养殖场空气中E. coli磺胺类抗生素的抗性
Characterization of sulfa antibiotics resistance of E. coli from the air of poultry farms
-
摘要: 本文从养殖场空气中分离出360株E. coli(大肠杆菌,Escherichia coli),应用肉汤微量稀释法和PCR方法,分离磺胺甲唑敏感菌株,检测抗生素抗性和抗性基因.在分离的E. coli中,对磺胺甲唑敏感菌株为95株(26.4%),有48株含有青霉素、氯霉素、四环素、环丙沙星、庆大霉素和利福平的抗性,而47株未含有抗性.其中,7株菌株含有1种抗生素抗性、11株菌株含有2种抗生素抗性、17株菌株含有3种抗生素抗性、6株菌株含有4种抗生素抗性、4株菌株含有5种抗生素抗性、3株菌株含有6种抗生素抗性.对抗生素的耐受能力依次为:氯霉素、青霉素、四环素、环丙沙星、庆大霉素、利福平.磺胺甲唑敏感菌株共检出163个抗性基因,sul1、int1、sul2、Int2、sul3检出数量依次为49、44、29、20和19;含一种、二种、三种、四种、五种抗性基因菌株分别为45、22、10、7、2;但有6株未检测出抗性基因.结果表明养殖场建场时间、抗生素使用、养殖规模等与抗生素抗性菌的抗性呈正相关;养殖场空气中分离的E. coli抗生素抗性较高,且具有多重抗性;抗生素抗性的表现型与其基因型之间出现不完全吻合现象.Abstract: 360 Escherichia coli(E. coli)samples were isolated from the air of eight poultry farms. Microdilution and polymerase chain reaction(PCR) were used to evaluate the sulfonamides susceptibility and detect the sulphonamides resistance gene respectively. The results showed that 95 samples were found to be susceptible to sulfamethoxazole, and of which 48 samples showed resistance to at least one of six different kinds of antibiotics. Briefly, 7 samples were shown to be resistant to one antibiotics,11 strains were resistant to two kinds of antibiotics, 17sampleswere resistant to three kinds of antibiotics, 6sampleswere resistant to four kinds of antibiotics, 4sampleswere resistant to five kinds of antibiotics, and notably 3 strains were resistant to all six kinds of antibiotics. The resistance abilities of the samples from highest to lowest were against to chloramphenicol, penicillin, tetracycline, ciprofloxacin, and rifampicin. Moreover, 163 resistance genes were detected from isolated sulfonamide resistance samples. There were 49 resistance genes of sul1, 44 resistance genesof sul2, 20 resistance genes of sul3, 20 resistance genes of int1 and 19 resistance genes of int2.In addition, there were 45 stains contained one kind of resistance genes, 22 contained two kinds of resistance genes, 10 contained three kinds of resistance genes,7 contained four kinds of resistance genes, and 2 contained five kinds of resistance genes. But 6 samples had no resistance genes mentioned above. Our study showed that the antibiotic resistance of bacteria was positively correlated with many factors, such as built time, antibiotic use, and breeding scale etc. The E. coli isolated from the air in the poultry farm was more resistant to antibiotics, and displayed multiple antibiotic resistances. Obviously, the sulfonamides resistance did not correlate with the presence of such genes.
-
Key words:
- sulfanilamide resistance bacteria /
- ARGs /
- E. coli /
- air in the poultry farm.
-
[1] 高敏, 贾瑞志, 仇天雷,等. 集约化养鸡场空气环境中生物气溶胶特点研究[J]. 农业环境科学学报, 2015, 34(4):787-794. GAO M, JIA R Z,QIU T L, et al. Characteristics of bioaerosols in air environment of confined poultry feeding operations[J]. Journal of Agro-Environment Science, 2015, 34(4):787-794(in Chinese).
[2] GREGOVA G, KMETOVA M, KMET V, et al. Antibiotic resistance of Escherichia coli isolated from a poultry slaughterhouses[J].Ann Agric Environ Med, 2012, 19(1):75-77. [3] 贺小萌, 曹罡, 邵明非, 等. 空气中抗性基因(ARGs)的研究方法及研究进展[J]. 环境化学, 2014, 33(5):739-747. HE X M, CAO G, SHAO M F, et al. Research method and progress on antibiotics resistance genes (ARGs) in air[J].Environmental Chemistry, 2014, 33(5):739-747(in Chinese).
[4] CHEN B W, LIANG X M, HUANG X P, et al. Differentiating anthropogenic impacts on ARG s in the pearl river estuary by using suitable gene indicators[J]. Water Research, 2013, 47:2811-2820. [5] 徐冰洁, 罗义, 周启星,等. 抗生素抗性基因在环境中的来源、传播扩散及生态风险[J]. 环境化学, 2010, 29(2):169-178. XU B J, LUO Y, ZHOU Q X, et al. Sources, dissemination, and ecological risks of antibiotic resistances genes (ARGs) in the environment[J].Environmental Chemistry, 2010, 29(2):169-178(in Chinese).
[6] 房文艳, 高新磊, 李继, 等. 城市社区农贸市场空气微生物及抗生素抗性基因研究[J]. 生态毒理学报,2015, 10(5):95-99. FANG W Y, GAO X L, LI J, et al. Investigation on airborne bacteria and antibiotic resistance genes in urban community farmers market[J]. Asian Journal of Ecotoxicology, 2015, 10(5):95-99(in Chinese).
[7] JUST N A, LÉTOURNEAU V, KIRYCHUK S P, et al. Potentially pathogenic bacteria and antimicrobial resistance in bioaerosols from cage-housed and floor-housed poultry operations[J]. Annals of Occupational Hygiene, 2012, 56(4):440-449. [8] HONG P Y, LI X, YANG X, et al. Monitoring airborne biotic contaminants in the indoor environment of pig and poultry confinement buildings[J]. Environmental Microbiology, 2012, 14(6):1420-1431. [9] 柳敦江. 养鸡场舍环境携带耐药基因的金黄色葡萄球菌的气溶胶形成及传播研究[D].济南:山东农业大学, 2012. LIU D J. Chicken farm environment carry resistant gene of staphylococcus aureus formation and propagation of aerosol research[D]. Jinan:Shandong Agricultural University, 2012(in Chinese). [10] LING A L, PACE N R, HERMANDEZ M T, et al. Tetracycline resistance and class 1 integron genes associated with indoor and outdoor aerosols[J]. Environmental Science & Technology, 2013, 47(9):4046-4052. [11] 高新磊, 邵明非, 贺小萌, 等. 污水处理厂空气介质抗生素抗性基因的分布[J]. 生态毒理学报,2015, 10(5):89-94 GAO X L, SHAO M F, HE X M, et al. Distribution of airborne antibiotic resistance gene in wastewater treatment plant[J]. Asian Journal of Ecotoxicology, 2015, 10(5):89-94(in Chinese).
[12] Clinical and laboratory standards institute (CLSI). Method for broth dilution susceptibility testing of bacteria isolated[S]. Wayne, PA, USA:CLSI.2006. [13] National committee for clinical laboratory standards. NCCLS Document M1002S12, 2002b. Performance standards for antimicrobial susceptibility testing:Twelfth informational supplement[S]. [14] 冀秀玲, 刘芳, 沈群辉, 等. 养殖场废水中磺胺类和四环素抗生素及其抗性基因的定量检测[J]. 生态环境学报, 2011, 20(5):927-933. JI X L, LIU F, SHEN Q H, et al. Quantitative detection of sulfonamides and tetracycline antibiotics and resistance genes in sewage farms[J]. Ecology and Environmental Sciences, 2011, 20(5):927-933(in Chinese).
[15] 金明兰,刘凯,徐莹莹,等.污水处理厂中磺胺类抗生素抗性菌、抗性基因的特性[J]. 环境工程, 2015, 33(11):1-5. JIN ML, LIU K, XU YY, et al. Characteristics of sulfonamide antibiotics, resistant bacteria, resistance genes in sewage treatment plant[J]. Environmental Engineering, 2015, 33(11):1-5(in Chinese).
[16] MARSHALL B M, LEVYS B. Food animals and antimicrobials:Impacts on human health[J].Clinical Microbiology Reviews, 2015, 24(4):718-733. [17] 隋倩雯,张俊亚,魏源送,等.畜禽养殖废水生物处理与农田利用过程抗生素抗性基因的转归特征研究进展[J].环境科学学报, 2016, 36(1):16-26. SUI Q W, ZHANG J Y, WEI Y S, et al. Fate of antibiotic resistance genes in the process of biological treatment and land application of animals wastewater:An overview[J].Acta scientiae Circumstantiae, 2016, 36(1):16-26(in Chinese).
[18] HVISTENDAHL M. China takes aim at rampant antibiotic resistance[J].Science,2012, 336(6083):795. [19] 张晓丹,高丽丽,胡家卿, 等.猪源携带质粒介导喹诺酮类耐药基因大肠杆菌气源性传播的研究[J].中国畜牧兽医, 2015, 33(7):739-747. ZHANG X D, GAO L L, HU J Q,et al. Porcine carrying plasmid mediated class quinolone resistant gene of E. coli air sexually transmitted[J]. Journal of China ANIMAL Husbandry and Veterinary, 2015, 33(7):739-747(in Chinese).
[20] FANG H, HAN Y, YIN Y, et al. Variations in dissipation rate, microbial function and antibiotic resistance due to repeated introductions of manure containing sulfadiazine and chlortetracycline to soil.[J].Chemosphere, 2014, 96:51-56. [21] RUTGERSSON C, GUNNARSSON L, FICK J, et al. Oral exposure to industrial effluent with exceptionally high levels of drugs does not indicate acute toxic effects in rats[J]. Environ Toxicol Chem, 2013, 32(3):577-584. [22] 李楠. 养殖场空气中细菌分布及耐药性研究[D].北京:中国人们解放军军事医学科学院, 2013. LI N. Farms in the air bacteria distribution and drug resistance study[D].Beijing:The Chinese People'S Liberation Army Military Academy of Medical Sciences, 2013(in Chinese). [23] SU HC, YING GG, TAO R, et al. Class 1 and 2 integrons, sul resistance genes and antibiotic resistance in Escherichia coli isolated from Dongjiang River, South China[J]. Environmental Pollution, 2012, 169:42-49. [24] FINLEY RL, CONLLIGON P, LARSSON DG J, et al. The scourge of antibiotic resistance:The important role of the environment[J]. Clin Infect Dis, 2013, 57(5):704-710. [25] LOPES GV, PISSETTI C, DACRUZ P P D, et al. Resistance phenotypes and genotypes of Salmonella enterica subsp. enterica isolates from feed, pigs, and carcasses in brazil[J].J Food Prot, 2015, 78(2):407-413. [26] BRAYKOV NP, EISENBERG JN, GROSSMAN M,et al. Antibiotic resistance in animal and environmental samples associated with small-scale poultry farming in northwestern ecuador.[J]. Msphere, 2016,10(1):1-15. [27] SIDRACH-C R, BÉCARES E. Fecal indicator bacteria resistance to antibiotics in experimental constructed wetlands[J]. Ecological Engineering, 2013, 50:107-111. [28] GAO P, MAO D Q, LUO Y, et al. Occurrence of sulfonamide and tetracycline resistant bacteria and resistance genes in aquaculture environment[J]. Water Research, 2012, 46:2355-2364. [29] 王娜,杨晓洪,郭欣妍, 等. 磺胺类耐药菌中抗性基因sul的表达规律[J]. 生态毒理学报,2015, 10(5):75-81. WANG N, YANG X H, GUO X Y, et al. Expression patterns of sul genes in sulfonamide-resistant bacteria[J]. Asian Journal of Ecotoxicology, 2015, 10(5):75-81(in Chinese).
[30] 秦丽婷,童蕾,刘慧,等.环境中磺胺类抗生素的生物降解及其抗性基因污染现状[J]. 环境化学,2016,35(5):875-883. QIN LT, TONG L,LIU H,et al.Biodegradation of sulfonamides and the pollution characteristics of sulfonamide resistance genes in the environment[J].Environmental Chemistry,2016,35(5):875-883(in Chinese).
[31] JI X L, SHEN Q H, 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, 235:178-185.
计量
- 文章访问数: 896
- HTML全文浏览数: 826
- PDF下载数: 469
- 施引文献: 0