高发性拟穴青蟹细菌病的病原分离及鉴定
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摘要
从患病拟穴青蟹(Scylla paramamosain)肝胰腺中分离到2种优势菌株,对2种优势菌株进行了鉴定和特性分析。利用形态学观察、生理生化特性分析和16S r DNA序列测定法对细菌类别进行判定,并采用药敏试验对细菌的耐药性进行分析。结果表明,这2种细菌分别为弧菌(Vibrio sp.)和希瓦氏菌(Shewanella),利用15种抗生素对2种细菌进行药敏试验分析,发现2种细菌对恩诺沙星、氟苯尼考、强力霉素高度敏感,对其他几种抗生素则有一定的耐受性。研究进一步证实了海水致病性弧菌是拟穴青蟹细菌性疾病高发的主要原因,也表明希瓦氏菌可能是一种潜在病原菌,与弧菌协同作用,对拟穴青蟹规模化养殖造成危害,同时也为拟穴青蟹养殖过程中细菌性病害的确定和防治提供参考依据。
Two dominant bacteria were isolated from the hepatopancreas of the diseased mud crab(Scylla paramamosain).The isolated strains were identified as Vibrio sp. and Shewanella through analyzing phenotypic characteristics and 16 S rDNA sequence. Antibiotic sensitivity patterns of the two kinds of bacteria were evaluated against 15 antibiotics. Results showed the the isolated strain was highly sensitivity to 3 antibiotics including enrofloxacin, florfenicol and doxycycline, but resistant to 12 other antibiotics.
Two dominant bacteria were isolated from the hepatopancreas of the diseased mud crab(Scylla paramamosain).The isolated strains were identified as Vibrio sp. and Shewanella through analyzing phenotypic characteristics and 16 S rDNA sequence. Antibiotic sensitivity patterns of the two kinds of bacteria were evaluated against 15 antibiotics. Results showed the the isolated strain was highly sensitivity to 3 antibiotics including enrofloxacin, florfenicol and doxycycline, but resistant to 12 other antibiotics.
引文
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[19]张晓华,廖绍安,李筠,等.海湾扇贝病原菌(漂浮弧菌)的研究[J].青岛海洋大学学报,1998,28(3):426-432.
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[21] SAEED M O,ALAMOUDI M M,AL-HARBI A H. A Pseudomonas associated with disease in cultured rabbitfish Siganus rivulatus in the Red Sea[J].Dis Aquat Organ,1987,3(3):177-180.
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[23]贾春红,李淑芳,张继东.3株方斑东风螺病原希瓦氏菌对35种常用抗菌药物的药敏性试验[J].江西水产科技,2013(1):17-19.
[24]孙琪,邱军强,杨先乐.希瓦氏菌引起的海马细菌性疾病[A].中国水产学会.2012年中国水产学会学术年会论文摘要集[C].北京:2012.
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[26] PEKALA A,KOZINSKA A,PAZDZIOR E,et al. Phenotypical and genotypical characterization of Shewanella putrefaciens strains isolated from diseased freshwater fish[J].J Fish Dis,2015,38(3):283-293.
[27]秦蕾,张晓君,毕可然.一种新的异育银鲫病原—腐败希瓦氏菌[J].微生物学报,2012,52(5):558-565.
[28]沈萍,范秀容,李广武.微生物实验[M].第三版.北京:高等教育出版社,1999.62-64.
[29] M100-S18-2008,National Committee for Clinical Laboratory Standards. Performance standards for antimicrobial susceptibility testing[S].
[30] KIM M S,JEONG H D. Development of 16S rRNA targeted PCR methods for the detection and differentiation of Vibrio vulnificus in marine environments[J].Aquaculture,2001,193(3-4):199-211.
[31] PAUNGFOO C,PRASERTSAN P,BURRELL P C,et al. Nitrifying bacterial communities in an aquaculture wastewater treatment system using fluorescence in situ hybridization(FISH),16S rRNA gene cloning,and phylogeneticanalysis[J].Biotechn Bioeng,2007,97(4):985-990.
[2] JITHENDRAN K P,POORNIMA M,BALASUBRANMANIAN C P,et al. Diseases of mud crabs(Scylla spp.):An overview[J].Indian J Fish,2010,57(3):55-63.
[3]郑天伦,黄家庆,孔蕾,等.浙江省锯缘青蟹养殖病害调查[J].海洋学研究,2009,27(1):96-100.
[4]郑天伦.拟穴青蟹病害研究进展[J].水产科学,2014,33(5):326-331.
[5]纪荣兴,黄少涛.锯缘青蟹“黄体病”病原菌的研究[J].台湾海峡,1998,17(4):373-376.
[6]毛芝娟,卓华龙.锯缘青蟹细菌性传染病的病原菌研究[J].水产科学,2001,20(1):8-11.
[7]夏小安,吴清洋,李远友,等.锯缘青蟹混合感染症致病菌的分离鉴定与感染治疗[J].热带海洋学报,2010,29(5):103-110.
[8]王海潮,张兴桃,廖红艳,等.锯缘青蟹黄水病病原菌的分离与鉴定[J].安徽农业科学,2011,39(33):20523-20524,20651.
[9]殷战,徐伯亥.鱼类细菌性疾病的研究[J].水生生物学报,1995,19(1):76-83.
[10]胡超群,陶保华.对虾弧菌病及其免疫预防的研究进展[J].热带海洋,2000,19(3):84-94.
[11]吴后波,潘金培.弧菌属细菌及其所致海水养殖动物疾病[J].中国水产科学,2001,8(1):89-93.
[12]常建波,宫向红,孙逢贤,等.养殖牙鲆弧菌病病原菌初步研究[J].海洋水产研究,2001,22(1):37-41.
[13]王国良,金珊,薛良义,等.海水网箱养殖鲈鱼皮肤溃疡病及其病原菌的研究[J].黄渤海海洋,2000,18(3):85-89.
[14]林克冰,周宸,刘家富.海水网箱养殖大黄鱼病原菌研究[J].海洋科学,1999(4):58-62.
[15]郑国兴,李何,黄宁宁,等.文蛤病原菌(溶藻弧菌)的分离与形状及病文蛤组织的电镜观察[J].水产学报,1991,15(2):85-95.
[16]沈亚林,于业绍.副溶血弧菌对文蛤的致病性及其防治[J].水产学报,1993,17(3):249-252.
[17] BARTLY C H,SLANELZ L W. Occurrence of Vibrio parahaemo lyticus in estuarine waters and oysters of New Hampshire[J].Appl Microbiol,1971,21:965-966.
[18] LACOSTE A,JALABERT F,MALHAM S K,et al. Stress and stress-induced neuroendocrine changes increase the susceptibility of juvenile oysters(Crassostrea gigas)to Vibrio splendidus[J].Appl Envion Microbiol,2001,67(5):2304-2309.
[19]张晓华,廖绍安,李筠,等.海湾扇贝病原菌(漂浮弧菌)的研究[J].青岛海洋大学学报,1998,28(3):426-432.
[20] KORUN J,AKGUN-DAR K,YAZICI M. Isolation of Shewanella putrefaciens from cultured European sea bass,(Dicetrarcbus labrax)in Turkey[J].Revue de Médecine Vétérinaire,2009,160(11):532-536.
[21] SAEED M O,ALAMOUDI M M,AL-HARBI A H. A Pseudomonas associated with disease in cultured rabbitfish Siganus rivulatus in the Red Sea[J].Dis Aquat Organ,1987,3(3):177-180.
[22]吕俊超.养殖大菱鲆病原菌的分离鉴定,组织病理学及免疫组织化学[D].山东青岛:中国海洋大学,2008.
[23]贾春红,李淑芳,张继东.3株方斑东风螺病原希瓦氏菌对35种常用抗菌药物的药敏性试验[J].江西水产科技,2013(1):17-19.
[24]孙琪,邱军强,杨先乐.希瓦氏菌引起的海马细菌性疾病[A].中国水产学会.2012年中国水产学会学术年会论文摘要集[C].北京:2012.
[25] KOZINSKA A,PEKALA A. First isolation of Shewanella putrefaciens from freshwater fish-a potential new pathogen of fish[J].Bulletin-European Association of Fish Pathologists,2004,24(4):189-193.
[26] PEKALA A,KOZINSKA A,PAZDZIOR E,et al. Phenotypical and genotypical characterization of Shewanella putrefaciens strains isolated from diseased freshwater fish[J].J Fish Dis,2015,38(3):283-293.
[27]秦蕾,张晓君,毕可然.一种新的异育银鲫病原—腐败希瓦氏菌[J].微生物学报,2012,52(5):558-565.
[28]沈萍,范秀容,李广武.微生物实验[M].第三版.北京:高等教育出版社,1999.62-64.
[29] M100-S18-2008,National Committee for Clinical Laboratory Standards. Performance standards for antimicrobial susceptibility testing[S].
[30] KIM M S,JEONG H D. Development of 16S rRNA targeted PCR methods for the detection and differentiation of Vibrio vulnificus in marine environments[J].Aquaculture,2001,193(3-4):199-211.
[31] PAUNGFOO C,PRASERTSAN P,BURRELL P C,et al. Nitrifying bacterial communities in an aquaculture wastewater treatment system using fluorescence in situ hybridization(FISH),16S rRNA gene cloning,and phylogeneticanalysis[J].Biotechn Bioeng,2007,97(4):985-990.