Corado Maria Vitória Lobo, Pereira Elcimara Cardoso, Botinelly Taísa Freitas, Glória Juliane Corrêa, Sousa Rafael Luckwu de, Corado André de Lima Guerra, Balieiro Antônio Alcirley da Silva, Mariúba Luís André Morais, Gallani Silvia Umeda, Tavares Guilherme Campos
Universidade Nilton Lins, Manaus 69058-030, AM, Brazil.
Instituto Leônidas e Maria Deane, Fiocruz Amazônia, Manaus 69057-070, AM, Brazil.
Animals (Basel). 2025 Mar 18;15(6):869. doi: 10.3390/ani15060869.
Infections with pose risks to the production of . Since no therapeutic treatment exists for this disease in this aquatic host, prophylaxis through vaccination is one method for disease control. The enzyme-linked immunosorbent assay (ELISA) is widely used in aquaculture to assess animal immunity and, mainly, to validate vaccination protocols. The method based on the use of immunoglobulin Y (IgY) is an alternative to mammalian antibodies that should be considered due to its advantages over antibodies produced by small mammals. This study aimed to standardize an indirect ELISA (iELISA) protocol to validate the immunization of against . For this, a hen and fish were immunized to obtain anti- and anti- antibodies, respectively, which were used to standardize the test. Fish were inoculated with bacterin + adjuvant, bacterin alone, adjuvant alone, or phosphate-buffered saline, subjected to blood sampling, and monitored for up to 60 days. The iELISA was composed of the inactivated bacteria in the coating step, anti- as the primary antibody, and anti- IgY as the secondary antibody. The results demonstrate that the highest antibody titers were found in the organisms immunized with bacterin + adjuvant and bacterin. The iELISA enabled the detection of antibodies produced by , which were mediated by IgY specificity.
感染 会对 的生产造成风险。由于在这种水生宿主中不存在针对该疾病的治疗方法，通过疫苗接种进行预防是疾病控制的一种方法。酶联免疫吸附测定（ELISA）在水产养殖中广泛用于评估动物免疫力，主要用于验证疫苗接种方案。基于免疫球蛋白Y（IgY）的方法是哺乳动物抗体的一种替代方法，由于其相对于小型哺乳动物产生的抗体具有优势，应予以考虑。本研究旨在标准化一种间接ELISA（iELISA）方案，以验证 针对 的免疫接种。为此，分别对一只母鸡和鱼进行免疫，以获得抗 和抗 抗体，用于标准化该试验。给鱼接种菌苗+佐剂、单独的菌苗、单独的佐剂或磷酸盐缓冲盐水，进行采血，并监测长达60天。iELISA在包被步骤中由灭活细菌组成，抗 作为一抗，抗 IgY作为二抗。结果表明，在用菌苗+佐剂和菌苗免疫的生物体中发现了最高的抗体滴度。iELISA能够检测由 产生的抗体，这些抗体由IgY特异性介导。
