Laboratory of Genome Science, Tokyo University of Marine Science and Technology, Tokyo, Japan; Department of Fisheries, Kasetklang Chatuchak, Bangkok, 10900, Thailand.
Songkhla Aquatic Animal Health Research Center, Department of Fisheries, Songkhla, Thailand.
Fish Shellfish Immunol. 2019 Jan;84:178-188. doi: 10.1016/j.fsi.2018.10.005. Epub 2018 Oct 4.
Acute hepatopancreatic necrosis disease (AHPND) is a shrimp farming disease, caused by the pathogenic Vibrio parahaemolyticus carrying a plasmid encoding Vp_PirAB-like toxins. Formalin-killed cells of V. parahaemolyticus AHPND-causing strain D6 (FKC-VpD6) were used to select Vp_PirAB-like toxin-resistant Litopenaeus vannamei by oral administration. Stomach and hepatopancreas tissues of shrimps that survived for one week were subjected to RNA sequencing. Differentially expressed genes (DEGs) between surviving shrimp, AHPND-infected shrimp, and normal shrimp were identified. The expressions of 10 DEGs were validated by qPCR. Only one gene (a gene homologous to L. vannamei anti-lipopolysaccharide factor AV-R isoform (LvALF AV-R)) was expressed significantly more strongly in the hepatopancreas of surviving shrimp than in the other groups. Significantly higher expression of LvALF AV-R was also observed in shrimp that survived two other trials of FKC-VpD6 selection. Recombinant ALF AV-R bound to LPS, PGN, Gram-negative bacteria, and some Gram-positive bacteria in ELISAs. ALF AV-R recombinant protein did not interact with native Vp_PirAB-like toxin in an ELISA or a Far-Western blot. For L. vannamei orally fed ALF AV-R protein for 3 days, the survival rate following challenge with VpD6-immersion was not significantly different from that of shrimp fed two control diets. These results suggest that LvALF AV-R expression was induced in the hepatopancreas of shrimp in response to the presence of Vp_PirAB-like toxin, although other factors might also be involved in the resistance mechanism.
急性肝胰腺坏死病（AHPND）是一种虾类养殖疾病，由携带编码 Vp_PirAB 样毒素质粒的致病性副溶血弧菌引起。采用携带编码 Vp_PirAB 样毒素质粒的副溶血弧菌 AHPND 致病株 D6（FKC-VpD6）福尔马林灭活细胞经口投喂的方法，对凡纳滨对虾进行 Vp_PirAB 样毒素抗性选择。存活 1 周的凡纳滨对虾的胃和肝胰腺组织进行 RNA 测序。鉴定存活虾、感染 AHPND 虾和正常虾之间的差异表达基因（DEGs）。通过 qPCR 验证了 10 个 DEGs 的表达。仅发现一个基因（与凡纳滨对虾抗脂多糖因子 AV-R 同工型（LvALF AV-R）同源的基因）在存活虾的肝胰腺中的表达显著高于其他两组。在另外两次 FKC-VpD6 选择试验中存活的虾中也观察到 LvALF AV-R 的表达显著升高。LvALF AV-R 在 ELISA 中与 LPS、PGN、革兰氏阴性菌和一些革兰氏阳性菌结合。在 ELISA 或 Far-Western blot 中，重组 ALF AV-R 蛋白与天然 Vp_PirAB 样毒素均无相互作用。连续 3 天经口投喂 ALF AV-R 蛋白的凡纳滨对虾，经 VpD6 浸泡攻毒后的存活率与投喂两种对照饲料的虾无显著差异。这些结果表明，LvALF AV-R 表达在虾的肝胰腺中被诱导，尽管其他因素也可能参与了抗性机制。
