鲟源荧光假单胞菌的分离鉴定及药敏特性分析
|
摘要
从患病鲟体内分离到1株细菌,通过形态学观察、生理生化特性鉴定、16S rRNA基因序列分析及系统进化树构建等方法对分离菌种属进行确定,采用纸片扩散法对分离菌进行药物敏感性分析。结果显示:分离菌为革兰阴性杆菌;可分解葡萄糖和木糖,氧化酶和鸟氨酸脱羧酶试验阳性;16S rRNA基因测序分析与荧光假单胞菌(Pseudomonas fluorescens)同源性大于99%,在系统发育树上与荧光假单胞菌聚为一枝;药敏结果显示,分离菌对氧氟沙星、多西环素、复方新诺明等敏感,对氟苯尼考、卡那霉素、阿莫西林等耐药;人工感染试验结果显示,该菌对小鼠有致病力。本研究为贵州地区鲟细菌性疾病预防及合理用药提供依据。
The present study was to identify the bacteria isolated from sturgeon and to analyze their antibiotic sensitivity. A strain of bacteria, GZZY2018, was isolated from the diseased mites, and the isolates were identified by morphological observation, physiological and biochemical characterization, 16 S rRNA gene sequence analysis and phylogenetic tree construction, analysis of drug sensitivity of isolates by antibiotic sensitivity test. The results showed that GZZY2018 was Gram-negative bacilli, which decomposes glucose and xylose, and their oxidase and ornithine decarboxylase were positive. Sequencing analysis of the 16 S rRNA gene showed its homology with Pseudomonas fluorescens was over 99%, and the gene clustered with P. fluorescens in the phylogenetic tree. The antibiotic sensitivity test showed that the strain GZZY2018 was sensitive to ofloxacin, doxycycline and compound neotamine, but was resistant to florfenicol, kanamycin and amoxicillin. The artificial infection test showed that the bacteria was pathogenic in mice.
The present study was to identify the bacteria isolated from sturgeon and to analyze their antibiotic sensitivity. A strain of bacteria, GZZY2018, was isolated from the diseased mites, and the isolates were identified by morphological observation, physiological and biochemical characterization, 16 S rRNA gene sequence analysis and phylogenetic tree construction, analysis of drug sensitivity of isolates by antibiotic sensitivity test. The results showed that GZZY2018 was Gram-negative bacilli, which decomposes glucose and xylose, and their oxidase and ornithine decarboxylase were positive. Sequencing analysis of the 16 S rRNA gene showed its homology with Pseudomonas fluorescens was over 99%, and the gene clustered with P. fluorescens in the phylogenetic tree. The antibiotic sensitivity test showed that the strain GZZY2018 was sensitive to ofloxacin, doxycycline and compound neotamine, but was resistant to florfenicol, kanamycin and amoxicillin. The artificial infection test showed that the bacteria was pathogenic in mice.
引文
[1]刘晓勇,李鸶鸶,赵明军.中国鲟鱼养殖产业发展战略思考[J].中国渔业经济,2013,31(06):69-76.
[2]Zee J,Patterson S,Gagnon D,et al.Adverse health effects and histological changes in white sturgeon(Acipenser transmontanus)exposed to dietary selenomethionine[J].Environ Toxicol Chem,2016,35(7):1741-50.
[3]Poleksic V,Lenhardt M,Jaric I,et al.Liver,gills,and skin histopathology and heavy metal content of the danube sterlet(Acipenser ruthenuslinnaeus,1758)[J].Environ Toxicol Chem,2010,29(3):515-21.
[4]Ostazewska T,Chojnacki M,Kamasewski M,et al.Histopathological effects of silver and copper nanoparticles on the epidermis,gills,and liver of Siberian sturgeon[J].Environ Sci Pollut Res Int,2016,23(2):1621-33.
[5]刘晓勇,齐茜,王国峰,等.养殖密度对饲料驯化期西伯利亚鲟生长的影响[J].淡水渔业,2012,42(05):79-82.
[6]张溢卓,赵明军.我国鲟鱼产业现状分析及其对策建议[J].饲料究,2017(14):33-37+43.
[6]宫民,刘丹阳.鲟鱼营养价值研究进展[J].黑龙江水产,2018(04):10-11.
[7]张溢卓,赵明军.我国鲟鱼产业现状分析及其对策建议[J].饲料研究,2017(14):33-37+43.
[8]王世权.鲟鱼常见病的治疗[J].中国水产,1999(10):32-33.
[9]陈细华,李创举,杨长庚,等.中国鲟鱼产业技术研发现状与展望[J].淡水渔业,2017,47(06):108-112.
[10]张艳珍,张晓雁,王彦鹏.养殖中华鲟四种天然饵料的细菌分布及灭菌效果[J].淡水渔业,2014,44(03):90-94.
[11]王有基,胡梦红.人工养殖鲟鱼的疾病与防治[J].北京农业,2004(12):26-27.
[12]王亚霞,苏振杰,任凤秋.K-B纸片琼脂扩散法检测细菌药物敏感试验的影响因素分析[J].中国医药指南,2009,7(05):41-42.
[13]Swain P,Behura A,Dash S,et al.Serum antibody response of Indian major carp,Labeo rohita to three species of pathogenic bacteria;Aeromonas hydrophila,Edwardsiella tarda and Pseudomonas fluorescens[J].Vet Immunol Immunopathol,2007,117(1-2):137-141.
[14]Shah S Q A,Srum H.Genetic localization of a Tet R-like transcriptional regulator gene in Pseudomonas fluorescens isolated from farmed fish[J].J Appl Genet,2014,55(4):541-544.doi:10.
[15]邓显文,谢芝勋,刘加波,等.罗非鱼荧光假单胞菌的分离鉴定[J].广西农业科学,2010,41(06):612-615.
[16]杜宗君,黄小丽,邓永强,等.人工养殖大鲵病原荧光假单胞菌的分离鉴定[J].四川农业大学学报,2011,29(01):103-107.
[17]沈晓静,胡秀彩,兰云,等.锦鲤荧光假单胞菌的分离鉴定及药敏试验[J].水产科学,2014,33(07):443-446.
[18]李道友,李正友,周路,等.盐酸恩诺沙星防治鲟鱼细菌性鱼病[J].贵州畜牧兽医,2011,35(06):67-68.
[19]Sun YY,Sun L.Pseudomonas fluorescens:iron-responsive proteins and their involvement in host infection.Vet Microbiol[J].2015,176(3-4):309-320.
[20]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(05):760-765.
[21]高桂生,张艳英,吉志新,等.半滑舌鳎致病性荧光假单胞菌的分离鉴定及其感染的病理损伤[J].中国兽医学报,2016,36(07):1145-1150.
[2]Zee J,Patterson S,Gagnon D,et al.Adverse health effects and histological changes in white sturgeon(Acipenser transmontanus)exposed to dietary selenomethionine[J].Environ Toxicol Chem,2016,35(7):1741-50.
[3]Poleksic V,Lenhardt M,Jaric I,et al.Liver,gills,and skin histopathology and heavy metal content of the danube sterlet(Acipenser ruthenuslinnaeus,1758)[J].Environ Toxicol Chem,2010,29(3):515-21.
[4]Ostazewska T,Chojnacki M,Kamasewski M,et al.Histopathological effects of silver and copper nanoparticles on the epidermis,gills,and liver of Siberian sturgeon[J].Environ Sci Pollut Res Int,2016,23(2):1621-33.
[5]刘晓勇,齐茜,王国峰,等.养殖密度对饲料驯化期西伯利亚鲟生长的影响[J].淡水渔业,2012,42(05):79-82.
[6]张溢卓,赵明军.我国鲟鱼产业现状分析及其对策建议[J].饲料究,2017(14):33-37+43.
[6]宫民,刘丹阳.鲟鱼营养价值研究进展[J].黑龙江水产,2018(04):10-11.
[7]张溢卓,赵明军.我国鲟鱼产业现状分析及其对策建议[J].饲料研究,2017(14):33-37+43.
[8]王世权.鲟鱼常见病的治疗[J].中国水产,1999(10):32-33.
[9]陈细华,李创举,杨长庚,等.中国鲟鱼产业技术研发现状与展望[J].淡水渔业,2017,47(06):108-112.
[10]张艳珍,张晓雁,王彦鹏.养殖中华鲟四种天然饵料的细菌分布及灭菌效果[J].淡水渔业,2014,44(03):90-94.
[11]王有基,胡梦红.人工养殖鲟鱼的疾病与防治[J].北京农业,2004(12):26-27.
[12]王亚霞,苏振杰,任凤秋.K-B纸片琼脂扩散法检测细菌药物敏感试验的影响因素分析[J].中国医药指南,2009,7(05):41-42.
[13]Swain P,Behura A,Dash S,et al.Serum antibody response of Indian major carp,Labeo rohita to three species of pathogenic bacteria;Aeromonas hydrophila,Edwardsiella tarda and Pseudomonas fluorescens[J].Vet Immunol Immunopathol,2007,117(1-2):137-141.
[14]Shah S Q A,Srum H.Genetic localization of a Tet R-like transcriptional regulator gene in Pseudomonas fluorescens isolated from farmed fish[J].J Appl Genet,2014,55(4):541-544.doi:10.
[15]邓显文,谢芝勋,刘加波,等.罗非鱼荧光假单胞菌的分离鉴定[J].广西农业科学,2010,41(06):612-615.
[16]杜宗君,黄小丽,邓永强,等.人工养殖大鲵病原荧光假单胞菌的分离鉴定[J].四川农业大学学报,2011,29(01):103-107.
[17]沈晓静,胡秀彩,兰云,等.锦鲤荧光假单胞菌的分离鉴定及药敏试验[J].水产科学,2014,33(07):443-446.
[18]李道友,李正友,周路,等.盐酸恩诺沙星防治鲟鱼细菌性鱼病[J].贵州畜牧兽医,2011,35(06):67-68.
[19]Sun YY,Sun L.Pseudomonas fluorescens:iron-responsive proteins and their involvement in host infection.Vet Microbiol[J].2015,176(3-4):309-320.
[20]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(05):760-765.
[21]高桂生,张艳英,吉志新,等.半滑舌鳎致病性荧光假单胞菌的分离鉴定及其感染的病理损伤[J].中国兽医学报,2016,36(07):1145-1150.