title: Development of an in vitro assay based on humoral immunity for quality control of oil-adjuvant Pseudotuberculosis vaccine in Yellowtail Seriola quinqueradiata.
abstract: Photobacterium damselae subsp. piscicida is an infectious pathogen that causes Pseudotuberculosis in Yellowtail fish. In Japan, several oil-adjuvant vaccines for Pseudotuberculosis have been approved for control of infectious diseases in aquaculture. Before distribution of an approved fish vaccine, an artificial challenge test for quality control is performed by the manufacturer and National Veterinary Assay Laboratory under Pharmaceutical Law of Japan to confirm potency. In this study, artificial challenge tests with a range of five diluted or undiluted approved vaccines was performed to determine the relationship between antigen levels and vaccine efficacy. Immunization of fish with the undiluted vaccine prevented Pseudotuberculosis. Results of artificial challenge tests demonstrated vaccine efficiency was dose dependent. Agglutination assays using immune sera were performed to determine agglutination titers, which were also dose dependent. These results suggest a link between survival rate in the artificial challenge tests and agglutination titers. Western blotting analysis identified a specific protein approximately 37 kDa in size in vaccinated fish. We confirmed antibodies were produced in vaccinated fish by immunoreactions with the approved vaccine. An agglutination assay based on humoral immunoreactions would be a useful alternative to the artificial challenge test for quality control of vaccines for aquaculture.
authors: National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, 1-15-1 Tokura, Kokubunji, Tokyo 185-8511, Japan. Electronic address: humiya_hirano@nval.maff.go.jp.
authors: Fumiya
authors: F
authors: Hirano
authors: National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, 1-15-1 Tokura, Kokubunji, Tokyo 185-8511, Japan.
authors: Saiki
authors: S
authors: Imamura
authors: National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, 1-15-1 Tokura, Kokubunji, Tokyo 185-8511, Japan.
authors: Nao
authors: N
authors: Nakajima
authors: National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, 1-15-1 Tokura, Kokubunji, Tokyo 185-8511, Japan.
authors: Kinya
authors: K
authors: Yamamoto
authors: National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, 1-15-1 Tokura, Kokubunji, Tokyo 185-8511, Japan.
authors: Mariko
authors: M
authors: Uchiyama
authors: National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, 1-15-1 Tokura, Kokubunji, Tokyo 185-8511, Japan.
authors: Hidetaka
authors: H
authors: Nagai
authors: National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, 1-15-1 Tokura, Kokubunji, Tokyo 185-8511, Japan.
authors: Mayumi
authors: M
authors: Kijima
mesh_terms: D000818:Animals; D000917:Antibody Formation; D001428:Bacterial Vaccines / Q000276:immunology* / Q000592:standards; D005393:Fish Diseases / Q000276:immunology / Q000517:prevention & control; D007106:Immune Sera; D066298:In Vitro Techniques; D010776:Photobacterium / Q000276:immunology*; D011786:Quality Control
chemical_list: D001428:Bacterial Vaccines; D007106:Immune Sera
keywords: Agglutination assay; NVAL; National Veterinary Assay Laboratory; PVDF; Photobacterium damselae subsp. Piscicida; Quality control; Seriola quinqueradiata; Vaccine; i.p.; intraperitoneally; polyvinylidene difluoride