异育银鲫源嗜水气单胞菌对磺胺类耐药性分析
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摘要
为了解异育银鲫源嗜水气单胞菌对磺胺类药物的敏感性和磺胺类耐药基因的携带情况。采用纸片扩散法和微量肉汤稀释法测定了36株受试菌株对常见4种磺胺类药物的敏感性;应用PCR法检测菌株染色体DNA和质粒DNA磺胺类Sul1、Sul2和Sul3耐药基因。试验结果表明,试验菌株对磺胺嘧啶、磺胺二甲嘧啶、磺胺异噁唑、磺胺甲噁唑的耐药率分别为83.34%、72.22%、66.66%、69.45%;36株嗜水气单胞菌的染色体和质粒中均检测到Sul1和Sul2基因,Sul1基因在染色体和质粒中的检出率分别为30.56%和25.64%,Sul2基因在染色体和质粒中的检出率分别为83.33%和91.67%,Sul3基因在染色体和质粒中均未检出。异育银鲫源嗜水气单胞菌对磺胺类药物的耐药性不容乐观,其耐药性与Sul2和Sul1基因有较大相关性。
The sensitivity of 36 strains of Aeromonas hydrophilaisolated from allogyogenetic silver crucian carp Carassius auratus gibelio to four common sulfonamides was studied by disk diffusion method and broth dilution method and the sulfa drug resistance genes about Sul1,Sul2 and Sul3 were detected by a PCR method from chromosome of DNA and plasmid DNA in order to understand the sulfa drug sensitivity and sulfa drug resistance genes.It was found that the test strains had drug resistance rate of 83.34% to sulfadiazine,72.22% to sulfamethazine,and 66.66% to sulfamethoxazole,and 69.45% to sulfamethoxazole.A total of 36 strains of A.hydrophila chromosomes and plasmids were detected in Sul1 and Sul2 genes,with Sul1 gene detection rate of 30.56%in the chromosome and 25.64% in plasmid,Sul2 gene 83.33%in the chromosome and 91.67% in plasmid and without detection Sul3 gene in chromosome and plasmid.It is not optimistic about the sulfa drugs resistance of A.hydrophilaisolated from allogyogenetic silver crucian carp,and its resistance is closely related to genes Sul2 and Sul1.
The sensitivity of 36 strains of Aeromonas hydrophilaisolated from allogyogenetic silver crucian carp Carassius auratus gibelio to four common sulfonamides was studied by disk diffusion method and broth dilution method and the sulfa drug resistance genes about Sul1,Sul2 and Sul3 were detected by a PCR method from chromosome of DNA and plasmid DNA in order to understand the sulfa drug sensitivity and sulfa drug resistance genes.It was found that the test strains had drug resistance rate of 83.34% to sulfadiazine,72.22% to sulfamethazine,and 66.66% to sulfamethoxazole,and 69.45% to sulfamethoxazole.A total of 36 strains of A.hydrophila chromosomes and plasmids were detected in Sul1 and Sul2 genes,with Sul1 gene detection rate of 30.56%in the chromosome and 25.64% in plasmid,Sul2 gene 83.33%in the chromosome and 91.67% in plasmid and without detection Sul3 gene in chromosome and plasmid.It is not optimistic about the sulfa drugs resistance of A.hydrophilaisolated from allogyogenetic silver crucian carp,and its resistance is closely related to genes Sul2 and Sul1.
引文
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[22]赖海梅,刘书亮,邹立扣,等.肉鸡屠宰场多重耐药沙门氏菌Ⅰ类整合子与磺胺类耐药基因(Sul1、Sul2、Sul3)的检测[J].食品科学,2014,35(24):178-183.
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[24]董洪燕,缪晓斌,李鑫,等.鸡白痢沙门菌分离株的耐药性及磺胺类耐药机制研究[J].中国家禽,2010,32(9):29-33.
[25]廖成水,程相朝,张春杰,等.鸡源致病性沙门氏菌新近分离株的耐药性与耐药基因[J].中国兽医科学,2011,41(7):751-755.
[26]Huerta B,Marti E,Gros M,et al.Exploring the links between antibiotic occurrence,antibiotic resistance,and bacterial communities in water supply reservoirs[J].Science of the Total Environment,2013(456/457):161-170.
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[28]Ji X,Shen Q,Liu F,et al.Antibiotic resistance gene abundances associated with antibiotics and heavy metals in animal manures and agricultural soils adjacent to feedlots in Shanghai,China[J].Journal of Hazardous Materials,2012(235/236):178-185.
[29]Hoa P T P,Managaki S,Nakada N,et al.Antibiotic contamination and occurrence of antibiotic-resistant bacteria in aquatic environments of northern Vietnam[J].Science of the Total Enviromnet,2011,409(15):2894-2901.
[30]王娜.环境中磺胺类抗生素及其抗性基因的污染特征及风险研究[D].南京:南京大学,2014.
[2]Xu J,Zeng L B,Zhang H,et al.Cyprinid herpesvirus2infection emerged in cultured gibel carp,Carassius auratus gibelioin China[J].Veterinary Microbiology,2013,166(1/2):138-144.
[3]吴霆,丁正峰,朱春艳,等.异育银鲫鳃出血病流行病学调查和研究[J].水产科学,2014,33(5):283-287.
[4]叶灵通,李文祥,邹红,等.池塘养殖异育银鲫寄生黏孢子虫的种群动态[J].水生生物学报,2014,38(6):1092-1097.
[5]张晓君,阎斌伦,邴旭,等.异育银鲫病原温和气单胞菌表型及分子鉴定与溶血素基因检测[J].水生态学杂志,2010,3(4):102-107.
[6]耿昕颖,王家祯,董文龙,等.异育银鲫嗜水气单胞菌的分离鉴定与病理组织学观察[J].中国兽医杂志,2016,52(4):109-112.
[7]林溪,田国明,朱凝瑜,等.浙江地区异育银鲫内脏携带细菌多样性与毒力基因分析[J].水产科学,2014,33(12):785-789.
[8]Khushiranabi R,Girisha S K,Bhowmick P P,et al.Prevalence of different outer membrane proteins in isolates of Aeromonas species[J].Word Journal of Microbiology and Biotechnology,2008,24(10):2263-2268.
[9]Vivas J,Carracedo B,Riano,J,et al.Behavior of an Aeromonas hydrophila aroA live vaccine in water microcosms[J].Applied and Environmental Microbiology,2004,70(5):2702-2708.
[10]Fang H M,Ge H,Sin Y M.Cloning,characterization and expression of Aeromonas hydrophila major adhesion[J].Fish&Shellfish Immunology,2004,16(5):645-658.
[11]Phan T P H,Lia N,Pham H V,et al.Detection of the sul1,sul2,and sul3genes in sulfonamide-resistant bacteria from wastewater and shimp ponds of north Vietnam[J].Science of the Total Enviromnet,2008,405(1/3):377-384.
[12]吴俊伟.动物源大肠埃希菌氟喹诺酮类药物多重耐药机制与耐药水平相关性研究[D].雅安:四川农业大学,2011.
[13]彭彬,杨光友,陈晓利,等.黄鳝出血病病原菌的分离与鉴定[J].淡水渔业,2011,41(3):72-76.
[14]冷闯,邓舜洲,张文波,等.中华鳖致病性嗜水气单胞菌的分离鉴定及药敏试验[J].动物医学进展,2012,33(2):124-129.
[15]朱芝秀,何后军,邓舜洲,等.嗜水气单胞菌江西地区分离株耐药性及耐药质粒分析[J].江西农业大学学报,2012,34(6):1262-1268.
[16]黄钧,黄艳华,胡大胜,等.黄沙鳖白底板病病原菌的分离鉴定及6种毒力基因检测[J].水生生物学报,2013,37(5):844-854.
[17]刘灿.嗜麦芽窄食单胞菌对磺胺类抗菌药物耐药机制的研究进展[J].安徽医学,2014,18(5):961-963.
[18]Skold O.Sulfonamide resistance:mechanisms and trends[J].Drug Resistance Updates,2000,3(3):155-160.
[19]Enne V I,King A,Livermore D M,et al.Sulfonamide resistance in Haemophilusinfluenzae mediated by acquisition of sul2or a short insertion in chromosomal folP[J].Antimicrobial Agents and Chemotherapy,2002,46(6):1934-1939.
[20]Perreten V,Boerlin P.A new sulfonamide resistance gene(sul3)in Escherichia coli is widespread in the pig population of Switzerland[J].Antimicrobial Agents and Chemotherapy,2003,47(3):1169-1172.
[21]羊云飞,王红宁,谭雪梅,等.二重PCR检测猪、鸡源致病性大肠杆菌、沙门氏菌磺胺类耐药基因(Sul1、Sul2、Sul3)的研究[J].畜牧兽医学报,2007,38(10):1088-1092.
[22]赖海梅,刘书亮,邹立扣,等.肉鸡屠宰场多重耐药沙门氏菌Ⅰ类整合子与磺胺类耐药基因(Sul1、Sul2、Sul3)的检测[J].食品科学,2014,35(24):178-183.
[23]Antunes P,Machado J,Sousa J C,et al.Dissemination of sulfonamide resistance genes(Sul1,Sul2,Sul3)in portuguese Salmonella enterica strains and relation with integrons[J].American Society for Microbiology,2005,49(2):836-839.
[24]董洪燕,缪晓斌,李鑫,等.鸡白痢沙门菌分离株的耐药性及磺胺类耐药机制研究[J].中国家禽,2010,32(9):29-33.
[25]廖成水,程相朝,张春杰,等.鸡源致病性沙门氏菌新近分离株的耐药性与耐药基因[J].中国兽医科学,2011,41(7):751-755.
[26]Huerta B,Marti E,Gros M,et al.Exploring the links between antibiotic occurrence,antibiotic resistance,and bacterial communities in water supply reservoirs[J].Science of the Total Environment,2013(456/457):161-170.
[27]Mckinney C W,Loftin K A,Meyer M T,et al.Tet and sul antibiotic resistance genes in livestock lagoons of various operation type,configuration,and antibiotic occurrence[J].Environmental Science&Technology,2010,44(16):6102-6109.
[28]Ji X,Shen Q,Liu F,et al.Antibiotic resistance gene abundances associated with antibiotics and heavy metals in animal manures and agricultural soils adjacent to feedlots in Shanghai,China[J].Journal of Hazardous Materials,2012(235/236):178-185.
[29]Hoa P T P,Managaki S,Nakada N,et al.Antibiotic contamination and occurrence of antibiotic-resistant bacteria in aquatic environments of northern Vietnam[J].Science of the Total Enviromnet,2011,409(15):2894-2901.
[30]王娜.环境中磺胺类抗生素及其抗性基因的污染特征及风险研究[D].南京:南京大学,2014.