Propagation of Chandipura virus in chick embryos.

Stocks of three Indian Chandipura virus (CHPV) isolates; one isolate from an adult febrile case in 1965 from Chandipura town, Maharashtra, and two isolates from two pediatric encephalitis cases from Andhra Pradesh, 2003 were inoculated in 10-day-old chick embryos by allantoic route. All three virus isolates replicated in chick embryos showing titre of log 10(12) to log 10(13) EID50. The results demonstrated that chick embryos are susceptible to CHPV and virus grows to high titres in this system. Therefore chick embryos can be used as an alternative host system for cultivation and isolation of CHPV as they are less expensive than laboratory animals and have several other advantages over cell cultures. Also this system can be used for the development of vaccine and diagnostic reagents.

Electron microscopy of buffalo green monkey kidney cells persistently infected with hepatitis A virus and immunolocalization of HAV antigens.

Studies were carried out to analyse the ultrastructural changes and the distribution of hepatitis A virus (HAV)/antigens at subcellular level in buffalo green monkey kidney (BGMK) cells persistently infected with HM-175 strain of HAV. HAV infected BGMK cells showed distinct abnormalities in the endoplasmic reticulum and cytoplasmic membrane as compared to uninfected cells. The abnormalities were characterized by wavy arrays, structures like myelin, annulate lamellae, cytoplasmic inclusion bodies and vesicles. The wavy arrays within the cytoplasm of the host cells appeared to represent degenerating membranes. A complex myelin like body was found in close association with a group of virus like particles. Annulate lamellae like structures involving single paired membrane were detected infrequently whereas the cytoplasmic vesicles were numerous in these cells. An indirect immunogold technique was utilized to localize the HAV antigenin infected cells. A high density immunogold label for HIV like particles was predominantly detected in cytoplasmic vesicles. These results suggest a strong association of membrane substructure in vesicle forms with the compartmentalized replication of HAV within persistently infected host cells.

Establishment of a cell line from Indian bonnet monkey (Macaca radiata) kidney and its characterization.

A cell line (MRK-90) has been established from a kidney tissue of a macaque monkey (Macaca radiata) of India. The cells are in 150th passage and have been characterized for morphology, chromosome number, isoenzyme patterns (LDH and MDH) and virus susceptibility. These studies indicate that the cells are epithelial like, heteroploid (2n = 65) and grow easily on glass surface/plastic surface without any difficulty. The cells are susceptible to a broad spectrum of viruses.