杂交鲟源恶臭假单胞菌的分离鉴定及药敏特性研究
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摘要
对患肠炎病杂交鲟进行病原分离、鉴定及药敏试验,为杂交鲟肠炎病的防控提供参考。从患病杂交鲟肝、肾、脾及肠道中分离纯化病原菌,经理化特性测定及16S rRNA序列分析对其进行鉴定,开展人工感染试验,并利用K-B法进行药敏特性分析。结果显示,分离到的XY4菌株是本次引发杂交鲟病害的致病菌,其对杂交鲟的LD_(50)为1.30×10~6cfu·g~(-1)。XY4菌株理化特性与恶臭假单胞菌一致,16S rRNA序列与恶臭假单胞菌同源性为100%,综合判定XY4菌株为恶臭假单胞菌。XY4菌株对头孢拉定、氟苯尼考及多西环素等9种抗生素高度敏感。养殖时可选用对恶臭假单胞菌敏感药物进行防控。
The isolation,identification and antibiotic sensitivity of pathogen from the morbid hybrid sturgeon were conducted to provide a reference for control and prevention disease for hybrid sturgeon. The pathogenic bacteria were isolated and purified from liver,kidney,spleen and intestinal tract of hybrid sturgeon. The biochemical identification and 16S rRNA gene sequence determination and the artificial infection test were used to identify whether the strain XY4 was the pathogen,and the antimicrobial susceptibility test was conducted by K-B method. The results showed that strain XY4 was the pathogens of hybrid sturgeon,the LD_(50) of the isolate was counted to 1. 30 × 10~6 cfu·g~(-1). According to morphological and biochemical characteristics as well as the result of 16S rRNA gene sequence analysis,the isolated strain XY4 was Pseudomonas putida. Strain XY4 was susceptible to cefadroxil,florfenicol,doxycycline and other 6 kinds of antibiotics. The disease caused by Pseudomonas putida can be prevented by above sensitive drugs in fishery farming.
The isolation,identification and antibiotic sensitivity of pathogen from the morbid hybrid sturgeon were conducted to provide a reference for control and prevention disease for hybrid sturgeon. The pathogenic bacteria were isolated and purified from liver,kidney,spleen and intestinal tract of hybrid sturgeon. The biochemical identification and 16S rRNA gene sequence determination and the artificial infection test were used to identify whether the strain XY4 was the pathogen,and the antimicrobial susceptibility test was conducted by K-B method. The results showed that strain XY4 was the pathogens of hybrid sturgeon,the LD_(50) of the isolate was counted to 1. 30 × 10~6 cfu·g~(-1). According to morphological and biochemical characteristics as well as the result of 16S rRNA gene sequence analysis,the isolated strain XY4 was Pseudomonas putida. Strain XY4 was susceptible to cefadroxil,florfenicol,doxycycline and other 6 kinds of antibiotics. The disease caused by Pseudomonas putida can be prevented by above sensitive drugs in fishery farming.
引文
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[2]崔禾,何建湘,郑维中.我国鲟鱼产业现状分析及发展建议[J].中国水产,2006(6):8-10.CUI H,HE J X,ZHENG W Z.Analysis and development proposals about the status of the our sturgeon industry[J].China Fisheries,2006(6):8-10.(in Chinese)
[3]田甜,杨元金,王京树,等.鲟鱼病害研究进展[J].湖北农业科学,2012,51(3):559-563TIAN T,YANG Y J,WANG J S,et al.Research progress in sturgeon diseases[J].Hubei Agricultural Sciences,2012,51(3):559-563.(in Chinese)
[4]李圆圆,曹海鹏,何姗,等.鲟源致病性嗜水气单胞菌X1的分离鉴定与药敏特性研究[J].微生物学通报.2008,35(8):1186-1191.LI Y Y,CAO H P,HE S,et al.Isolation and identification of Aeromonas hydrophila strain X1 from Acipenser baerii and its antibiotic sensitivity[J].Microbiology China,2008,35(8):1186-1191.(in Chinese with English abstract)
[5]赵凤岐,曹谨玲,刘青.西伯利亚鲟败血症病理学观察与病原学研究[J].水生生物学报,2009,33(2):316-323.ZHAO F Q,CAO J L,LIU Q.Study on pathology and etiology of hemorrhagic septicemia in Acipenser baerii[J].Acta Hydrobiology Sinica,2009,33(2):316-323.(in Chinese with English abstract)
[6]曹海鹏,杨先乐,高鹏,等.鲟细菌性败血综合征致病菌的初步研究[J].淡水渔业,2007,37(2):53-56.CAO H P,YANG X L,GAO P,et al.Preliminary study of the pathogens isolated from bacterial septicaemia syndrome of sturgeon[J].Freshwater Fisheries,2007,37(2):53-56.(in Chinese with English abstract)
[7]王小亮,徐立蒲,曹欢,等.鲟致病性类志贺邻单胞菌的鉴定、致病性及药敏特性[J].微生物学报,2013,53(7):723-729.WANG X L,XU L P,CAO H,et al.Identification and drug sensitivity of a Plesiomonas shigelloides from diseased sturgeons[J].Acta Microbiologica Sinica,2013,53(7):723-729.(in Chinese with English abstract)
[8]潘厚军,刘晓勇,常藕琴,等.西伯利亚鲟停乳链球菌的分离、鉴定与致病性[J].中国水产科学,2009,16(6):891-904.PAN H J,LIU X Y,CHANG O Q,et al.Isolation,identification and pathogenicity of Streptococcus dysgalactiae from Siberians turgeon,Acipenser baerii[J].Journal of Fishery Sciences of China,2009,16(6):891-904.(in Chinese with English abstract)
[9]王小亮,徐立蒲,王静波,等.杂交鲟海豚链球菌的分离、鉴定及药物敏感性[J].微生物学报,2014,54(4):442-448.WANG X L,XU L P,WANG J B,et al.Isolation,identification and drug sensitivity of Streptococcus iniae from hybrid sturgeons(Huso dauricus female×Acipenser schrencki male)[J].Acta Microbiologica Sinica,2014,54(4):442-448.(in Chinese with English abstract)
[10]XU J,ZENG X,JIANG N,et al.Pseudomonas alcaligenes infection and mortality in cultured Chinese sturgeon,Acipenser sinensis[J].Aquaculture,2015,446:37-41.
[11]肖艳翼,王斌,夏永涛,等.鲟病原性温和气单胞菌的分离鉴定及药敏试验[J].南方农业学报,2015,46(10):1909-1914.XIAO Y Y,WANG B,XIA Y T,et al.Isolation,identification and drug sensitivity test of pathogenic Aeromonas sobria from sturgeon[J].Journal of Southern Agriculture,2015,46(10):1909-1914.(in Chinese with English abstract)
[12]杨移斌,夏永涛,郑卫东,等.鲟源鲁氏耶尔森菌的分离鉴定及药敏特性研究[J].水生生物学报,2013,37(2):393-398.YANG Y B,XIA Y T,ZHENG W D,et al.Isolation and identification of Yersinia ruckeri from Acipenser baerii and its antibiotic sensitivity[J].Acta Hydrobiology Sinica,2013,37(2):393-398.(in Chinese with English abstract)
[13]杨移斌,夏永涛,赵蕾,等.鲟源弗氏柠檬酸杆菌分离鉴定及药敏特性研究[J].水生生物学报,2013,37(4):766-771.YANG Y B,XIA Y T,ZHAO L,et al.Isolation and identification of Citrobacter freundii from Amursturgeon(Acipenser schrenckii Brandt)and its antibiotic sensitivity[J].Acta Hydrobiologica Sinica,2013,37(4):766-771.(in Chinese with English abstract)
[14]毛芝娟,毛甬州,汪建萍.两株鱼源琼氏不动杆菌的分离,鉴定和耐药特性分析[J].水产学报,2013,37(10):1572-1578.MAO Z J,MAO Y Z,WANG J P.Isolation,identification and drug-resistance genes detection of Acinetobacter junii from fish[J].Journal of Fisheries of China,2013,37(10):1572-1578.(in Chinese with English abstract)
[15]张德锋,李爱华,龚小宁.鲟分枝杆菌病及其病原研究[J].水生生物学报,2014,38(3):495-504.ZHANG D F,LI A H,GONG X N.Research on the Mycobacterium and its pathogen in sturgeon[J].Acta Hydrobiologica Sinica,2014,38(3):495-504.
[16]徐叔云,卞如濂,陈修.药理实验方法学[M].北京:人民卫生出版社,2002.
[17]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社.2001.
[18]闫茂仓,单乐州,马爱敏,等.条石鲷烂尾白浊症病原菌的分离鉴定及系统发育分析[J].上海海洋大学学报,2010,19(1):50-55.YAN M C,SHAN L Z,MA A M,et al.Identification and phylogenetic analysis of a pathogenic bacterial strain isolated from Oplegnathus fasciatus with tail rot and whitish symptom[J].Journal of Shanghai Ocean University,2010,19(1):50-55.
[19]WIKLER M A.Performance standards for antimicrobial susceptibility testing[M]19th ed.Washington,D.C.,USA:Clinical Laboratory Standards Institute,2009.
[20]杨海洋,李轶,胡洪营,等.恶臭假单胞菌对硝基苯污染河水的修复研究[J].安全与环境工程,2008,15(1):54-57.YANG H Y,LI Y,HU H Y,et al.Bioremediation of nitrobenzene-contaminated river water using Pseudomonas putida[J].Safety and Environmental Engineering,2008,15(1):54-57.(in Chinese with English abstract)
[21]许斌福,程海华,池洪树,等.大黄鱼内脏白点病的病原分析与鉴定[J].福建农业学报,2015(7):631-635.XU B F,CHENG H H,CHI H S,et al.Identification of pathogen responsible for white-spots disease in Larimichthys crocea[J].Fujian Journal of Agricultural Sciences,2015(7):631-635.(in Chinese with English abstract)
[22]樊海平.恶臭假单胞菌引起的欧洲鳗鲡烂鳃病[J].水产学报,2001,25(2):147-150.FAN H P.Study on the gill-rot disease of Anguilla anguilla infected by Pseudomonas putida[J].Journal of Fisheries of China,2001,25(2):147-150.
[23]余为一,李槿年,祖国掌.一株中华绒螯蟹病原菌的研究初报[J].安徽农业大学学报,1999,26(2):174-177.YU W Y,LI J N,ZU G Z.A preliminary report on a pathogenic bacterium from Eriocheir sinensi[J].Journal of Anhui Agricultural University,1999,26(2):174-177.(in Chinese with English abstract)
[24]王高学,黄增荣,袁明.三疣梭子蟹牛奶病病原的分离鉴定[J].西北农林科技大学学报(自然科学版),2007,35(6):29-33WANG G X,HUANG Z R,YUAN M.Isolation and identification of the pathogen of milk disease cultured in swimming crab Portunus tritubercularus(portunidae)[J].Journal of Northwest A&F University(Natural Science Edition),2007,35(6):29-33.(in Chinese with English abstract)
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