Fecal extracts from 12 subjects in outbreaks of oyster-associated nonbacterial gastroenteritis were inoculated with BS-C-1 cells for isolation of the causative viruses. Cytopathic agents were isolated from 3 patients. No cross-neutralizing reactions were observed between the isolates and prototypes of human enteroviruses. The isolates were approximately 30 nm in diameter and had a distinct ultrastructure resembling that of astroviruses. Four polypeptide bands with molecular sizes of 42, 28, 27, and 22 kDa were seen on SDS-PAGE analyses. Seroconversion against the isolate was observed in 18 (31.6%) of 57 patients involved in five of seven outbreaks examined by neutralization test. A protein band characteristically reactive with the paired serum samples was detectable at 42 kDa by immunoblot assay. These results suggested that some small round viruses resembling astroviruses might show cytopathic effect in BS-C-1 cells and may be associated with an oyster-related gastroenteritis.

Viral particles morphologically resembling animals caliciviruses in the faeces of a patient with acute gastroenteritis were purified, radiolabeled with [125I], and analyzed by SDS-PAGE. A single major structural protein with a mol. mass 62,000 daltons was identified by immunoprecipitation technique. The finding is consistent with human calicivirus-like particles associated with gastroenteritis being a member of the family Caliciviridae.

Human adenovirus type 40 (Ad40) cannot propagate in conventional established human cell lines such as KB or HeLa cells. However, it has been shown that Ad40 DNA replicates in KB18 cells which express Ad2 E1B genes, suggesting that Ad40 is defective in the E1B gene function in KB or HeLa cells. We show here that Ad40 can propagate and produce plaques on A549 cells which do not contain Ad E1B genes. Our experiments show that the levels of replication of Ad40 DNA and production of infectious Ad40 virus in A549 cells are the same as or higher than those in 293 or KB18 cells. Dot blot analysis shows that the levels of Ad40 E1A and E1B mRNAs expressed in A549 cells at early to intermediate times postinfection are at least 10-fold higher than those in KB or KB18 cells. Northern (RNA) blot analysis shows that large E1B mRNA species (approximately 24S to 26S) are synthesized prior to the onset of DNA replication in A549 cells. No E1B mRNA species are synthesized in KB or KB18 cells at early times postinfection, and no differences in the expression of E1B mRNAs are seen between KB and KB18 cells. The experiment suggests that A549 cells have a cellular factor(s) which activates Ad40 E1B mRNA synthesis and that the E1B mRNA synthesis helps Ad40 propagation. In contrast, Ad40 can propagate in KB18 cells by using Ad2 E1B gene products that are constitutively expressed in this cell line. Furthermore, this result shows that Ad40 cannot propagate in KB cells because of the failure in the expression of E1B genes at early times postinfection.

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