Screening of persistently infected cattle with bovine viral diarrhea virus on dairy farms by using milk tanker and bulk tank milk samples for viral RNA and viral-specific antibody detection.

The objective of this study was to provide a screening scheme of persistently infected (PI) cattle on dairy herds by combining reverse-transcription polymerase chain reaction (RT-PCR) to detect bovine viral diarrhea virus (BVDV) in milk tanker samples and commercial enzyme-linked immunosorbent assay to detect BVDV antibodies in bulk tank milk. We conducted a pilot survey and regional survey targeting all dairy farms in Ibaraki Prefecture by using milk tanker and bulk tank milk samples to screen PI cattle. Farms with positive samples underwent a follow-up test to identify PI cattle. In the pilot study, all virus-positive samples in bulk tank milk were included in the positive milk tanker samples. The RT-PCR assay successfully detected BVDV at dilutions of 1:1,600 by using two PI cows' milk. In the regional survey, 5 of 79 milk tanker samples were virus-positive. The virus was detected in three PI lactating cows and one PI calf on three farms. Antibody screening using bulk tank milk samples revealed 15 of 363 samples were positive, and 12 of 348 farms were BVDV antibody-positive. Follow-up tests on one farm identified three PI calves. Thus, eight PI cattle on five farms were identified in this study. In conclusion, combining BVDV detection using milk tanker samples and antibody detection using bulk tank milk is a feasible and economical method to efficiently screen PI cattle and confirm the PI-free status among dairy herds.

A novel genotype of avian infectious bronchitis virus isolated in Japan in 2009.

Avian infectious bronchitis viruses (IBVs) isolated from commercial layer flocks kept in Ibaraki Prefecture in 2009 were genetically and serologically characterized. Reverse transcription-PCR coupled with direct nucleotide sequencing and GenBank BLAST database analysis of the hypervariable region of the S1 subunit of the virus spike gene showed that these isolates are genetically very different from the previously known IBV genotypes in Japan. Furthermore, none of the antisera used in this study neutralized the index isolate (JP/Ibaraki/168-1/2009) in virus neutralization tests. These results suggest that the isolates are a novel IBV genotype in Japan (designated JP-IV).