Genetic analysis of the S1 gene of 4/91 type infectious bronchitis virus isolated in Japan.

S1 gene sequences for infectious bronchitis virus (IBV) strains of the 4/91 genotype (commonly called 793B) isolated from field outbreaks in Japan were analyzed to ascertain the relationship to 4/91 vaccine strain. Three field isolates (JP/Wakayama/2003, JP/Iwate/2005 and JP/Saitama/2006) from flocks not immunized with a 4/91 type live IBV vaccine and one isolate (JP/Wakayama-2/2004) from a flock immunized with a 4/91 type live vaccine were examined. The amino acid identities among JP/Wakayama/2003, JP/Iwate/2005 and JP/Saitama/2006 were about 98%, whereas the identities to the 4/91 vaccine strain and JP/Wakayama-2/2004 were about 90%. Three of the field isolates, JP/Wakayama/2003, JP/Iwate/2005 and JP/Saitama/2006, were classified into a cluster closely related to French and Spanish isolates, but different from the cluster including the vaccine and JP/Wakayama-2/2004. These results indicate that JP/Wakayama/2003, JP/Iwate/2005 and JP/Saitama/2006 were derived from foreign field isolates, but not from the vaccine strain. On the other hand, the S1 gene of JP/Wakayama-2/2004 revealed high sequence similarity with that of the 4/91 vaccine strain and appeared to be a vaccine-like virus derived from a vaccine. The field isolates of 4/91 genotype IBV could be distinguished from other genotypes by using the BalI and Pst I enzymes in addition to the polymerase chain reaction (PCR) -restriction fragment length polymorphism (RFLP) methods of Mase et al. [16] using Hae II and EcoR I enzymes. Furthermore, the 4/91 vaccine strain and vaccine-like isolate (JP/Wakayama-2/2004) could be differentiated from the other field isolates by Bgl II digestion. This method, therefore, would assist in identification of field isolates of the 4/91 genotype as outbreaks of IBV in vaccinated flocks.

Isolation of 4/91 type of infectious bronchitis virus as a new variant in Japan and efficacy of vaccination against 4/91 type field isolate.

Among field isolates of infectious bronchitis virus (IBV) from recent outbreaks, some isolates that were classified into the 4/91 genotype, by analysis of the S1 gene, have been confirmed to be a new variant in Japan. To elucidate the characteristics of these isolates, pathogenicity in chicks and the efficacy of vaccines against this new variant were examined. Severe respiratory symptoms were observed in 4-day-old specific pathogen free chicks inoculated with a 4/91 genotype isolate, either JP/Wakayama/2003 or JP/ Iwate/2005; body weights 3 wk after inoculation were significantly lower than those of chicks inoculated with a 4/91 vaccine strain. These 4/91 isolates were neutralized with serum from birds immunized with 4/91 vaccine. In a challenge-protection test, five groups of chicks were immunized with C78, TM-86w, H120, Kita-1, or 4/91 vaccines and then challenged with JP/Iwate/2005 4 wk after vaccination. A protective effect in the 4/91 and TM-86w vaccine groups was indicated by evaluation of the ciliostasis score of the trachea, the respiratory symptom score, and virus isolation from trachea swab samples after challenge. The results of this study suggested that the 4/91 type of IBV, which is virulent to chicks when compared to vaccine strains, has emerged as a new variant in Japan, and vaccines containing the 4/91 strain or the TM-86w strain could be effective for this variant.