Effects of Aeromonas hydrophila Stimulation on Immunity of Chinese Mitten Crab Eriocheir sinensis
CHENG Chao1, XIAO Min2, LI Jing1, MAO Zhenfang1, ZHANG Cuizhen1, CHEN Zhihe1, ZHU Minjie1, JIAN Shaoqing1,3, ZHAO Daxian1,3
1. School of Life Sciences, Nanchang University, Nanchang 330031, China; 2. Ji'an City Jinggang Mountains Agricalture Science and Technology Park in Jiangxi Province, Ji'an 343000, China; 3. Jiangxi Provincal Key Laboratory of Aquatic Animal Resources and Utilization, Nanchang 330031, China
Abstract：In this study, the influence of Aeromonas hydrophila stimulation on immune tissue structure and gene expression of Chinese mitten crab Eriocheir sinensis was analyzed by conventional paraffin section, hematoxylin-eosin staining (HE staining) and real-time fluorescence quantitative PCR.The results showed that, with the bacterial stimulation, the gill filaments thickness and the number of blood cells of Chinese mitten crab were decreased and then increased, the lumen area, structure and cell number of hepatic small tubules were changed significantly compared with the control group.The expression levels of lipopolysaccharide and beta-1, 3-glucan binding protein (LGBP) were decreased significantly at 6 h (P<0.05), but increased significantly at 12 h (P<0.05). Tube gene expression level was decreased significantly at 3 h and 6 h (P<0.05), and increased significantly at 12 h (P<0.05). Dorsal expression levels were increased significantly at 3 h (P<0.05), The expression level of integrin was significantly increased at 3 h and 6 h after infection (P<0.05), and was significantly increased at 12 h (P<0.01). The expressions of prophenoloxidase activating factor (PPAF), peroxinectin (PXN) and glutathione S-transferase (GST) were found to be decreased gradually within 12 h after infection. The expression levels of glutathione peroxidase (GPx) reached the lowest value at 6 h after infection, and then was upregulated. The expression level of superoxide dismutase (SOD) was significantly decreased at 6 h after infection (P<0.05). In conclusion, Aeromonas hydrophila stimulation significantly changed the structures of the gill and hepatopancreas and the expression levels of immune-related genes in Chinese mitten crab.
[1]农业部渔业渔政局. 2016中国渔业统计年鉴[M].北京:中国农业出版社,2016. [2]姜艳丽,李丽丽. 嗜水气单胞菌的研究进展[J]. 黑龙江农业科学,2015(2):142-146. [3]郑杰夫,张一柳,钟蕾. 中华鳖源嗜水气单胞菌的研究进展[J]. 水产科学,2016,35(2):191-198. [4]余雪明,金春光,章丹阳,等. 致病性气单胞菌在贝(甲)壳类海产品中的检出[J]. 中国卫生检验杂志,2009,19(4):928-930. [5]衣启麟. 养殖虾蟹疫病免疫防控的初步研究[D]. 北京:中国科学院大学,2014. [6]Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CT method[J]. Methods,2001,25(4):402-408. [7]Allan B J, Stevenson R M W. Extracellular virulence factors of Aeromonas hydrophila, in fish infections[J]. Canadian Journal of Microbiology,1981,27(10):1114-1122. [8]沈锦玉,钱冬,刘问,等. 养殖青虾“红鳃病”病原的研究[J]. 浙江海洋学院学报:自然科学版,2000,19(3):222-224. [9]Yoganandhan K, Leartvibhas M, Sriwongpuk S, et al. White tail disease of the giant freshwater prawn Macrobrachium rosenbergii in Thailand[J]. Diseases of Aquatic Organisms,2006,69(2/3):255-258. [10]沈锦玉,尹文林,钱冬,等. 中华绒螯蟹“腹水病”及“抖抖病”并发病病原的研究[J]. 中国水产科学,2000,7(3):89-92. [11]汤显春,薛翠峰,高智慧. 沼泽绿牛蛙致病细菌的研究[J]. 水生态学杂志,2000,20(1):27-28. [12]Rigney M M, Zilinsky J W, Rouf M A. Pathogenicity of Aeromonas hydrophila in red-leg disease in frogs[J]. Current Microbiology,1978,1(3):175-179. [13]Hazen T C, Raker M L, Esch G W, et al. Ultrastruct of red-sore lesions on largemouth bass (Micropterus salmoides): associattion of the ciliate Epistylis sp. and the bacterium Aeromonas hydrophila[J]. Journal of Protozoology,2010,25(3):351-355. [14]陈怀青,陆承平. 家养鲤科鱼暴发性传染病的病原研究[J]. 南京农业大学学报,1991,14(4):87-91. [15]Rahman M M, Chowdhury M B R. Incidence of ulcer disease in African catfish (Clarias gariepinus Burchell) and trial for its chemotherapy[J]. Bangladesh Journal of Fisheries Research,1999,3(2):193-200. [16]Bootsma R. Hydrocephalus and red-disease in pike fry Esox lucius L.[J]. Journal of Fish Biology,1971,3(4):417-419. [17]赵艳民. 水体Hg2+对中华绒螯蟹毒性作用研究[D]. 天津:南开大学,2009. [18]顾志峰,王文,杜开和. 中华绒螯蟹鳃的超微结构观察[J]. 电子显微学报,2000,19(3):141-142. [19]卢敬让,赖伟. 镉对中华绒螯蟹鳃组织及其亚显微结构的影响[J]. 海洋与湖沼,1991,22(6):566-570. [20]张瑶,潘连德. 蟹类肝胰腺病症的病原和病理学研究进展[J]. 水产科技情报,2012,39(3):136-139. [21]张瑶. 中华绒螯蟹肝胰腺病症的病理分析[D]. 上海:上海海洋大学,2013. [22]洪美玲,陈立侨,顾顺樟,等. 氨氮胁迫对中华绒螯蟹免疫指标及肝胰腺组织结构的影响[J]. 中国水产科学,2007,14(3):412-418. [23]杨志彪,赵云龙,杨健. 水体铜对中华绒螯蟹鳃和肝胰腺显微结构及金属硫蛋白含量的影响[G]//中国水产学会青年学术年会摘要集.北京:中国水产学会,2004. [24]于爱清. 虾蟹类免疫相关基因的研究[D]. 上海:华东师范大学,2014. [25]赵大显. 中华绒螯蟹表达序列标签(EST)分析及免疫相关基因的克隆、表达模式研究[D].上海:华东师范大学,2009. [26]Iwanaga S, Lee B L. Recent advances in the innate immunity of invertebrate animals[J]. Journal of Biochemistry and Molecular Biology,2005,38(2):128-150. [27]Zhao D X, Chen L Q, Qin C J, et al. Molecular cloning and characterization of the lipopolysaccharide and β-1, 3-glucan binding protein in Chinese mitten crab (Eriocheir sinensis)[J]. Comparative Biochemistry and Physiology B,2009,154(1):17-24. [28]Yu A Q, Jin X K, Wu M H, et al. Identification and characterization of tube in the Chinese mitten crab Eriocheir sinensis[J].Gene,2014,541(1):41-50. [29]Yu A Q, Jin X K, Li S, et al. Molecular cloning and expression analysis of a dorsal homologue from Eriocheir sinensis[J]. Developmental and Comparative Immunology,2013,41(4):723-727. [30]Huang Y, Zhao L L, Feng J L, et al. A novel integrin function in innate immunity from Chinese mitten crab (Eriocheir sinensis)[J]. Developmental & Comparative Immunology,2015,52(2):155-165. [31]于杰伦. 日本沼虾酚氧化酶原激活因子基因克隆及其表达分析[D].泰安:山东农业大学,2016. [32]Kobayashi M, Johansson M W, Söderhäll K. The 76 kD cell-adhesion factor from crayfish haemocytes promotes encapsulation in vitro[J]. Cell and Tissue Research,1990,260(1):13-18. [33]Lv S, Lu B, Xu J, et al. Immune response of peroxinectin of Chinese mitten crab Eriocheir sinensis to exterior stimulation[J].Developmental and Comparative Immunology,2015,51(1):56-64. [34]Zhao D X, Chen L Q, Qin C J, et al.A delta-class glutathione transferase from the Chinese mitten crab Eriocheir sinensis:cDNA cloning, characterization and mRNA expression[J]. Fish & Shellfish Immunology,2010,29(4):698-703. [35]Meng Q, Jie D U, Wei Y, et al. An extracellular copper/zinc superoxide dismutase (Cu Zn SOD) from Chinese mitten crab, Eriocheir sinensis and its relationship with Spiroplasma eriocheiris[J]. Aquaculture,2011,320(1):56-61. [36]董亮,何永志,王远亮,等. 超氧化物歧化酶(SOD)的应用研究进展[J]. 中国农业科技导报,2013,15(5):53-58. [37]Mu C K, Zhao J M, Wang L L, et al. Molecular cloning and characterization of peroxiredoxin 6 from Chinese mitten crab Eriocheir sinensis[J]. Fish & Shellfish Immunology,2008,26(6):821-827. [38]Zhao D X, Chen L Q, Qin J G, et al. Molecular characterization of a cytosolic manganese superoxide dismutase from the Chinese mitten crab Eriocheir sinensis [J]. Genetics and Molecular Research,2014,13(4):9429-9442.
2020, Vol. 39 
