罗氏沼虾致病性溶藻弧菌的鉴定及药敏分析
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摘要
自广东省湛江市某养殖场患病的罗氏沼虾幼虾匀浆液内分离到一株优势菌QY170324,通过细菌微量生化鉴定管与16SrDNA序列分析对该株优势菌进行鉴定,并进行药敏特性分析和人工感染试验。结果显示,该菌嗜盐性,兼性厌氧,为革兰氏阴性弧菌,能利用蔗糖、甘露糖,赖氨酸脱羧酶、V-P试验均为阳性,在含3%、6%、8%、10%NaCl胨水中均能生长;根据其菌落形态观察、生理生化特性测定,结合《伯杰氏细菌鉴定手册》,鉴定该菌为溶藻弧菌。通过PCR扩增该菌的16SrDNA基因,经过测序及BLAST比对,菌侏OY170324与溶藻弧菌的同源性高达99%;经人工回归感染试验,该菌有较强的致病力。分离的溶藻弧菌QY170324对庆大霉素、四环素、哌拉西林、氧氟沙星、头孢他啶、复方新诺明等抗生素高度敏感,对头孢曲松、卡那霉素、新霉素、米诺环素、麦迪霉素、头孢唑林、多黏菌素B中度敏感,对克林霉素、羧苄西林、头孢氨苄耐药。
A dominant pathogenic bacterium strain(QY170324)was isolated from juvenile giant freshwater prawn Macrobrachium rosenbergii in a farm in Zhanjiang City,Guangdong Province.The pathogen was identified by bacterial microbiological identification tubes and 16 SrDNA sequence analysis,and antibiotic susceptibility test analysis and artificial infection test were performed.The results showed that this pathogen was a haliphagia,facultative anaerobic and gram-negative bacterium.The lysine decarboxylase and VP test of this pathogen were positive,utilized sucrose and mannose and grew in 3%,6%,8% and10% NaCl peptone water.According to colony morphology,physiological and biochemical characteristics,this pathogen was identified as Vibrio alginolyticus in combination with the " Berger′s Bacteria Identification Manual",with 99% homologous to V.alginolyticus based on 16 SrDNA sequence.The artificial infection test showed that this pathogen was strongly virulent to giant freshwater prawn.Strain QY170324 was highly susceptible to gentamicin,tetracycline,piperacillin,ceftazidime,complex sinomin and ofloxacin,intermediately susceptible to cefatriaxone,kanamycin,neomycin,minocycline,maddie,cephazoline and polymyxin B,and resistant to clindamycin,oxacillin,and cephalexin.
A dominant pathogenic bacterium strain(QY170324)was isolated from juvenile giant freshwater prawn Macrobrachium rosenbergii in a farm in Zhanjiang City,Guangdong Province.The pathogen was identified by bacterial microbiological identification tubes and 16 SrDNA sequence analysis,and antibiotic susceptibility test analysis and artificial infection test were performed.The results showed that this pathogen was a haliphagia,facultative anaerobic and gram-negative bacterium.The lysine decarboxylase and VP test of this pathogen were positive,utilized sucrose and mannose and grew in 3%,6%,8% and10% NaCl peptone water.According to colony morphology,physiological and biochemical characteristics,this pathogen was identified as Vibrio alginolyticus in combination with the " Berger′s Bacteria Identification Manual",with 99% homologous to V.alginolyticus based on 16 SrDNA sequence.The artificial infection test showed that this pathogen was strongly virulent to giant freshwater prawn.Strain QY170324 was highly susceptible to gentamicin,tetracycline,piperacillin,ceftazidime,complex sinomin and ofloxacin,intermediately susceptible to cefatriaxone,kanamycin,neomycin,minocycline,maddie,cephazoline and polymyxin B,and resistant to clindamycin,oxacillin,and cephalexin.
引文
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[16]刘淇,李海燕,王群,等.梭子蟹牙膏病病原菌—溶藻弧菌的鉴定及其系统发育分析[J].海洋水产研究,2007,28(4):9-13.
[17]李正义,贾俊涛,曹际娟,等.一株分离自冻墨鱼的溶藻弧菌的鉴定及其低温下细胞脂肪酸的变化[J].水产科学,2014,33(5):317-321.
[18]徐晓丽,叶红梅,侯纯强,等.半滑舌鳎弧菌病病原的分离鉴定[J].中国兽医杂志,2017(8):86-90.
[19]程俊茗,万明月,周晋扬,等.鲫源迟缓爱德华氏菌的分离鉴定及其毒力基因的检测[J].微生物学通报,2017,44(10):2380-2390.
[20]Beleneva I A.Physiological and biochemical characteristics of the halophilic bacteria Vibrio parahaemolyticus and V.alginolyticus isolated from marine invertebrates of Peter the Great Bay,Sea of Japan[J].Microbiology,2004,30(2):96-100.
[21]刁菁,杨秀生,叶海斌,等.一种水产迟钝爱德华氏菌快速药敏检测方法的研究[J].水产科学,2014,33(1):22-28.
[2]朱锦裕,卜弘毅,胡冲冲,等,泼洒糖蜜对池塘养殖罗氏沼虾生长和水质的影响[J].水产科学,2017,36(2):202-206.
[3]Alambra J R,Alenton R R R,Gulpeo P C R,et al.Immunomodulatory effects of turmeric,Curcuma longa(Magnoliophyta,Zingiberaceae)on Macrobrachium[J].Aquaculture,Aquarium,Conservation&Legislation,2012(5):13-17.
[4]刘海春,董学洪,刘志国,等.罗氏沼虾健康养殖水质调控技术[J].江苏农业科学,2011,39(10):338-341.
[5]Kennedy B.Bacterial flora associated with the giant freshwater prawn Macrobrachium rosenbergii,in the hatchery system[J].Aquaculture,2006,261(4):1156-1167.
[6]Lee S W,Najiah M.Multiple antibiotic resistance and heavy metal resistance profile of bacteria isolated from giant freshwater prawn(Macrobrachium rosenbergii)hatchery[J].Science Direct,2009,8(6):740-745.
[7]Wikler M A.M100-S18,Performance standards for antimicrobial susceptibility testing:Eighteenth Informationgal Supplement[S].USA:Clinical and Laboratory Standards Institute,2008.
[8]滕勇勇,王琪,吴雷,等.致病性弧菌的生物学特性和致病因子研究进展[J].热带医学杂志,2014,14(10):1396-1399.
[9]陈强,鄢庆枇,马甡,等.溶藻弧菌致病性研究进展[J].海洋科学,2006,30(8):83-89.
[10]林永添.紫贻贝(Mytilus edulis Linnaeus)大量死亡原因的初步研究[J].现代渔业信息,2006,21(7):23-26.
[11]韩风杰,徐文娜,刘敏,等.魁蚶溶藻弧菌分离鉴定与药敏试验[J].山东畜牧兽医,2016,37(5):13-14.
[12]桑本红,徐晓津,毛勇,等.大黄鱼溶藻弧菌病组织中巨噬细胞移动抑制因子的检测及意义[J].海洋学报,2014,36(8):111-116.
[13]赖迎迢,陶家发,孙承文,等.鱼源溶藻弧菌生物学特性和病理组织学观察[J].微生物学报,2014,54(11):1378-1384.
[14]陈寅儿,金珊,王国良,等.鲈鱼溶藻弧菌病的血液生理生化指标研究[J].台湾海峡,2005,24(1):104-108.
[15]罗词兴,黄旭雄,李桑,等.溶藻弧菌感染后凡纳滨对虾鳃组织免疫相关基因的表达[J].中国水产科学,2014,21(1):189-196.
[16]刘淇,李海燕,王群,等.梭子蟹牙膏病病原菌—溶藻弧菌的鉴定及其系统发育分析[J].海洋水产研究,2007,28(4):9-13.
[17]李正义,贾俊涛,曹际娟,等.一株分离自冻墨鱼的溶藻弧菌的鉴定及其低温下细胞脂肪酸的变化[J].水产科学,2014,33(5):317-321.
[18]徐晓丽,叶红梅,侯纯强,等.半滑舌鳎弧菌病病原的分离鉴定[J].中国兽医杂志,2017(8):86-90.
[19]程俊茗,万明月,周晋扬,等.鲫源迟缓爱德华氏菌的分离鉴定及其毒力基因的检测[J].微生物学通报,2017,44(10):2380-2390.
[20]Beleneva I A.Physiological and biochemical characteristics of the halophilic bacteria Vibrio parahaemolyticus and V.alginolyticus isolated from marine invertebrates of Peter the Great Bay,Sea of Japan[J].Microbiology,2004,30(2):96-100.
[21]刁菁,杨秀生,叶海斌,等.一种水产迟钝爱德华氏菌快速药敏检测方法的研究[J].水产科学,2014,33(1):22-28.