首页 >  大连海洋大学学报 >  一株虾源假交替单胞菌(Pseudoalteromonas sp.)的分离鉴定及口服后对对虾体内弧菌的影响

2023, 38(2): 223-232. doi: 10.16535/j.cnki.dlhyxb.2022-150

一株虾源假交替单胞菌(Pseudoalteromonas sp.)的分离鉴定及口服后对对虾体内弧菌的影响

1. 浙江海洋大学 水产学院, 浙江 舟山 316022;

2. 中国水产科学研究院黄海水产研究所 农业农村部海水养殖病害防治重点实验室, 青岛市海水养殖流行病学与生物安保重点实验室, 山东 青岛 266071;

3. 海南中正水产科技有限公司, 海南 东方 572632

收稿日期:2022-05-10

基金项目:   国家重点研发计划"蓝色粮仓科技创新"项目(2019YFD0900105)  "科技助力经济2020"重点专项(SQ2020YFF0425682)  中国水产科学研究院黄海水产研究所基本科研业务费(20603022018001) 

关键词: 凡纳滨对虾 , 假交替单胞菌 , 拮抗菌 , 对虾病原菌

Isolation, identification and effects of a strain of Pseudoalteromonas sp. on Vibrio sp. in shrimp by oral administration

1. Fisheries College, Zhejiang Ocean University, Zhoushan 316022, China;

2. Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;

3. Hainan Zhongzheng Aquatic Science and Technology Company Limited, Dongfang 572632, China

Received Date:2022-05-10

Keywords: Litopenaeus vannamei , Pseudoalteromonas sp. , antagonistic bacterium , shrimp pathogenic bacterium

摘要

为探讨对虾细菌性病原的微生物防控技术,采用牛津杯打孔法对分离自对虾育苗池的一株编号为2021041402-14的细菌 (简称"02HN")进行了抗菌测试,分析其对欧文氏弧菌(Vibrio owensii)、哈维氏弧菌(V.harveyi)、坎贝氏弧菌(V.campbellii)、溶藻弧菌(V.alginolyticus)、副溶血弧菌(V.parahaemolyticus)和美人鱼发光杆菌(Photobacterium damselae) 等6株虾病原菌的抑菌效果及最低抑菌浓度,通过浸泡感染测试了菌株02HN对凡纳滨对虾(Litopenaeus vannamei)的生物毒性,采用16S rRNA、gyrB-rpoD串联基因序列比对法及生理生化方法对该菌进行了鉴定,并将该菌添加到饲料中投喂对虾,评价该菌对对虾体内弧菌数量的影响。结果表明:菌株02HN对欧文氏弧菌、哈维氏弧菌、坎贝氏弧菌、溶藻弧菌、副溶血弧菌和美人鱼发光杆菌均具有抑菌效果,其最低抑菌浓度分别为2.32×104、2.32×103、2.32×106、2.32×106、2.32×103、2.32×106 CFU/mL;采用02HN浓度为1×107 CFU/mL以下的菌液浸泡凡纳滨对虾,生物毒性不明显;分子鉴定显示,该菌与杀鱼假交替单胞菌(Pseudoalteromonas piscicida)及金丽假交替单胞菌(P.flavipulchra)最为接近,为一株假交替单胞菌(Pseudoalteromonas sp.);对虾摄食菌含量分别为1×105、1×107cells/g的饵料后,其存活及生长无显著性变化,但能显著降低对虾肝胰腺中的弧菌数量(P<0.05)。研究表明,菌株02HN在对虾养殖中具有开发应用潜能,本研究结果可为对虾细菌性病原的生物防控提供生物材料及技术思路。

为探讨对虾细菌性病原的微生物防控技术,采用牛津杯打孔法对分离自对虾育苗池的一株编号为2021041402-14的细菌 (简称"02HN")进行了抗菌测试,分析其对欧文氏弧菌(Vibrio owensii)、哈维氏弧菌(V.harveyi)、坎贝氏弧菌(V.campbellii)、溶藻弧菌(V.alginolyticus)、副溶血弧菌(V.parahaemolyticus)和美人鱼发光杆菌(Photobacterium damselae) 等6株虾病原菌的抑菌效果及最低抑菌浓度,通过浸泡感染测试了菌株02HN对凡纳滨对虾(Litopenaeus vannamei)的生物毒性,采用16S rRNA、gyrB-rpoD串联基因序列比对法及生理生化方法对该菌进行了鉴定,并将该菌添加到饲料中投喂对虾,评价该菌对对虾体内弧菌数量的影响。结果表明:菌株02HN对欧文氏弧菌、哈维氏弧菌、坎贝氏弧菌、溶藻弧菌、副溶血弧菌和美人鱼发光杆菌均具有抑菌效果,其最低抑菌浓度分别为2.32×104、2.32×103、2.32×106、2.32×106、2.32×103、2.32×106 CFU/mL;采用02HN浓度为1×107 CFU/mL以下的菌液浸泡凡纳滨对虾,生物毒性不明显;分子鉴定显示,该菌与杀鱼假交替单胞菌(Pseudoalteromonas piscicida)及金丽假交替单胞菌(P.flavipulchra)最为接近,为一株假交替单胞菌(Pseudoalteromonas sp.);对虾摄食菌含量分别为1×105、1×107cells/g的饵料后,其存活及生长无显著性变化,但能显著降低对虾肝胰腺中的弧菌数量(P<0.05)。研究表明,菌株02HN在对虾养殖中具有开发应用潜能,本研究结果可为对虾细菌性病原的生物防控提供生物材料及技术思路。

参考文献

[1] 农业农村部渔业渔政管理局,全国水产技术推广总站,中国水产学会编制.2021中国渔业统计年鉴[M].北京:中国农业出版社,2021. Fishery Administration Bureau of Ministry of Agriculture and Rural Affairs,National Fisheries Technology Extension Station,China Society of Fisheries.2021 China fisheries statistical yearbook[M].Beijing:China Agriculture Press,2021.(in Chinese)
[2] DE LA PEÑA L D,CABILLON N A R,CATEDRAL D D,et al.Acute hepatopancreatic necrosis disease (AHPND) outbreaks in Penaeus vannamei and P. monodon cultured in the Philippines[J].Diseases of Aquatic Organisms,2015,116(3):251-254.
[3] ZOU Y,XIE G S,JIA T C,et al.Determination of the infectious agent of translucent post-larva disease (TPD) in Penaeus vannamei[J].Pathogens,2020,9(9):741.
[4] ZHANG D H,BADER T J,LANGE M D,et al.Toxicity of recombinant PirA and PirB derived from Vibrio parahaemolyticus in shrimp[J].Microbial Pathogenesis,2021,155:104886.
[5] ROBERTSON P,CALDERON J,CARRERA L,et al.Experimental Vibrio harveyi infections in Penaeus vannamei larvae[J].Diseases of Aquatic Organisms,1998,32:151-155.
[6] THIRUGNANASAMBANDAM R,INBAKANDAN D,ABRAHAM L S, et al.De novo assembly and annotation of the whole genomic analysis of Vibrio campbellii RT-1 strain,from infected shrimp:Litopenaeus vannamei[J].Microbial Pathogenesis,2017,113:372-377.
[7] LIU L Y,XIAO J Z,ZHANG M M,et al.A Vibrio owensii strain as the causative agent of AHPND in cultured shrimp,Litopenaeus vannamei[J].Journal of Invertebrate Pathology,2018,153:156-164.
[8] NGO H V,HUANG H T,LEE P T,et al.Effects of Phyllanthus amarus extract on nonspecific immune responses,growth,and resistance to Vibrio alginolyticus in white shrimp Litopenaeus vannamei[J].Fish & Shellfish Immunology,2020,107(Pt A):1-8.
[9] AKBARY P,ADESHINA I,JAHANBAKHSHI A.Growth performance,digestive enzymes,antioxidant activity and immune responses of Litopenaeus vannamei fed with Jania adhaerens J.V.supplemented diet against Photobacterium damselae infection[J].Animal Feed Science and Technology,2020,270:114696.
[10] PANTOJAC R,LIGHTNER D V,NOBLE B L,et al.Early mortality syndrome affects shrimp in Asia[J].Global Aquaculture Advocate,2012,15(1):40.
[11] THITAMADEE S,PRACHUMWAT A,SRISALA J,et al.Review of current disease threats for cultivated penaeid shrimp in Asia[J].Aquaculture,2016,452:69-87.
[12] LEYVA-MADRIGAL K Y,LUNA-GONZÁLEZ A,ESCOBEDO-BONILLA C M,et al.Screening for potential probiotic bacteria to reduce prevalence of WSSV and IHHNV in whiteleg shrimp (Litopenaeus vannamei) under experimental conditions[J].Aquaculture,2011,322/323:16-22.
[13] 张欢欢,王秀华,李晨,等.对虾养殖池中一株弧菌拮抗菌的分离鉴定[J].渔业科学进展,2016,37(3):85-92. ZHANG H H,WANG X H,LI C,et al.Isolation and identification of a bacterial strain with vibrios-antagonism from shrimp ponds[J].Progress in Fishery Sciences,2016,37(3):85-92.(in Chinese)
[14] 于敏.金丽假交替单胞菌JG1抑菌机理研究[D].青岛:中国海洋大学,2013. YU M.The antibacterial mechanism of Pseudoalteromonas flavipulchra JG1[D].Qingdao:Ocean University of China,2013.(in Chinese)
[15] 张宇哲,王秀华,于党辉,等.抗弧菌杀鱼假交替单胞菌2515的潜在毒性及热处理脱毒后抗菌效果[J].大连海洋大学学报,2021,36(3):399-405. ZHANG Y Z,WANG X H,YU D H,et al.Potential toxicity,antibacterial effect and antagonistic activity against Vibrio of a strain of bacterium Pseudoalteromonas piscicida 2515 exposed to heat treatment detoxification[J].Journal of Dalian Ocean University,2021,36(3):399-405.(in Chinese)
[16] 李静舒,张恒慧,贺东亮.高活菌数微生态制剂的制备及其抑菌活性研究[J].饲料研究,2020,43(1):96-100. LI J S,ZHANG H H,HE D L.Study on preparation and antibacterial activity of probiotics with high viable count[J].Feed Research,2020,43(1):96-100.(in Chinese)
[17] 李成海,贠建民,艾对元,等.金黄色葡萄球菌拮抗菌株的筛选及鉴定[J].食品工业科技,2013,34(7):195-198. LI C H,YUN J M,AI D Y,et al.Screening and identification of bacterial strain against Staphyloccocus aureus[J].Science and Technology of Food Industry,2013,34(7):195-198.(in Chinese)
[18] YÁÑEZ M A,CATALÁN V,APRÁIZ D,et al.Phylogenetic analysis of members of the genus Aeromonas based on gyrB gene sequences[J].International Journal of Systematic and Evolutionary Microbiology,2003,53(Pt 3):875-883.
[19] SENDEROVICH Y,KEN-DROR S,VAINBLAT I,et al.A molecular study on the prevalence and virulence potential of Aeromonas spp.recovered from patients suffering from diarrhea in Israel[J].PLoS One,2012,7(2):e30070.
[20] KHALIFA A,ALDAYEL M.Isolation and characterisation of the agarolytic bacterium Pseudoalteromonas ruthenica[J].Open Life Sciences,2019,14:588-594.
[21] 王枫林,王秀华,张宇哲,等.杀鱼假交替单胞菌2515的抗弧菌效果及在对虾养殖中的应用[J].中国水产科学,2021,28(7):903-913. WANG F L,WANG X H,ZHANG Y Z,et al.Anti-Vibrio effect of Pseudoalteromonas piscicida 2515 and its application in shrimp culture[J].Journal of Fishery Sciences of China,2021,28(7):903-913.(in Chinese)
[22] 王磊.脱氮菌的筛选及在凡纳滨对虾生物絮团养殖中的应用[D].上海:上海海洋大学,2015. WANG L.The selection and application of denitrifying bacteria in Litopenaeus vannamei biological floccules breeding[D].Shanghai:Shanghai Ocean University,2015.(in Chinese)
[23] GAUTHIER G,GAUTHIER M,CHRISTEN R.Phylogenetic analysis of the genera Alteromonas,Shewanella, and Moritella using genes coding for small-subunit rRNA sequences and division of the genus Alteromonas into two genera,Alteromonas (emended) and Pseudoalteromonas gen.nov.,and proposal of twelve new species combinations[J].International Journal of Systematic Bacteriology,1995,45(4):755-761.
[24] BOWMAN J P. Bioactive compound synthetic capacity and ecological significance of marine bacterial genus Pseudoalteromonas[J].Marine Drugs,2007,5(4):220-241.
[25] ZHOU M Y,CHEN X L,ZHAO H L,et al.Diversity of both the cultivable protease-producing bacteria and their extracellular proteases in the sediments of the South China Sea[J].Microbial Ecology,2009,58(3):582-590.
[26] HUANG Y L,LI M,YU Z L,et al.Correlation between pigmentation and larval settlement deterrence by Pseudoalteromonas sp.sf57[J].Biofouling,2011,27(3):287-293.
[27] YU M,TANG K H,LIU J W,et al.Genome analysis of Pseudoalteromonas flavipulchra JG1 reveals various survival advantages in marine environment[J].BMC Genomics,2013,14:707.
[28] WANG H L,WANG C D,TANG Y,et al.Pseudoalteromonas probiotics as potential biocontrol agents improve the survival of Penaeus vannamei challenged with acute hepatopancreatic necrosis disease (AHPND)-causing Vibrio parahaemolyticus[J].Aquaculture,2018,494:30-36.
[29] RICHARDS G P,WATSON M A,NEEDLEMAN D S,et al.Mechanisms for Pseudoalteromonas piscicida-induced killing of vibrios and other bacterial pathogens[J].Applied and Environmental Microbiology,2017,83(11):e00175-e00117.
[30] 席宇,朱大恒,刘红涛,等.假交替单胞菌及其胞外生物活性物质研究进展[J].微生物学通报,2005,32(3):108-112. XI Y,ZHU D H,LIU H T,et al.Advances on Pseudoalteromonas species and their extracellular bioactive compounds[J].Microbiology,2005,32(3):108-112.(in Chinese)
[31] 王印庚,方波,张春云,等.养殖刺参保苗期重大疾病"腐皮综合征"病原及其感染源分析[J].中国水产科学,2006,13(4):610-616. WANG Y G,FANG B,ZHANG C Y,et al.Etiology of skin ulcer syndrome in cultured juveniles of Apostichopus japonicus and analysis of reservoir of the pathogens[J].Journal of Fishery Sciences of China,2006,13(4):610-616.(in Chinese)
[32] 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. HU Y,TAN B P,MAI K S,et al.Effects of dietary probiotic on growth,immunity and intestinal bacteria of juvenile Litopenaeus vannamei[J].Journal of Fishery Sciences of China,2008,15(2):244-251.(in Chinese)
[33] CRAB R,DEFOIRDT T,BOSSIER P,et al.Biofloc technology in aquaculture:beneficial effects and future challenges[J].Aquaculture,2012,356/357:351-356.

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目录

一株虾源假交替单胞菌(Pseudoalteromonas sp.)的分离鉴定及口服后对对虾体内弧菌的影响