Abstract：In order to explore the effect of inactivated infectious spleen and kidney necrosis virus (ISKNV) vaccine on the composition, structure and related functional genes of gut microbes in mandarin fish Siniperca chuatsi, the mandarin fish with body weight of (20.0±1.8) g was intraperitoneally injected with the inactivated ISKNV vaccine, and was injected with an equal volume of sterile saline in the control group. The fish in the control group and the immunized group were reared in the same pond separated by an enclosure for 28 days when 10 intestinal fecal samples were collected from each group, and sequenced by high-throughput sequencing to construct a library. A total of 9669 Unigenes with an average length of 543 bp were obtained by high-throughput sequencing. Alpha diversity analysis showed that Chao1 index was higher in the control group, and Shannon index and Simpson index were higher in the immunized group, indicating that the variety was decreased and the diversity and richness were shown to be increased in gut microbes of the immunized mandarin fish. Proteobacteria was the dominant phyla in the control group, with abundance of 72.23%, at the phylum level, with the relative abundance of 28.57% in Proteobacteria and 11.13% in Chlamydia in the immune group. At the genus level, Aeromonas was the dominant genera in the control group, with the relative abundance of 71.73%. In the immunized group, Chlamydia and Escherichia were the dominant genera, with the abundance of 11.57% and 8.7%, respectively. Compared with the control group, 9656 differentially genes were enriched in the immunized group, with 840 high abundance genes and 8816 low abundance genes. Differential genes were mainly enriched in the two-component system, ABC transporter, purine metabolism, quorum sensing, bacterial chemotaxis, flagellar assembly and bacterial secretion system, and the abundance of most genes in the pathway were decreased in the immunized group. Injection administration of inactivated ISKNV vaccine resulted in changes in intestinal microflora and functional genes, with decrease in proportions of Aeromonas hydrophila and A. veronii, and increase in proportions of intestinal beneficial bacteria such as Lactobacillus, Bacillus and Paenibacillus, which provides new insights into immune mechanism of vaccines.
[1] 吴淑勤,李新辉,潘厚军,等.鳜暴发性传染病病原研究[J].水产学报,1997,21(增刊):56-60. [2] 何建国,翁少萍,黄志坚,等.鳜暴发流行病病毒性病原研究[J].中山大学学报(自然科学版),1998,37(5):74-77. [3] WANG Y Q, LÜ L, WENG S P, et al. Molecular epidemiology and phylogenetic analysis of a marine fish infectious spleen and kidney necrosis virus-like (ISKNV-like) virus[J].Archives of Virology,2007,152(4):763-773. [4] DONG C F, XIONG X P, LUO Y W, et al. Efficacy of a formalin-killed cell vaccine against infectious spleen and kidney necrosis virus (ISKNV) and immunoproteomic analysis of its major immunogenic proteins[J].Veterinary Microbiology,2013,162(2/3/4):419-428. [5] FU X, LI N, LAI Y, et al. A novel fish cell line derived from the brain of Chinese perch Siniperca chuatsi:development and characterization[J].Journal of Fish Biology,2015,86(1):32-45. [6] 潘厚军,吴涉勤.鳜鱼疫苗预防鳜鱼病毒性暴发性传染病试验[J].水产科技,2000(6):18-21. [7] PÉREZ T, BALCÁZAR J L, RUIZ-ZARZUELA I, et al. Host-microbiota interactions within the fish intestinal ecosystem[J].Mucosal Immunology,2010,3(4):355-360. [8] LIN C S, CHANG C J, LU C C, et al. Impact of the gut microbiota, prebiotics, and probiotics on human health and disease[J].Biomedical Journal,2014,37(5):259-268. [9] CORNEJO-GRANADOS F, GALLARDO-BECERRA L, LEONARDO-REZA M, et al. A meta-analysis reveals the environmental and host factors shaping the structure and function of the shrimp microbiota[J].PeerJ,2018,6:e5382. [10] CORNEJO-GRANADOS F, LOPEZ-ZAVALA A A, GALLARDO-BECERRA L, et al. Microbiome of Pacific whiteleg shrimp reveals differential bacterial community composition between wild, aquacultured and AHPND/EMS outbreak conditions[J].Scientific Reports,2017,7:11783. [11] DUAN Y F, ZHANG Y, DONG H B, et al. .Effects of dietary poly-β-hydroxybutyrate (PHB) on microbiota composition and the mTOR signaling pathway in the intestines of Litopenaeus vannamei[J].Journal of Microbiology,2017,55(12):946-954. [12] LI X M, YAN Q Y, RINGØ E, et al. The influence of weight and gender on intestinal bacterial community of wild largemouth bronze gudgeon (Coreius guichenoti, 1874)[J].BMC Microbiology,2016,16(1):191. [13] LAUZON H L, GUDMUNDSDOTTIR S, PETURSDOTTIR S K, et al. Microbiota of Atlantic cod (Gadus morhua L.) rearing systems at pre-and posthatch stages and the effect of different treatments[J].Journal of Applied Microbiology,2010,109(5):1775-1789. [14] YANG H T, ZOU S S, ZHAI L J, et al. Pathogen invasion changes the intestinal microbiota composition and induces innate immune responses in the zebrafish intestine[J].Fish & Shellfish Immunology,2017,71:35-42. [15] THAISS C A, ZMORA N, LEVY M, et al. The microbiome and innate immunity[J].Nature,2016,535(7610):65-74. [16] SHARMA R, SCHUMACHER U, RONAASEN V, et al. Rat intestinal mucosal responses to a microbial flora and different diets[J].Gut,1995,36(2):209-214. [17] PETERSSON J, SCHREIBER O, HANSSON G C, et al. Importance and regulation of the colonic mucus barrier in a mouse model of colitis[J].American Journal of Physiology.Gastrointestinal and Liver Physiology,2011,300(2):G327-G333. [18] HAMADA H, HIROI T, NISHIYAMA Y, et al. Identification of multiple isolated lymphoid follicles on the antimesenteric wall of the mouse small intestine[J].The Journal of Immunology,2002,168(1):57-64. [19] BOUSKRA D, BRÉZILLON C, BÉRARD M, et al. Lymphoid tissue genesis induced by commensals through NOD1 regulates intestinal homeostasis[J].Nature,2008,456(7221):507-510. [20] IVANOV I I, ATARASHI K, MANEL N, et al. Induction of intestinal Th17 cells by segmented filamentous bacteria[J].Cell,2009,139(3):485-498. [21] CHU H, KHOSRAVI A, KUSUMAWARDHANI I P, et al. Gene-microbiota interactions contribute to the pathogenesis of inflammatory bowel disease[J].Science,2016,352(6289):1116-1120. [22] ATARASHI K, TANOUE T, SHIMA T, et al. Induction of colonic regulatory T cells by indigenous Clostridium species[J].Science,2011,331(6015):337-341. [23] ZHANG C X, WANG H Y, CHEN T X. Interactions between intestinal microflora/probiotics and the immune system[J].BioMed Research International,2019,2019:6764919. [24] BROWN E M, SADARANGANI M, FINLAY B B. The role of the immune system in governing host-microbe interactions in the intestine[J].Nature Immunology,2013,14(7):660-667. [25] RAWLS J F, SAMUEL B S, GORDON J I. Gnotobiotic zebrafish reveal evolutionarily conserved responses to the gut microbiota[J].Proceedings of the National Academy of Sciences of the United States of America,2004,101(13):4596-4601. [26] 王纯,倪加加,颜庆云,等.草鱼与团头鲂肠道菌群结构比较分析[J].水生生物学报,2014,38(5):868-875. [27] 祭仲石,管卫兵,苏孙国,等.鲢、鳙肠道微生物的研究[J].大连海洋大学学报,2014,29(1):22-26. [28] LIU H, GUO X W, GOONERATNE R, et al. The gut microbiome and degradation enzyme activity of wild freshwater fishes influenced by their trophic levels[J].Scientific Reports,2016,6:24340. [29] RUNGRASSAMEE W, KLANCHUI A, MAIBUNKAEW S, et al. Characterization of intestinal bacteria in wild and domesticated adult black tiger shrimp (Penaeus monodon)[J].PLoS One,2014,9(3):e91853. [30] 张涵,周涛,王岩.综合养殖池塘中三角帆蚌和鱼类肠道细菌的组成[J].水生生物学报,2013,37(5):824-835. [31] LI T T, LONG M, JI C, et al. Alterations of the gut microbiome of largemouth bronze gudgeon (Coreius guichenoti) suffering from furunculosis[J].Scientific Reports,2016,6:30606. [32] 张燕玉,韩卓然,孙敬锋,等.海藻希瓦氏菌感染对半滑舌鳎肠道菌群结构及相关功能基因表达的影响[J].南方农业学报,2019,50(10):2300-2307. [33] 张正,廖梅杰,李彬,等.两种疾病发生对养殖半滑舌鳎肠道菌群结构的影响分析[J].水产学报,2014,38(9):1565-1572. [34] 刘志刚,卢迈新,可小丽,等.尼罗罗非鱼肠道及养殖环境中菌群结构与链球菌病的相关性[J].水产学报,2018,42(10):1635-1647. [35] XIA J H, LIN G, FU G H, et al. The intestinal microbiome of fish under starvation[J].BMC Genomics,2014,15:266. [36] HE S X, WANG Q M, LI S N, et al. Antibiotic growth promoter olaquindox increases pathogen susceptibility in fish by inducing gut microbiota dysbiosis[J].Science China Life Sciences,2017,60(11):1260-1270. [37] CLARKE S F, MURPHY E F, O'SULLIVAN O, et al. Exercise and associated dietary extremes impact on gut microbial diversity[J].Gut,2014,63(12):1913-1920. [38] JOHNSON P T J, HARTSON R B, LARSON D J, et al. Diversity and disease:community structure drives parasite transmission and host fitness[J].Ecology Letters,2008,11(10):1017-1026. [39] ZHU J Y, DAI W F, QIU Q F, et al. Contrasting ecological processes and functional compositions between intestinal bacterial community in healthy and diseased shrimp[J].Microbial Ecology,2016,72(4):975-985. [40] MARKOWIAK P, ŚLIZ·EWSKA K. Effects of probiotics, prebiotics, and synbiotics on human health[J].Nutrients,2017,9(9):1021. [41] TOJO R, SUÁREZ A, CLEMENTE M G, et al. Intestinal microbiota in health and disease:role of bifidobacteria in gut homeostasis[J].World Journal of Gastroenterology,2014,20(41):15163-15176. [42] LIU C H, CHIU C H, WANG S W, et al.. Dietary administration of the probiotic, Bacillus subtilis E20, enhances the growth, innate immune responses, and disease resistance of the grouper, Epinephelus coioides[J].Fish & Shellfish Immunology,2012,33(4):699-706. [43] SON V M, CHANG C C, WU M C, et al. Dietary administration of the probiotic, Lactobacillus plantarum, enhanced the growth, innate immune responses, and disease resistance of the grouper Epinephelus coioides[J].Fish & Shellfish Immunology,2009,26(5):691-698. [44] CARNEVALI O, ZAMPONI M C, SULPIZIO R, et al. Administration of probiotic strain to improve sea bream wellness during development[J].Aquaculture International,2004,12(4/5):377-386. [45] BAGHERI T, HEDAYATI S A, YAVARI V, et al. Growth, survival and gut microbial load of rainbow trout (Onchorhynchus mykiss) fry given diet supplemented with probiotic during the two months of first feeding[J].Turkish Journal of Fisheries and Aquatic Sciences,2008,8(1):43-48. [46] ZOKAEIFAR H, BALCÁZAR J L, SAAD C R, et al. Effects of Bacillus subtilis on the growth performance, digestive enzymes, immune gene expression and disease resistance of white shrimp, Litopenaeus vannamei[J].Fish & Shellfish Immunology,2012,33(4):683-689. [47] 胡毅,谭北平,麦康森,等.饲料中益生菌对凡纳滨对虾生长、肠道菌群及部分免疫指标的影响[J].中国水产科学,2008,15(2):244-251. [48] ATARASHI K, SUDA W, LUO C W, et al. Ectopic colonization of oral bacteria in the intestine drives TH1 cell induction and inflammation[J].Science,2017,358(6361):359-365. [49] MAZMANIAN S K, ROUND J L, KASPER D L. A microbial symbiosis factor prevents intestinal inflammatory disease[J].Nature,2008,453(7195):620-625.
2023, Vol. 42 
