Department of Fish Health and Environment, College of Fisheries, Central Agricultural University, Lembucherra, Post Box No. 60, Agartala 799 001, Tripura, India.
Fish Shellfish Immunol. 2012 Feb;32(2):353-9. doi: 10.1016/j.fsi.2011.11.030. Epub 2011 Dec 6.
Three different antigenic preparations from the epizootic ulcerative syndrome (EUS) pathogen Aphanomyces invadans were evaluated as vaccine candidate in catla (Catla catla). Anti-catla enzyme immunoconjugate was prepared after isolating catla immunoglobulin and raising hyperimmune sera against it, in rabbit. Three antigens namely, fungal extract (FE), fungal extract mixed with Freund's incomplete adjuvant (FIA) in a 1:1 (v/v) ratio (FE + A) and extra cellular product (ECP) were prepared and three groups of catla were vaccinated intramuscularly with all these antigens (200 μg/fish). Different cellular and humoral immune responses were measured for the entire vaccinated and control group on 0th, 5th, 15th and 25th day post vaccination. Thirty days after the vaccination, the fish were challenged with an A. invadans zoospore dose of 1 × 10(5) ml(-1) and mortality and relative percent of survival (RPS) were recorded. Study of cellular immunological parameters including antigen-specific leukocyte proliferation, antigen-specific nitric oxide production and superoxide anion production showed significantly higher (p < 0.05) values, in general, on 5th and 15th day post vaccination than the 0th day. Among all the antigenic groups, FE + A showed most significant response compared to the other groups. Among the humoral immune responses, lysozyme activity showed almost similar trend like cellular parameters. Anti-Aphanomyces antibody production was measured by enzyme-linked immunosorbent assay (ELISA) and it was increased with increasing days of vaccination in all the vaccinated groups with the highest observed on 25th day. Among the antigens, FE + A showed the highest antibody production following vaccination. The result of the homologous pathogen challenge study showed reduction in mortality in all the vaccinated groups. However, this reduction was not statistically significant (p > 0.05). Increased immune responses and protection have important implications with regard to the control of EUS by vaccination.
三种不同的抗原制剂，从流行溃疡综合征（EUS）病原体根肿菌（Aphanomyces invadans）被评估为候选疫苗在鲤鱼（Catla catla）。抗鲤鱼酶免疫结合物在兔中分离鲤鱼免疫球蛋白并针对其产生高免血清后制备。三种抗原，即真菌提取物（FE）、真菌提取物与弗氏不完全佐剂（FIA）以 1:1（v/v）的比例混合（FE+A）和细胞外产物（ECP），三组鲤鱼肌肉内接种所有这些抗原（200μg/fish）。在整个疫苗接种组和对照组的 0 天、5 天、15 天和 25 天，测量了不同的细胞和体液免疫反应。在接种后 30 天，用 1×10（5）ml（-1）的根肿菌游动孢子剂量对鱼进行攻毒，记录死亡率和相对存活率（RPS）。研究细胞免疫参数，包括抗原特异性白细胞增殖、抗原特异性一氧化氮产生和超氧阴离子产生，一般在接种后第 5 天和第 15 天比第 0 天显著升高（p<0.05）。在所有抗原组中，FE+A 与其他组相比表现出最显著的反应。在体液免疫反应中，溶菌酶活性与细胞参数几乎呈相似趋势。通过酶联免疫吸附试验（ELISA）测量抗根肿菌抗体的产生，在所有接种组中，随着接种天数的增加，抗体产生呈增加趋势，在第 25 天达到最高。在抗原中，FE+A 在接种后表现出最高的抗体产生。同源病原体攻毒研究的结果表明，所有接种组的死亡率均降低。然而，这一降低在统计学上并不显著（p>0.05）。免疫反应和保护的增加对通过疫苗接种控制 EUS 具有重要意义。
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