Development of an enzyme-linked immunosorbent assay for the serological detection of exposure of poultry in New Zealand to Erysipelothrix rhusiopathiae and their serological response to vaccination

Abstract

AIMS: To modify and validate an existing swine erysipelas ELISA for use with poultry serum and to assess the safety of a swine erysipelas vaccine for use in New Zealand layer birds.

METHODS: An existing swine erysipelas ELISA was modified for use in domestic poultry and was validated using sera from birds injected with either 2 mL of a commercially available killed swine erysipelas vaccine (low-dose; n=12 birds), 4 mL of vaccine (high-dose; n=11 birds), or 2 mL saline (control; n=11 birds) on Day 0 and again on Day 21. Blood samples were collected on Days 0, 21, 42, and 63, and safety of the vaccine for use in layer birds was determined by assessing cloacal temperature and injection site reactions in birds at 0, 4, 24, 48, 72 and 96 h post-vaccination.

RESULTS: The ELISA that was developed had a diagnostic sensitivity and specificity of 93% and 98%, respectively, after being optimised for a positive cut-off at an optical density (OD) ≥1.50 read at 450-nm wavelength. OD readings were higher on Days 21, 42, and 63 than Day 0 in both the low-dose and high-dose groups (p<0.05), and differed amongst the three groups on Days 21, 42, and 63 (p<0.05), suggesting that vaccination using either dose induced detectable levels of antibody, even after a single dose. In addition, the high-dose protocol induced higher levels of antibody production than the low-dose protocol. No local or systemic reactions to the vaccine were observed and cloacal temperatures remained in the normal biological range after vaccination.

CONCLUSIONS: The ELISA that was developed had satisfactory diagnostic performance characteristics and the vaccine appeared to be safe for use in layer birds. However, the study design did not permit an assessment of the vaccine's efficacy to protect birds from clinical erysipelas.

CLINICAL RELEVANCE: A diagnostic ELISA has been developed for determining the exposure of layer birds to E. rhusiopathiae. The test will be useful for monitoring flock-level erysipelas, response to vaccination, and in epidemiological studies designed to identify risk factors for exposure to the disease.

Key Words:

• Erysipelas
• ELISA
• poultry
• validation
• vaccine safety

Acknowledgements

The authors wish to acknowledge the assistance of Kathryn Bonistalli in modification and optimisation of the ELISA. Additionally, support and advice of colleagues in the Massey University Molecular Epidemiology and Public Health Laboratory are appreciated. Advice and guidance from Dukhpati Rao in the Institute of Veterinary, Animal, and Biomedical Sciences is acknowledged. We thank Dr. Chris Morrow, Bioproperties Pty Ltd, Australia for his support. Furthermore, the assistance of Don Thomas and staff at the Massey University Poultry Farm in caring for the birds during the study is appreciated. We are grateful for the financial support of the Egg Producers Charitable Trust.

Notes

*Non-peer-reviewed