大鲵源杀鲑气单胞菌的分离鉴定及药敏试验
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摘要
为了查明2016年3月份河南省洛阳市某大鲵(Andrias davidianus)养殖厂发生大规模疫病的原因,试验采用平板划线法从患病大鲵中分离细菌,结合革兰氏染色镜检、透射电镜观察、16S rDNA序列分析及遗传进化树构建对其进行鉴定,并对代表株进行生长曲线的测定及人工感染试验以鉴定其致病性,最后进行药敏试验测定水产常用药物的抑菌效果。结果表明:从大鲵组织中共分离出5株细菌,编号为HN1～HN5,均为革兰氏阴性短杆菌,且有一根鞭毛;5株细菌与杀鲑气单胞菌(Aeromonas salmonicida)序列的相似度高达99%,为同种菌株,鉴定其均为杀鲑气单胞菌;代表株HN3菌株与杀鲑气单胞菌LMG 22214株(NR_115351.1)同源性最高; HN3菌株在培养6 h后进入对数生长期,42 h后进入稳定期;HN3菌株为大鲵的病原菌,且对甲砜霉素和恩诺沙星极度敏感,对氟苯尼考、复方新诺明、多西环素高度敏感,对磺胺二甲氧嘧啶钠等4种药物中度敏感,对磺胺间甲氧嘧啶钠等3种药物有耐药性。
In March 2016, a large scale of Andrias davidianus got disease in a fishery in Luoyang city. To find out the reason, bacteria strains were isolated from diseased Andrias davidianus by streak plate method, followed by identification of them by gram stain examination, transmission electron microscopic observation, analysis of 16 S rDNA sequence and phylogenetic tree construction. Strain HN3 was chosen to test the growth curve, pathogenicity and susceptibility to common drugs. Results showed that All isolates,HN1～HN5, were Aeromonas salmonicida which were gram-negative rod bacterium. Aeromonas salmonicida HN3 strain came into logarithm growth period at 6 h post inoculation and stabilization period at 42 h post inoculation. HN3 strain was pathogenic bacteria and extremely sensitive to propacin and enrofloxacin, highly sensitive to florfenicol, sulfametheoxazole and doxycycline, and moderately sensitive to 4 drugs including sulfadimethoxine sodium, but was resistant to 3 drugs including sulfamonomethoxine sodiun. The test lays an academic foundation for the prevention and control of bacterial disease of Andrias davidianus.
In March 2016, a large scale of Andrias davidianus got disease in a fishery in Luoyang city. To find out the reason, bacteria strains were isolated from diseased Andrias davidianus by streak plate method, followed by identification of them by gram stain examination, transmission electron microscopic observation, analysis of 16 S rDNA sequence and phylogenetic tree construction. Strain HN3 was chosen to test the growth curve, pathogenicity and susceptibility to common drugs. Results showed that All isolates,HN1～HN5, were Aeromonas salmonicida which were gram-negative rod bacterium. Aeromonas salmonicida HN3 strain came into logarithm growth period at 6 h post inoculation and stabilization period at 42 h post inoculation. HN3 strain was pathogenic bacteria and extremely sensitive to propacin and enrofloxacin, highly sensitive to florfenicol, sulfametheoxazole and doxycycline, and moderately sensitive to 4 drugs including sulfadimethoxine sodium, but was resistant to 3 drugs including sulfamonomethoxine sodiun. The test lays an academic foundation for the prevention and control of bacterial disease of Andrias davidianus.
引文
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[11] 李绍戊,王荻,连浩淼,等.大西洋鲑杀鲑气单胞菌无色亚种的分离鉴定和致病性研究[J].水生生物学报,2015,39(1):234-240.
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[15] 杨移斌,胥宁,董靖,等.斑点叉尾■ 源杀鲑气单胞菌无色亚种分离鉴定及药敏特性[J].水生生物学报,2017,41(4):787-792.
[16] 杨嘉龙,周丽,邢婧,等.养殖刺参溃疡病杀鲑气单胞菌的分离、致病性及胞外产物特性分析[J].中国水产科学,2007,14(6):981-989.
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[18] 周冬仁,罗毅志,杭小英,等.1株乌鳢源杀鲑气单胞菌杀鲑亚种的分离与鉴定[J].海洋湖沼通报,2015(3):64-70.
[19] FAISAL M,EISSA A E,ELSAYED E E.Isolation of Aeromonas Salomonicida from sealamprey (Petromyzonmarinus) with furuncle-like lesions in lakeontario[J].J Wildlife Dis,2007,43(4):618-622.
[20] 刘宁,时晓,杜迎春,等.患病细鳞鱼杀鲑气单胞菌的分离与鉴定[J].淡水渔业,2015(1):88-92.
[21] 朱成科,周晓扬,张其中.南方鱿幼鱼细菌性败血症病原与组织病理[J].中国水产科学,2011,18(2):360-370.
[22] 陶娅,吴亚光,孙满山,等.猴源猪丹毒杆菌的分离与生物学特性研究[J].黑龙江畜牧兽医,2018(07):130-132.
[23] 高桂生,吉志新,张艳英,等.罗非鱼链球菌的分离鉴定及中草药药敏试验[J].黑龙江畜牧兽医,2016(08下):187-189.
[24] 汪沐,周凤建,张天虎,等.黄颡鱼温和气单胞菌的分离鉴定与中草药药敏试验[J].科学养鱼,2017(6):63-64.
[25] 陈晓利,彭彬,占爱思,等.黄鳝源嗜水气单胞菌的体外中药药敏试验[J].淡水渔业,2014,44(2):43-46.
[2] 杨星,曾令兵,李瑞伟,等.患病大鲵类志贺邻单细胞菌的分离与鉴定[J].水生态学杂志,2014,35(1):91-95.
[3] 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.
[4] 杨星,刘文枝,肖汉兵,等.嗜水气单胞菌灭活疫苗免疫后大鲵外周血免疫指标的变化[J].中国水产科学,2014,21(30):621-628.
[5] 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.
[6] 李瑞伟,曾令兵,张辉,等.患病大鲵摩氏摩根菌的分离与鉴定[J].畜牧与兽医,2014,46(1):28-32.
[7] 敖弟书,吴中明,王玉,等.迟钝爱华德菌感染大鲵的研究[J].四川动物,2010,29(3):411-414.
[8] 王海娟,王利.鲤鱼杀鲑气单胞菌的分离鉴定及耐药性分析[J].中国畜牧兽医,2015,42(1):192-196.
[9] 王家祯,耿昕颖,董文龙,等.3株杀鲑气单胞菌的分离鉴定及药敏试验[J].中国兽医杂志,2017,53(2):78-80.
[10] 丁雷,岳永生,宋憬愚,等.虹鳟皮肤溃烂病的病原菌研究[J].淡水渔业,2002,32(3):28-30.
[11] 李绍戊,王荻,连浩淼,等.大西洋鲑杀鲑气单胞菌无色亚种的分离鉴定和致病性研究[J].水生生物学报,2015,39(1):234-240.
[12] 王高学,黄增荣.白斑狗鱼“败血症”病原及组织病理研究[J].水产学报,2006,30(3):383-387.
[13] LAGO E P,FARTO R.Virulence factors of Aeromonas salmonicida subsp.salmonicida strains associated with infections in turbot Psetta maxima[J].Dis Aquat Organ,2012,99(2):145-151.
[14] HAN J E,KIM J H,SHIN S P,et al.Draft genome sequence of AeromogensAs03,a typical strain isolated from Crucian carp (Carassiuscarassius) in the republic of Korea[J].Genome Announc,2013,1(5):e00791.
[15] 杨移斌,胥宁,董靖,等.斑点叉尾■ 源杀鲑气单胞菌无色亚种分离鉴定及药敏特性[J].水生生物学报,2017,41(4):787-792.
[16] 杨嘉龙,周丽,邢婧,等.养殖刺参溃疡病杀鲑气单胞菌的分离、致病性及胞外产物特性分析[J].中国水产科学,2007,14(6):981-989.
[17] 周红霞,姚俊杰,房文红,等.半滑舌鳎溃疡病原杀鲑气单胞菌的分离鉴定与药敏试验[J].海洋渔业,2017,39(3):322-330.
[18] 周冬仁,罗毅志,杭小英,等.1株乌鳢源杀鲑气单胞菌杀鲑亚种的分离与鉴定[J].海洋湖沼通报,2015(3):64-70.
[19] FAISAL M,EISSA A E,ELSAYED E E.Isolation of Aeromonas Salomonicida from sealamprey (Petromyzonmarinus) with furuncle-like lesions in lakeontario[J].J Wildlife Dis,2007,43(4):618-622.
[20] 刘宁,时晓,杜迎春,等.患病细鳞鱼杀鲑气单胞菌的分离与鉴定[J].淡水渔业,2015(1):88-92.
[21] 朱成科,周晓扬,张其中.南方鱿幼鱼细菌性败血症病原与组织病理[J].中国水产科学,2011,18(2):360-370.
[22] 陶娅,吴亚光,孙满山,等.猴源猪丹毒杆菌的分离与生物学特性研究[J].黑龙江畜牧兽医,2018(07):130-132.
[23] 高桂生,吉志新,张艳英,等.罗非鱼链球菌的分离鉴定及中草药药敏试验[J].黑龙江畜牧兽医,2016(08下):187-189.
[24] 汪沐,周凤建,张天虎,等.黄颡鱼温和气单胞菌的分离鉴定与中草药药敏试验[J].科学养鱼,2017(6):63-64.
[25] 陈晓利,彭彬,占爱思,等.黄鳝源嗜水气单胞菌的体外中药药敏试验[J].淡水渔业,2014,44(2):43-46.
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