Isolation of BCoV was performed in monolayers of Hm. Lu-1 cells, using 20 fecal samples from clinical cases of diarrhea in calves. Samples were positive for BCoV by means of hemagglutination/hemagglutination inhibition (HA/HI). Up to the fifth serial passage, 13 of these isolates presented syncytial cytopathic effect, similar to Kakegawa standard strain. When isolates were submitted to seroneutralization with gammaglobulin anti-bovine coronavirus, 8 of them were considered to be positive, once cytopathic effect was neutralized. After titration and seroneutralization in microplates, only three of them were confirmed as positive. The lineage Hm. Lu-1 showed higher permissivity to BCoV isolation, but the low intensity of viral replication demonstrated that new methodologies should be developed for this purpose and then submitted to confirmation of isolation by means of seroneutralization.
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