at least eight-fold higher than the latter. The presence of at least five serotypes of human rotavirus was indicated.

Electronic records from the California Animal Health and Food Safety Laboratory System between January 1, 1991, and June 30, 1998, were reviewed, and data on 593 Salmonella serotypes isolated from feces or intestinal contents of adult dairy cattle with diarrhea were analyzed to determine spatial and temporal clustering. A statistically significant clustering in space was detected with 10 (Typhimurium, Montevideo, Muenster, Anatum, Give, Menhaden, Kentucky, Agona, Derby, and Newport) of the 13 serotypes examined. Significant temporal clustering was also detected with 10 serotypes (Typhimurium, Montevideo, Muenster, Anatum, Give, Newbrunswick, Menhaden, Kentucky, Derby, and Newport). Six serotypes (Anatum, Menhaden, Montevideo, Muenster, Newbrunswick, and Newport) were significantly spatially and temporally clustered. A difference in temporal and spatial distribution patterns of some serotypes associated with diarrhea in adult dairy in California was found. Knowledge of the specific type of clustering, if present, should improve our understanding of the transmission and control of salmonellosis in the field.