Development of improved diagnostics and pathotyping methods for IBDV in Australia

Commonwealth Scientific and Industrial Research Organisation (CSIRO)

  • Project code: PRO-015885

  • Project stage: Current

  • Project start date: Saturday, April 29, 2023

  • Project completion date: Wednesday, April 29, 2026

  • National Priority: CME-Priority 2-Enhancing chicken biosecurity, health, and welfare

Summary

IBDV is continuing to evolve around the world. Since the emergence of very virulent IBDV (vvIBDV) and antigenic variants in the mid 80’s, genetically distinct genotypes of IBDV have continued to be identified in different countries. To date, 8 genogroups of IBDV (A1 to A8) have been identified worldwide, with Australian strains comprising genogroups A7 and A8. It is therefore crucial that we continue to monitor strains circulating worldwide to ensure that the current tests being used here in Australia are able to rapidly detect all exotic strains of IBDV and differentiate them from endemic IBDV strains. Currently, a single qPCR test is used to detect any circulating IBDV strains in Australia, followed by nucleotide sequencing of the viral VP2 gene to determine the identity of the IBDV isolate. This need to sequence samples to determine the type of IBDV involved limits both the sensitivity and number of isolates that can be sequenced on any given day (currently 48 samples per 8 hr work period). Hence, we aim to develop new qPCR tests which can differentiate all exotic IBDV strains (genogroups A1 to A6) from endemic Australian strains (genogroups A7 and A8) to improve the sensitivity and turnaround time in identifying exotic strains of IBDV. To assist with the design of primers and probes required for qPCR tests, whole genome sequencing of Australian IBDV will also be undertaken. Once developed, these qPCR assays will greatly enhance our ability to quickly differentiate exotic strains from endemic strains in the event of an incursion, thus improving outbreak response times and the chances of eradication.

In Australia, we have been pathotyping field isolates of IBDV using the protocol based on OIE guidelines, which recommends the use of 3-6 week old chickens and a virus dose of 105 50% egg infectious dose (EID50). This high dose was originally recommended to define what constitutes a “vvIBDV phenotype”. Strains producing greater than 50% mortality were classified as vvIBDV. The protocol was not designed to depict the disease that is typically observed in the field. Similarly, it was not designed to detect any pathotypic differences amongst strains that do not induce mortalities in SPF birds. This protocol was chosen to confirm that Australian strains not cause any mortalities in SPF chickens using a dose of 105 EID50. Unfortunately, due to the high dose of virus given, it is often difficult to detect differences amongst the Australian strains which are statistically significant. Variants typically produce slightly greater histopathological changes in the bursa, with average lymphocyte depletion scores reaching the maximum score of 4 (on a scale of 1-4) compared to classical strains which average scores closer to 3. Recently however, we have been observing more classical strains reaching lymphocyte depletion scores approaching 4. Hence our scale of comparison has effectively reached its peak. The significance of these high lymphocyte depletion scores observed in the bursa in terms of flock performance is uncertain as we have never tested the immunosuppression of any Australian strains.

Although Australian strains induce no mortalities in SPF chickens using the protocol recommend by the OIE above, Australian variants do typically induce mild to moderate clinical signs in chickens. We believe the appearance of clinical signs is dose dependent. We recently tested a Victorian variant at a lower dose of 104 EID5

Program

Chicken Meat

Research Organisation

Commonwealth Scientific and Industrial Research Organisation (CSIRO)