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中国对虾(Litopenaeus vannamei)的血蓝蛋白基因的克隆与表达分析

Innate immune responses and metabolic alterations of mud crab (Scylla paramamosain) in response to Vibrio parahaemolyticus infection.

机构信息

Guangdong Provincial Key Laboratory of Marine Biology, Shantou University, Shantou, 515063, China; Marine Biology Institute, Shantou University, Shantou, 515063, China; STU-UMT Joint Shellfish Research Laboratory, Shantou University, Shantou, 515063, China.

Key Laboratory of Marine Genetic Resources, Third Institute of Oceanography, Ministry of Natural Resources, Xiamen, 361005, China.

出版信息

Fish Shellfish Immunol. 2019 Apr;87:166-177. doi: 10.1016/j.fsi.2019.01.011. Epub 2019 Jan 9.

Abstract

Vibrio parahaemolyticus is one of the major pathogens caused diseases in cultured mud crab (Scylla paramamosain). Mud crabs lack an adaptive immune system, their defenses depend almost on innate immunity. Evaluation of the molecular responses of mud crabs to pathogens is essential for control of disease occurrence in farmed animals. In this study, the impacts of V. parahaemolyticus on immunity-related genes and metabolites in mud crabs of different groups (PG, SG and MG refer to controlled, survival and moribund groups, respectively) were investigated. Our results revealed that V. parahaemolyticus infection stimulated significant expressions of immune-related genes (prophenoloxidase, alpha 2-macroglobulin, lysosomal-associated membrane protein, Rab5, C-type lectin B and anti-lipopolysaccharide factor 5) in the MG within 72 h post-infection. The ATP content was significantly reduced in all tissues except muscle of moribund mud crabs. A total of 668 metabolites (including 190 down-regulated and 145 up-regulated) were identified and assigned to 77 pathways in both SG and MG. Metabolites involved in the saturated fatty acid are up-regulated, whereas unsaturated fatty acid and amino acid metabolisms are down-regulated in the immune system of mud crabs during the bacterial infection in MG. Furthermore, a reduction of hemocyte number and an increase of microbial abundance was found in MG. Our results demonstrated that V. parahaemolyticus induced death of mud crabs through reducing the metabolites associate with energy biosynthesis and innate immune system (i.e. proliferation of hemocyte and melanization), resulting in decrease of ATP in different tissues and failed to clearance of pathogens, respectively. The findings of this study provide a basic information of the responses of mud crab on bacterial infection, which is essential for prevention and control of diseases in mud crab aquaculture.

摘要

副溶血弧菌是导致养殖泥蟹(锯缘青蟹)发病的主要病原体之一。泥蟹缺乏适应性免疫系统,其防御几乎完全依赖于先天免疫系统。评估泥蟹对病原体的分子反应对于控制养殖动物疾病的发生至关重要。在这项研究中,研究了副溶血弧菌对不同组(PG、SG 和 MG 分别指对照组、存活组和濒死组)泥蟹免疫相关基因和代谢物的影响。结果表明,副溶血弧菌感染在感染后 72 小时内刺激濒死组泥蟹中显著表达免疫相关基因(酚氧化酶原、α2-巨球蛋白、溶酶体相关膜蛋白、Rab5、C 型凝集素 B 和抗脂多糖因子 5)。除肌肉组织外,濒死组泥蟹所有组织中的 ATP 含量均显著降低。在 SG 和 MG 中鉴定出 668 种代谢物(包括 190 种下调和 145 种上调),并分配到 77 条途径中。在 MG 中,感染细菌期间,泥蟹免疫系统中的饱和脂肪酸代谢物上调,而不饱和脂肪酸和氨基酸代谢物下调。此外,在 MG 中还发现了血细胞数量减少和微生物丰度增加。研究结果表明,副溶血弧菌通过降低与能量生物合成和先天免疫系统相关的代谢物(即血细胞增殖和黑化)诱导泥蟹死亡,导致不同组织中 ATP 减少,无法清除病原体。本研究结果为泥蟹对细菌感染的反应提供了基础信息,这对于泥蟹养殖疾病的预防和控制至关重要。

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