泥鳅致病性维氏气单胞菌的分离与鉴定
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摘要
从患病的泥鳅(Misgurnus anguillicaudatus)肝脏中分离得到优势菌NQ150728。对该菌进行了人工感染试验,并利用VITEK-2全自动微生物鉴定仪及16S rRNA、gyr B序列分析对纯培养细菌进行鉴定。此外,对分离的菌株进行药敏实验。结果显示,人工回感试验泥鳅患病症状同自然发病症状,证明其对泥鳅有致病性。VITEK-2全自动微生物鉴定仪鉴定菌株NQ150728为维氏气单胞菌(Aeromonas veronii)。16S rRNA、gyr B基因序列测定结果及系统发育树的构建也表明其为维氏气单胞菌。药敏试验结果表明,该菌对头孢曲松、利福平、恩诺沙星、四环素、氯霉素、头孢哌酮、氧氟沙星、洛美沙星、克拉霉素、氟罗沙星、头孢噻吩、多西环素、新霉素、头孢他啶、大观霉素、环丙沙星、呋喃妥因、头孢拉定、米诺环素、头孢氨苄、氨曲南、左氟沙星、甲氧嘧啶、甲氧胺嘧啶等24种药物高度敏感。
The present study was conducted to clarify species and drug resistance of pathogen from the diseased Misgurnus anguillicaudatus in a farm. Pathogenic bacteria from the liver of the diseased M. anguillicaudatus were examined using conventional methods and isolated. The pathogenic bacteria named strain NQ150728 were isolated and identified as Aeromonas veronii by means of VITEK-2 identification system and 16 S rRNA and gyr B gene sequence analysis. The results of artificial infection test showed that the isolate could induce symptom of natural disease and caused death in 5 days. In addition,strain NQ150728 was highly sensitive to 24 kinds of drug such as ceftriaxone,riftriaxone,enrofloxacin and tetracycline.
The present study was conducted to clarify species and drug resistance of pathogen from the diseased Misgurnus anguillicaudatus in a farm. Pathogenic bacteria from the liver of the diseased M. anguillicaudatus were examined using conventional methods and isolated. The pathogenic bacteria named strain NQ150728 were isolated and identified as Aeromonas veronii by means of VITEK-2 identification system and 16 S rRNA and gyr B gene sequence analysis. The results of artificial infection test showed that the isolate could induce symptom of natural disease and caused death in 5 days. In addition,strain NQ150728 was highly sensitive to 24 kinds of drug such as ceftriaxone,riftriaxone,enrofloxacin and tetracycline.
引文
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[16]ZHU M,WANG X R,LI J,et al.Identification and virulence properties of Aeromonas veronii bv.sobria isolates causing an ulcerative syndrome of loach Misgurnus anguillicaudatus[J].Journal of Fish Diseases,2016,39(6):777-781.
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[18]周勇,曾令兵,李瑞伟,等.患病黄额闭壳龟中产酸克雷伯菌的分离与鉴定[J].淡水渔业,2012,42(3):38-43.ZHOU Y,ZENG L B,LI R W,et al.Isolation and identification of Klebsiella oxytoca from diseased Cuora galbinifrons[J].Freshwater Fisheries,2012,42(3):38-43.(in Chinese with English abstract)
[19]安伟,肖雨,高晓华,等.鳜源致病性荧光假单胞菌的分离与鉴定[J].动物学杂志,2014,49(5):760-765.AN W,XIAO Y,GAO X H,et al.Isolation and identification of pathogenic Pseudomonas fluorescens from Siniperca chuatsi[J].Chinese Journal of Zoology,2014,49(5):760-765.(in Chinese with English abstract)
[20]吴会桃,蔡芷荷,吴清平,等.细菌鉴定系统的应用研究进展[J].中国卫生检验杂志,2010,20(9):2381-2384.WU H T,CAI Z H,WU Q P,et al.Advances in the application of bacterial identification system[J].Chinese Journal of Health Laboratory Technology,2010,20(9):2381-2384.(in Chinese)
[2]肖克宇,蒋武成,李年文,等.泥鳅腐皮病的病原与防治的研究[J].湖南农学院学报,1992,18(增刊l):20-25.XIAO K Y,JIANG W C,LI N W,et al.Stigmatosis of Misgurnus anguillicaudatus and its pathogenic bacteria[J].Journal of Hunan Agricultural college,1992,18(Sl):20-25.(in Chinese with English abstract)
[3]杨鸢劼,陈辉,金日奉,等.泥鳅溃疡病的病原菌分离和组织病理学观察[J].集美大学学报(自然科学版),2004,9(1):22-25.YANG Y J,CHEN H,JIN R F,et al.Isolation of the pathogens and histopathological study on ulcer disease of loach[J].Journal of Jimei University(Natural Science),2004,9(1):22-25.(in Chinese with English abstract)
[4]房海,陈翠珍,张晓君,等.泥鳅气单胞菌感染的检验与分析[J].中国人兽共患病杂志,2006,22(11):1065-1069.FANG H,CHEN C Z,ZHANG X J,et al.Examination and analysis on an infection due to Aeromonas misgurnus[J].Chinese Journal of Zoonoses,2006,22(11):1065-1069.(in Chinese with English abstract)
[5]秦蕾,徐静,张晓君,等.泥鳅的凡隆气单胞菌感染[J].中国人兽共患病杂志,2008,24(12):1100-1102.QIN L,XU J,ZHANG X J,et al.Infection with Aeromonas veronii biovar.in Misgurnus anguillicaudatus[J].Chinese Journal of Zoonoses,2008,24(12):1100-1102.(in Chinese with English abstract)
[6]POLZ M F,CAVANAUGH C M.Bias in template to product ratios in multitemplate PCR[J].Applied&Environmental Microbiology,1998,64(10):3724-3730.
[7]YEZ M A,CATALN V,APRIZ D,et al.Phylogenetic analysis of members of the genus Aeromonas based on gyr B gene sequences[J].International Journal of Systematic and Evolutionary Microbiology,2003,53:875-883.
[8]HOLT J G,KRIEG N R,SNEATH P H A,et al.Bergey's manual of determinative bacteriology[M].9th Ed.London:Williams and Wilkins,Baltimore,1994:190-191,253.
[9]KRIEG N R,HOLT J G.Bergey's manual of systematic bacteriology[M].London:Williams and Wilkins,Baltimore,1984:545-548.
[10]叶应妩,王毓三.全国临床检验操作规程[M].2版.南京:东南大学出版社,1997:553-562.
[11]徐洋,蔺凌云,姚嘉赟,等.黄颡鱼“溃疡综合征”病原的分离鉴定及药敏试验[J].淡水渔业,2015,45(5):100-104.XU Y,LING L Y,YAO J Y,et al.Pathogen isolation,identification and susceptibility test of ulcerative disease syndrome on Pelteobagrus fulvidraco[J].Freshwater Fisheries,2015,45(5):100-104.(in Chinese with English abstract)
[12]房海,陈翠珍,张晓君,等.中华绒螯蟹病原维氏气单胞菌的检验[J].中国人兽共患病学报,2008,24(1):45-49.FANG H,CHEN C Z,ZHANG X J,et al.Examination of the pathogenic Aeromonas veronii isolated from crab Eriocheir sinensis[J].Chinese Journal of Zoonoses,2008,24(1):45-49.(in Chinese with English abstract)
[13]杨泽晓,周亚,任冉阳,等.白缘()维氏气单胞菌的分离鉴定与药敏试验[J].中国畜牧兽医,2012,39(2):92-96.YANG Z X,ZHOU Y,REN R Y,et al.Isolation,identification and drug susceptibility test of Aeromonas veronii from Liobagrus marginatus Giinther[J].Chinese Journal of Veterinary Medicine,2012,39(2):92-96.(in Chinese with English abstract)
[14]李聪,蔡岩,周永灿,等.海南罗非鱼致病性维氏气单胞菌分离鉴定及药敏特性研究[J].水产科学,2015,34(10):640-646.LI C,CAI Y,ZHOU Y C,et al.Isolation,identification and antibiotic sensitivity of Aeromonas veronii from Tilapia cultured in Hainan Province[J].Fisheries Science,2015,34(10):640-646.(in Chinese with English abstract)
[15]吴同垒,单晓枫,孟庆峰,等.维氏气单胞菌研究进展[J].中国兽药杂志,2011,45(7):41-44.WU T L,SHAN X F,MENG Q F,et al.Advances in Aeromonas veronii[J].Chinese Journal of Veterinary Medicine,2011,45(7):41-44.(in Chinese with English abstract)
[16]ZHU M,WANG X R,LI J,et al.Identification and virulence properties of Aeromonas veronii bv.sobria isolates causing an ulcerative syndrome of loach Misgurnus anguillicaudatus[J].Journal of Fish Diseases,2016,39(6):777-781.
[17]高正勇,曾令兵,孟彦,等.患病大鲵中弗氏柠檬酸杆菌的分离与鉴定[J].微生物学报,2012,52(2):169-176.GAO Z Y,ZENG L B,MENG Y,et al.Isolation and identification of Citrobacter freundii from diseased giant salamander,Andrias davidianus[J].Acta Microbiologica Sinica,2012,52(2):169-176.(in Chinese with English abstract)
[18]周勇,曾令兵,李瑞伟,等.患病黄额闭壳龟中产酸克雷伯菌的分离与鉴定[J].淡水渔业,2012,42(3):38-43.ZHOU Y,ZENG L B,LI R W,et al.Isolation and identification of Klebsiella oxytoca from diseased Cuora galbinifrons[J].Freshwater Fisheries,2012,42(3):38-43.(in Chinese with English abstract)
[19]安伟,肖雨,高晓华,等.鳜源致病性荧光假单胞菌的分离与鉴定[J].动物学杂志,2014,49(5):760-765.AN W,XIAO Y,GAO X H,et al.Isolation and identification of pathogenic Pseudomonas fluorescens from Siniperca chuatsi[J].Chinese Journal of Zoology,2014,49(5):760-765.(in Chinese with English abstract)
[20]吴会桃,蔡芷荷,吴清平,等.细菌鉴定系统的应用研究进展[J].中国卫生检验杂志,2010,20(9):2381-2384.WU H T,CAI Z H,WU Q P,et al.Advances in the application of bacterial identification system[J].Chinese Journal of Health Laboratory Technology,2010,20(9):2381-2384.(in Chinese)