鲫鱼维隆气单胞菌和腐败希瓦菌的分离鉴定、致病性和药敏分析
|
摘要
从患病鲫鱼肝、肾、血体内分离到JYV1605101和JYF1605102 2株细菌,人工感染试验和溶血活性测定表明这2株菌对健康鲫鱼具有较强毒力,菌株JYV1605101对鲫鱼LD_(50)为3.31×10~3cfu·g~(-1),对绵羊、小鼠、乌鳢、鲫鱼、鲤鱼、加州鲈等6种动物血细胞呈β溶血;菌株JYF1605102对鲫鱼LD_(50)为1.48×10~2cfu·g~(-1),对乌鳢、鲫鱼、鲤鱼、加州鲈的血细胞呈β溶血,但对绵羊和小鼠血细胞呈α溶血.经形态学观察,生理生化试验及16S rDNA基因序列分析,鉴定菌株JYV1605101为维隆气单胞菌(Aeromonas veronii),菌株JYF1605102为腐败希瓦菌(Shewanella putrefaciens).菌株JYV1605101具有胞外蛋白酶、明胶酶、淀粉酶活性,不具有脲酶、脂酶和卵磷脂酶活性;菌株JYF1605102具有胞外蛋白酶、明胶酶活性,不具有脲酶、脂酶、淀粉酶和卵磷脂酶活性.药敏试验表明,2株菌都对其中的18种药物较敏感,根据渔业用药规定和常用药情况,氟苯尼考、四环素、米诺环素、强力霉素可作为鲫鱼维隆气单胞菌和腐败希瓦菌感染的候选控制药物.
Two bacterial strains were isolated from sick crucian carp,Carassiu sauratus,JYV1605101 and JYV1605102.The artificial infection test and hemolytic tests suggested both strains were highly toxic.The LD_(50)of strain JYV1605101 to crucian carp was 3.90×10~6cfu·mL~(-1).Strain JYV1605101 isβ-hemolytic to blood of Ovis aries,Mus musculus,Channa argus,Carassius auratus,Cyprinus carpio and Micropterus salmoides.The LD_(50) of strain JYF1605102 to crucian carp was 2.36×10~5cfu·mL~(-1).Strain JYV1605102 isβ-hemolytic to blood of C.argus,C.auratus,C.carpio and M.salmoides andα-hemolytic to blood of O.aries and M.musculus.The morphology observation,biochemical characteristic and 16s rDNA sequencing analysis showed that strain JYV1605101 was A.veronii and strain JYV1605102 was S.putrefaciens.Strain JYV1605101 produces the extracellular enzyme activity of protease,gelatinase and amylase,rather than the activity of urease,lipase and lecithinase.Strain JYF1605102 has the extracellular enzyme activity of protease and gelatinase,rather than the activity of amylase,urease,lipase and lecithinase.The result of drug sensitivity test demonstrated that both strains are relatively sensitive to18 types of drugs.According to the regulation of fishery medication and usage of routine drugs,it is suggested that florfenicol,tetracycline,minocycline and doxycycline could be considered as the candidate treatment to the crucian carp diseases caused by A.veronii and S.putrefaciens.
Two bacterial strains were isolated from sick crucian carp,Carassiu sauratus,JYV1605101 and JYV1605102.The artificial infection test and hemolytic tests suggested both strains were highly toxic.The LD_(50)of strain JYV1605101 to crucian carp was 3.90×10~6cfu·mL~(-1).Strain JYV1605101 isβ-hemolytic to blood of Ovis aries,Mus musculus,Channa argus,Carassius auratus,Cyprinus carpio and Micropterus salmoides.The LD_(50) of strain JYF1605102 to crucian carp was 2.36×10~5cfu·mL~(-1).Strain JYV1605102 isβ-hemolytic to blood of C.argus,C.auratus,C.carpio and M.salmoides andα-hemolytic to blood of O.aries and M.musculus.The morphology observation,biochemical characteristic and 16s rDNA sequencing analysis showed that strain JYV1605101 was A.veronii and strain JYV1605102 was S.putrefaciens.Strain JYV1605101 produces the extracellular enzyme activity of protease,gelatinase and amylase,rather than the activity of urease,lipase and lecithinase.Strain JYF1605102 has the extracellular enzyme activity of protease and gelatinase,rather than the activity of amylase,urease,lipase and lecithinase.The result of drug sensitivity test demonstrated that both strains are relatively sensitive to18 types of drugs.According to the regulation of fishery medication and usage of routine drugs,it is suggested that florfenicol,tetracycline,minocycline and doxycycline could be considered as the candidate treatment to the crucian carp diseases caused by A.veronii and S.putrefaciens.
引文
[1]杨秀雄.高密度养殖鲫鱼常见疾病防治法[J].吉林水产, 2015(16):81.
[2]韩艳楠,余银春,王鑫毅,等.牛蛙致病变形菌的鉴定及其敏感药物筛选[J].微生物学通报,2016,43(6):1280-1287.
[3]巴翠玉,赵福广,张培军,等.鲫鱼戴氏西地西菌的分离与鉴定[J].中国兽医杂志, 2016, 52(5):107-109.
[4]秦蕾,张晓君,毕可然.一种新的异育银鲫病原—–腐败希瓦菌[J].微生物学报, 2012, 52(5):558-565.
[5]王利,魏勇,龙钟明,等.鲫鱼弗氏柠檬酸杆菌的耐药表型及耐药基因检测[J].黑龙江畜牧兽医,2012(8):14-17.
[6]陆文浩,杨家新,陈辉,等.异育银鲫类志贺邻单胞菌的鉴定[J].淡水渔业, 2009, 39(2):48-53.
[7]陆文浩,陈辉,王习达,等.异育银鲫致病性鲍曼不动杆菌的鉴定和系统发育分析[J].中国兽医科学,2009,39(4):303-309.
[8]林秀秀.患鲤疱疹Ⅱ型病毒病异育银鲫(Carassius auratusgibelio)组织和血液病理的研究[D].苏州:苏州大学, 2016.
[9]巴翠玉,张培军,赵福广,等.异育银鲫呼肠孤病毒的分离与鉴定[J].华南农业大学学报, 2017(2):22-26.
[10] Lu H D, Zhu G Z, Fan L P, et al. Etiology and pathology of epidermal papillomas in Allogynogenetic crucian carp(Carassiusauratusgibelio♀xCyprinuscarpiovar.singuonensis♂)[J]. Diseases of Aquatic Organisms, 2009,83(1):77-84.
[11]王鑫毅,韩艳楠,金珊,等.乌鳢产吲哚金黄杆菌的鉴定及其胞外产物特性分析[J].水生生物学报,2016,40(3):641-646.
[12] SatomiM,VogelBF,GramL,etal.Shewanella hafniensissp.nov.andShewanellamorhuaesp.nov.isolatedfrommarinefishoftheBalticSea[J].InternationalJournalofSystematicandEvolutionary Microbiology, 2006, 56(1):243-249.
[13] Ziemke F, H?fle M G, Lalucat J, et al. Reclassification of ShewanellaputrefaciensOwen’sgenomicgroupIIas Shewanellabalticasp.nov.[J].InternationalJournalof Systematic and Evolutionary Microbiology, 1998, 48(1):179-186.
[14]赵乃昕,苑广盈.医学细菌名称及分类鉴定[M].3版.济南:山东大学出版社, 2013:115-120; 173-179.
[15]房海,陈翠真,张晓君.水产养殖动物病原细菌学[M].北京:中国农业出版社, 2010:389-446.
[16]孙瑞元.简捷实用的半数致死量综合计算法[J].药学学报, 1963, 10(2):65-74.
[17]徐华.红斑狼疮细胞检查方法的改进[J].四川省卫生管理干部学院学报, 1996, 15(3):182.
[18]吴同垒,单晓枫,孟庆峰,等.维隆气单胞菌研究进展[J].中国兽药杂志, 2011, 45(7):41-44.
[19]何涛,邹升,龚亮,等.草鱼病原菌Av X005的分离鉴定及其拮抗菌筛选研究[J].水产科学,2018,37(1):15-23.
[20]胡秀彩,孙金辉,孙敬锋,等.患病蝶尾金鱼维隆气单胞菌温和生物变种的分离鉴定[J].水生态学杂志,2018, 39(1):105-110.
[21]贾俊鹏,康元环,张冬星,等.锦鲫致病性维隆气单胞菌的分离鉴定及耐药性分析[J].中国预防兽医学报,2018, 40(1):19-23.
[22]赵宇亮,吕梦溪,苏欣,等.中华鳖病原维隆气单胞菌的分子鉴定及分子分类学研究[J].河北师范大学学报(自然科学版), 2011, 35(4):417-422.
[23]房海,陈翠珍,张晓君,等.中华绒螯蟹病原维隆气单胞菌的检验[J].中国人兽共患病学报,2008,24(1):45-49.
[24]葛铮.不同淡水鱼类维隆气单胞菌的分离鉴定及耐药性分析[D].长春:吉林农业大学, 2015.
[25]周光,谢骏,周群兰,等.团头鲂养殖池塘维隆气单胞菌的致病性耐药性及在不同生态位间的差异[J].江苏农业科学, 2012, 40(5):192-196.
[26]王旭,颜其贵,雷燕,等.中国大鲵腐皮病病原菌的分离与鉴定[J].中国人兽共患病学报,2010,26(10):944-948.
[27]王庆丽.养殖大黄鱼和海水鲈鱼冷藏过程中特定腐败菌的鉴别[D].杭州:浙江工商大学, 2012.
[28]成沛.东方欧鳊溃烂病病原菌的分离鉴定及流行病学研究[D].乌鲁木齐:新疆农业大学, 2013.
[29]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].青岛:中国海洋大学, 2008.
[30]徐欣,陈军昌.五种中草药对河蟹腐败假单胞菌的抑菌作用[J].水产科学, 2002, 21(2):24-27.
[31]樊汶樵,周莉,廖冰洁,等.感染棘腹蛙的腐败希瓦菌的分离与鉴定[J].黑龙江畜牧兽医,2015(15):166-168;299.
[32] KozinskaA,PekalaA.FirstisolationofShewanella putrefaciensfromfreshwaterfish—apotentialnew pathogenoffish[J].BulletinEuropeanAssociationof Fish Pathologists, 2004, 24:189-193.
[33] SaeedMO,AlamoudiMM,Al-HarbiAH.A Pseudomonas associated with disease in cultured rabbitfishSiganusrivulatusintheRedSea[J].Diseasesof Aquatic Organisms, 1987, 3:177-180.
[34] Korun J, Akgün-Dar K, Yazici M. Isolation of Shewanella putrefaciensfromculturedEuropeanseabass,(Dicentrarchus labrax)in Turkey[J]. Revue de Médecine Vétérinaire, 2009, 160:532-536.
[35]黎炯,叶星,卢迈新,等.罗非鱼维隆气单胞菌的分离鉴定和药敏试验[J].水生态学杂志,2011,32(3):132-136.
[36]贾春红,李淑芳,张继东.3株方斑东风螺病原希瓦菌对35种常用抗菌药物的敏感性试验[J].江西水产科技,2013(1):17-19.
[2]韩艳楠,余银春,王鑫毅,等.牛蛙致病变形菌的鉴定及其敏感药物筛选[J].微生物学通报,2016,43(6):1280-1287.
[3]巴翠玉,赵福广,张培军,等.鲫鱼戴氏西地西菌的分离与鉴定[J].中国兽医杂志, 2016, 52(5):107-109.
[4]秦蕾,张晓君,毕可然.一种新的异育银鲫病原—–腐败希瓦菌[J].微生物学报, 2012, 52(5):558-565.
[5]王利,魏勇,龙钟明,等.鲫鱼弗氏柠檬酸杆菌的耐药表型及耐药基因检测[J].黑龙江畜牧兽医,2012(8):14-17.
[6]陆文浩,杨家新,陈辉,等.异育银鲫类志贺邻单胞菌的鉴定[J].淡水渔业, 2009, 39(2):48-53.
[7]陆文浩,陈辉,王习达,等.异育银鲫致病性鲍曼不动杆菌的鉴定和系统发育分析[J].中国兽医科学,2009,39(4):303-309.
[8]林秀秀.患鲤疱疹Ⅱ型病毒病异育银鲫(Carassius auratusgibelio)组织和血液病理的研究[D].苏州:苏州大学, 2016.
[9]巴翠玉,张培军,赵福广,等.异育银鲫呼肠孤病毒的分离与鉴定[J].华南农业大学学报, 2017(2):22-26.
[10] Lu H D, Zhu G Z, Fan L P, et al. Etiology and pathology of epidermal papillomas in Allogynogenetic crucian carp(Carassiusauratusgibelio♀xCyprinuscarpiovar.singuonensis♂)[J]. Diseases of Aquatic Organisms, 2009,83(1):77-84.
[11]王鑫毅,韩艳楠,金珊,等.乌鳢产吲哚金黄杆菌的鉴定及其胞外产物特性分析[J].水生生物学报,2016,40(3):641-646.
[12] SatomiM,VogelBF,GramL,etal.Shewanella hafniensissp.nov.andShewanellamorhuaesp.nov.isolatedfrommarinefishoftheBalticSea[J].InternationalJournalofSystematicandEvolutionary Microbiology, 2006, 56(1):243-249.
[13] Ziemke F, H?fle M G, Lalucat J, et al. Reclassification of ShewanellaputrefaciensOwen’sgenomicgroupIIas Shewanellabalticasp.nov.[J].InternationalJournalof Systematic and Evolutionary Microbiology, 1998, 48(1):179-186.
[14]赵乃昕,苑广盈.医学细菌名称及分类鉴定[M].3版.济南:山东大学出版社, 2013:115-120; 173-179.
[15]房海,陈翠真,张晓君.水产养殖动物病原细菌学[M].北京:中国农业出版社, 2010:389-446.
[16]孙瑞元.简捷实用的半数致死量综合计算法[J].药学学报, 1963, 10(2):65-74.
[17]徐华.红斑狼疮细胞检查方法的改进[J].四川省卫生管理干部学院学报, 1996, 15(3):182.
[18]吴同垒,单晓枫,孟庆峰,等.维隆气单胞菌研究进展[J].中国兽药杂志, 2011, 45(7):41-44.
[19]何涛,邹升,龚亮,等.草鱼病原菌Av X005的分离鉴定及其拮抗菌筛选研究[J].水产科学,2018,37(1):15-23.
[20]胡秀彩,孙金辉,孙敬锋,等.患病蝶尾金鱼维隆气单胞菌温和生物变种的分离鉴定[J].水生态学杂志,2018, 39(1):105-110.
[21]贾俊鹏,康元环,张冬星,等.锦鲫致病性维隆气单胞菌的分离鉴定及耐药性分析[J].中国预防兽医学报,2018, 40(1):19-23.
[22]赵宇亮,吕梦溪,苏欣,等.中华鳖病原维隆气单胞菌的分子鉴定及分子分类学研究[J].河北师范大学学报(自然科学版), 2011, 35(4):417-422.
[23]房海,陈翠珍,张晓君,等.中华绒螯蟹病原维隆气单胞菌的检验[J].中国人兽共患病学报,2008,24(1):45-49.
[24]葛铮.不同淡水鱼类维隆气单胞菌的分离鉴定及耐药性分析[D].长春:吉林农业大学, 2015.
[25]周光,谢骏,周群兰,等.团头鲂养殖池塘维隆气单胞菌的致病性耐药性及在不同生态位间的差异[J].江苏农业科学, 2012, 40(5):192-196.
[26]王旭,颜其贵,雷燕,等.中国大鲵腐皮病病原菌的分离与鉴定[J].中国人兽共患病学报,2010,26(10):944-948.
[27]王庆丽.养殖大黄鱼和海水鲈鱼冷藏过程中特定腐败菌的鉴别[D].杭州:浙江工商大学, 2012.
[28]成沛.东方欧鳊溃烂病病原菌的分离鉴定及流行病学研究[D].乌鲁木齐:新疆农业大学, 2013.
[29]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].青岛:中国海洋大学, 2008.
[30]徐欣,陈军昌.五种中草药对河蟹腐败假单胞菌的抑菌作用[J].水产科学, 2002, 21(2):24-27.
[31]樊汶樵,周莉,廖冰洁,等.感染棘腹蛙的腐败希瓦菌的分离与鉴定[J].黑龙江畜牧兽医,2015(15):166-168;299.
[32] KozinskaA,PekalaA.FirstisolationofShewanella putrefaciensfromfreshwaterfish—apotentialnew pathogenoffish[J].BulletinEuropeanAssociationof Fish Pathologists, 2004, 24:189-193.
[33] SaeedMO,AlamoudiMM,Al-HarbiAH.A Pseudomonas associated with disease in cultured rabbitfishSiganusrivulatusintheRedSea[J].Diseasesof Aquatic Organisms, 1987, 3:177-180.
[34] Korun J, Akgün-Dar K, Yazici M. Isolation of Shewanella putrefaciensfromculturedEuropeanseabass,(Dicentrarchus labrax)in Turkey[J]. Revue de Médecine Vétérinaire, 2009, 160:532-536.
[35]黎炯,叶星,卢迈新,等.罗非鱼维隆气单胞菌的分离鉴定和药敏试验[J].水生态学杂志,2011,32(3):132-136.
[36]贾春红,李淑芳,张继东.3株方斑东风螺病原希瓦菌对35种常用抗菌药物的敏感性试验[J].江西水产科技,2013(1):17-19.