凡纳滨对虾急性肝胰腺坏死综合症病虾与健康虾肠道优势菌群比较分析
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摘要
为研究急性肝胰腺坏死综合征(Acute hepatopancreatic necrosis syndrome,AHPNS)病原对凡纳滨对虾肠道菌群的影响,获取更多的AHPNS防治基础资料,采用PCR-RFLP方法比较了江苏地区患AHPNS病虾和健康对虾肠道微生物群落。结果发现,不同生长阶段病虾肠道中均只有弧菌属细菌;对照的健康对虾中,幼虾肠道优势菌为假单胞菌属、食酸菌属和固氮螺菌属细菌;养殖中期对虾肠道优势菌为发光杆菌属、弧菌属和希瓦氏菌属细菌;养殖后期对虾肠道优势菌为弧菌属、红杆菌属、莱茵海默氏菌属细菌。研究结果表明,凡纳滨对虾患AHPNS后肠道微生物群落明显不同于健康对虾,肠道菌群多样性遭到破坏,弧菌为肠道内绝对优势细菌,不同生长阶段健康对虾的肠道优势菌存在差异。本研究探究了AHPNS病原对于凡纳滨对虾肠道菌群的影响,并为认识和防控AHPNS提供试验基础。
In order to study the effect of acute hepatopancreatic necrosis syndrome( AHPNS) pathogeny on the intestinal microflora of L. vannamei and acquire more basic data for prevention and control of AHPNS,showed the difference between the intestinal microflora of AHPNS positive shrimp and healthy shrimp was compared by PCR-RFLP. The results showed that there were only Vibrio bacteria in intestine of diseased shrimp at different growth stages. Using healthy shrimp as contorl,dominant intestinal bacteria of juvenile shrimp were Pseudomonas,Acidovoraxs and Brevundimonas,and dominant intestinal bacteria of breeding mid-term shrimpwere Vibrio, Photobacterium and Shewanella, dominant intestinal bacteria of breeding late shrimp were Vibrio,Rhodobacteraceae and Rheinheimera. The results showed that the intestinal microbial community of L. vannamei infected with AHPNS was significantly different from that of healthy L. vannamei,and the diversity of intestinal flora was destroyed. Vibrio was the dominant bacteria in the intestine,and there were differences in intestinal dominant bacteria of healthy shrimp at different growth stages. This study explores the effect of AHPNS on intestinal flora,and provides experimental basis for recognition and prevention and control AHPNS.
In order to study the effect of acute hepatopancreatic necrosis syndrome( AHPNS) pathogeny on the intestinal microflora of L. vannamei and acquire more basic data for prevention and control of AHPNS,showed the difference between the intestinal microflora of AHPNS positive shrimp and healthy shrimp was compared by PCR-RFLP. The results showed that there were only Vibrio bacteria in intestine of diseased shrimp at different growth stages. Using healthy shrimp as contorl,dominant intestinal bacteria of juvenile shrimp were Pseudomonas,Acidovoraxs and Brevundimonas,and dominant intestinal bacteria of breeding mid-term shrimpwere Vibrio, Photobacterium and Shewanella, dominant intestinal bacteria of breeding late shrimp were Vibrio,Rhodobacteraceae and Rheinheimera. The results showed that the intestinal microbial community of L. vannamei infected with AHPNS was significantly different from that of healthy L. vannamei,and the diversity of intestinal flora was destroyed. Vibrio was the dominant bacteria in the intestine,and there were differences in intestinal dominant bacteria of healthy shrimp at different growth stages. This study explores the effect of AHPNS on intestinal flora,and provides experimental basis for recognition and prevention and control AHPNS.
引文
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[3] SOTO-RODRIGUEZ S A,GOMEZGIL B,LOZANOOLVERA R,et al. Field and experimental evidence of Vibrio parahaemolyticusas the causative agent of acute hepatopancreatic necrosis disease ofcultured shrimp(Litopenaeus vannamei)in Northwestern Mexico.[J]. Appl Environ Microbiol,2015,81(5):1689-1699.
[4] JOSHI J,SRISALA J,TRUONG V H,et al. Variation in Vibrioparahaemolyticus,isolates from a single Thai shrimp farm experi-encing an outbreak of acute hepatopancreatic necrosis disease(AHPND)[J]. Aquaculture,2014(428/429):297-302.
[5] OXLEY A P A,SHIPTON W,OWENS L,et al. Vacterial florafrom the gut of the wild and cultured banana prawn,Penaeus merg-uiensis[J]. J Appl Microbiol,2002,93(2):214-223.
[6]吴金凤,熊金波,王欣,等.肠道菌群对凡纳滨对虾健康的指示作用[J].应用生态学报,2016,27(2):611-621.
[7]杨慧婷.对虾肠道菌群动态平衡的免疫调控研究[D].济南:山东大学,2016.
[8]张盛静,赵小金,宋晓玲,等.饲料添加益生菌对凡纳滨对虾肠道菌群、Toll受体及溶菌酶基因表达及抗感染的影响[J].中国水产科学,2016,23(4):846-854.
[9]吴定心.微生物制剂对南美白对虾养殖体系微生态的影响及其与藻类关系的研究[D].武汉:华中农业大学,2016.
[10] SIRIKHARIN R,TAENGCHAIYAPHUM S,SANGUANRUT P,et al. Characterization and PCR detection of binary,Pir-Like Tox-ins from Vibrio parahaemolyticus isolates that cause acute hepato-pancreatic necrosis disease(AHPND)in shrimp.[J]. PLoSONE,2015,10(5):219-220.
[11]范志宇,魏后军,仇汝龙,等.一株新型家兔肠炎沙门氏菌的分离鉴定及防治[J].江苏农业学报,2016,32(6):1367-1371.
[12] HAN J E,TANG K F J,ARANGUREN L F,et al. Characteriza-tion and pathogenicity of acute hepatopancreatic necrosis diseasenatural mutants,pir AB vp,(-)V. parahaemolyticus,and pir ABvp,(+)V. campbellii,strains[J]. Aquaculture,2017,470:84-90.
[13] DONG X,BI D,WANG H,et al. pirABvp-Bearing Vibrio parah-aemolyticus and Vibrio campbellii pathogens isolated from the sameAHPND-Affected pond possess highly similar pathogenic plasmids[J]. Frontiers in Microbiology,2017,8:1859.
[14]蒋葛,沈辉,万夕和,等.凡纳滨对虾急性肝胰腺坏死综合征研究进展[J].动物医学进展,2018(4):87-91.
[15] LEE C T,CHEN I T,YANG Y T,et al. Correction for Lee et al.The opportunistic marine pathogen Vibrio parahaemolyticus be-comes virulent by acquiring a plasmid that expresses a deadly tox-in.[J]. Proc Nati Acad Sci,2015,112(39):798-803.
[16]李继秋,谭北平,麦康森.白斑综合征病毒与凡纳滨对虾肠道菌群区系之间关系的初步研究[J].上海海洋大学学报,2006,15(1):109-113.
[17]孙振丽,宣引明,张皓,等.南美白对虾养殖环境及其肠道细菌多样性分析[J].中国水产科学,2016,23(3):594-605.
[18]张云鹏,朱立颖,肖朝耿,等.南美白对虾体表与肠道细菌菌群结构的DGGE分析[J].中国食品学报,2016,16(8):218-224.
[19] ZHENG Y F,YU M,LIU J W,et al. Bacterial community associ-ated with healthy and diseased pacific white shrimp(Litopenaeusvannamei)larvae and rearing water across different growth stages[J]. Front Microbiol,2017,8(18):1-11.
[2] LA PEA L D,CABILLON N A,CATEDRAL D D,et al. Acutehepatopancreatic necrosis disease(AHPND)outbreaks in Penaeusvannamei and P. monodon cultured in the Philippines.[J]. DisAquat Org,2015,116(3):251-254.
[3] SOTO-RODRIGUEZ S A,GOMEZGIL B,LOZANOOLVERA R,et al. Field and experimental evidence of Vibrio parahaemolyticusas the causative agent of acute hepatopancreatic necrosis disease ofcultured shrimp(Litopenaeus vannamei)in Northwestern Mexico.[J]. Appl Environ Microbiol,2015,81(5):1689-1699.
[4] JOSHI J,SRISALA J,TRUONG V H,et al. Variation in Vibrioparahaemolyticus,isolates from a single Thai shrimp farm experi-encing an outbreak of acute hepatopancreatic necrosis disease(AHPND)[J]. Aquaculture,2014(428/429):297-302.
[5] OXLEY A P A,SHIPTON W,OWENS L,et al. Vacterial florafrom the gut of the wild and cultured banana prawn,Penaeus merg-uiensis[J]. J Appl Microbiol,2002,93(2):214-223.
[6]吴金凤,熊金波,王欣,等.肠道菌群对凡纳滨对虾健康的指示作用[J].应用生态学报,2016,27(2):611-621.
[7]杨慧婷.对虾肠道菌群动态平衡的免疫调控研究[D].济南:山东大学,2016.
[8]张盛静,赵小金,宋晓玲,等.饲料添加益生菌对凡纳滨对虾肠道菌群、Toll受体及溶菌酶基因表达及抗感染的影响[J].中国水产科学,2016,23(4):846-854.
[9]吴定心.微生物制剂对南美白对虾养殖体系微生态的影响及其与藻类关系的研究[D].武汉:华中农业大学,2016.
[10] SIRIKHARIN R,TAENGCHAIYAPHUM S,SANGUANRUT P,et al. Characterization and PCR detection of binary,Pir-Like Tox-ins from Vibrio parahaemolyticus isolates that cause acute hepato-pancreatic necrosis disease(AHPND)in shrimp.[J]. PLoSONE,2015,10(5):219-220.
[11]范志宇,魏后军,仇汝龙,等.一株新型家兔肠炎沙门氏菌的分离鉴定及防治[J].江苏农业学报,2016,32(6):1367-1371.
[12] HAN J E,TANG K F J,ARANGUREN L F,et al. Characteriza-tion and pathogenicity of acute hepatopancreatic necrosis diseasenatural mutants,pir AB vp,(-)V. parahaemolyticus,and pir ABvp,(+)V. campbellii,strains[J]. Aquaculture,2017,470:84-90.
[13] DONG X,BI D,WANG H,et al. pirABvp-Bearing Vibrio parah-aemolyticus and Vibrio campbellii pathogens isolated from the sameAHPND-Affected pond possess highly similar pathogenic plasmids[J]. Frontiers in Microbiology,2017,8:1859.
[14]蒋葛,沈辉,万夕和,等.凡纳滨对虾急性肝胰腺坏死综合征研究进展[J].动物医学进展,2018(4):87-91.
[15] LEE C T,CHEN I T,YANG Y T,et al. Correction for Lee et al.The opportunistic marine pathogen Vibrio parahaemolyticus be-comes virulent by acquiring a plasmid that expresses a deadly tox-in.[J]. Proc Nati Acad Sci,2015,112(39):798-803.
[16]李继秋,谭北平,麦康森.白斑综合征病毒与凡纳滨对虾肠道菌群区系之间关系的初步研究[J].上海海洋大学学报,2006,15(1):109-113.
[17]孙振丽,宣引明,张皓,等.南美白对虾养殖环境及其肠道细菌多样性分析[J].中国水产科学,2016,23(3):594-605.
[18]张云鹏,朱立颖,肖朝耿,等.南美白对虾体表与肠道细菌菌群结构的DGGE分析[J].中国食品学报,2016,16(8):218-224.
[19] ZHENG Y F,YU M,LIU J W,et al. Bacterial community associ-ated with healthy and diseased pacific white shrimp(Litopenaeusvannamei)larvae and rearing water across different growth stages[J]. Front Microbiol,2017,8(18):1-11.