昆明地区鸡源沙门氏菌的分离鉴定及耐药性分析
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摘要
为了解昆明周边养鸡场的沙门氏菌耐药状况,采集昆明周边5家养殖场的50只疑似沙门氏菌病的发病鸡的肝脏和脾脏进行细菌分离,通过常规生化特性鉴定和PCR分子鉴定,然后通过药敏试验检测分离株耐药性,通过PCR技术对分离株的耐药基因进行检测。结果表明:共分离出8株沙门氏菌,8株菌的靛基质试验、尿素酶试验和VP试验结果均呈阴性,S3、S4菌株不产H2S,柠檬酸试验也呈阴性,针对沙门氏菌设计特异性引物进行扩增,得到目的条带495 bp,与预期一致;在药敏试验中,分离株对氨苄西林和头孢氨苄有较强的耐药性,对四环素中介耐药,对恩诺沙星、加替沙星、卡那霉素、庆大霉素以及氧氟沙星敏感,说明分离株已出现耐药性;在所检测的6种耐药基因中,β-内酰胺类的bla TEM-1和喹诺酮类的GyrA基因检出率为100%,四环素类的tetA基因未被检出,其他三种耐药基因aadA1、tetB和strB检出率均在50%以上,说明分离株耐药基因的携带率较高,需要引起养殖业的重视。
In order to understand the drug resistance of Salmonella in chicken farms around Kun‐ming, the liver and spleen of 50 chickens suspected of salmonellosis from 5 farms around Kunming were collected for bacterial isolation, identification of routine biochemical characteristics and molecular identification of PCR, then the drug resistance of the isolates was tested by tolerance test, the drug resistance genes of the isolates were detected by PCR. The results showed that 8 Salmonella were isolated, the results of ketole test, urease test and VP test of 8 Salmonella were negative, S3 and S4 did not produce H2 S, and citric acid test was negative. According to the specific design of the Salmonella bacteria primers were amplified to 495 bp, consistent with ex‐pectations. In the drug susceptibility test, the isolates showed strong resistance to ampicillin and cefalexin. medi-resistance to tetracycline, it is sensitive to enrofloxacin, gatifloxacin,kanamycin, gentamicin and ofloxacin. It indicated that the drug resistance of isolates had ap‐peared. Among the six drug-resistant genes detected, blaTEM-1 of beta-lactams and GyrA of quinolones were 100% and tetA of tetracyclines were not detected, the detection rates of the other three re‐sistance genes aadA1, tetB and strB were all above 50%. The results showed that the drug resis‐tance gene carrying rate of isolates was high, which should be paid attention to by the Poultry breeding industry.
In order to understand the drug resistance of Salmonella in chicken farms around Kun‐ming, the liver and spleen of 50 chickens suspected of salmonellosis from 5 farms around Kunming were collected for bacterial isolation, identification of routine biochemical characteristics and molecular identification of PCR, then the drug resistance of the isolates was tested by tolerance test, the drug resistance genes of the isolates were detected by PCR. The results showed that 8 Salmonella were isolated, the results of ketole test, urease test and VP test of 8 Salmonella were negative, S3 and S4 did not produce H2 S, and citric acid test was negative. According to the specific design of the Salmonella bacteria primers were amplified to 495 bp, consistent with ex‐pectations. In the drug susceptibility test, the isolates showed strong resistance to ampicillin and cefalexin. medi-resistance to tetracycline, it is sensitive to enrofloxacin, gatifloxacin,kanamycin, gentamicin and ofloxacin. It indicated that the drug resistance of isolates had ap‐peared. Among the six drug-resistant genes detected, blaTEM-1 of beta-lactams and GyrA of quinolones were 100% and tetA of tetracyclines were not detected, the detection rates of the other three re‐sistance genes aadA1, tetB and strB were all above 50%. The results showed that the drug resis‐tance gene carrying rate of isolates was high, which should be paid attention to by the Poultry breeding industry.
引文
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[8]李晨,张双翔,周碧君,等.腹泻仔猪肠道致病性大肠埃希氏菌和沙门氏菌的分离鉴定与耐药性研究[J].中国畜牧兽医,2017,44(12):3625-3633.
[9] Clinical and Laboratory Standards Institute.M100-S20:performance standards for antimicro‐bial susceptibility testing[S]. Wayne:CLSI,2010.
[10]邹立和,蒲妍君,杨莉,等.四川省猪肉源大肠杆菌和沙门氏菌的分离与耐药性分析[J].食品科学,2012,33(13):202-206.
[11]王晓泉,王彦红,吴双,等.四环素耐药基因在鸡源沙门氏菌中的分布和传播[J].中国家禽,2007,29(9):10-12.
[12]张正庆,罗薇,刘亚刚,等.致病性沙门氏菌对部分四环素类和氟喹诺酮类药物的耐药性研究[J].四川畜牧兽医,2007,34(1):31-32.
[2]贾爱卿.猪沙门氏菌的临床分离鉴定及耐药性消除研究[D].武汉:华中农业大学,2006.
[3]刘华伟,郭蔼光,马立农,等. PCR技术在沙门氏菌快速检测中的应用[J].动物医学进展,2004,25(6):55-58.
[4] Cohen N D, Wallis D E, Neibergs H L, et al.Comparison of the polymerase chain reaction using genus-specific oligonucleotide primers and microbiologic culture for the detection of Salmonella in drag-swabs from poultry houses[J]. Poultry Science, 1994, 73(8):1276-1281.
[5]章新生,臧富妍.应用PCR技术检测沙门氏菌[J].中国兽医学报,1999,19(2):147-151.
[6]佟金平,王镇山,薛欣,等.磷霉素与14种抗菌药物分别联用的体外抗菌活性研究[J].中国抗生素杂志,2005,30(4):233-237.
[7]侯雪娇,吴科敏,莫国东,等.食源性沙门氏菌耐药表型与耐药基因的研究[J].食品科学,2016,37(19):166-170.
[8]李晨,张双翔,周碧君,等.腹泻仔猪肠道致病性大肠埃希氏菌和沙门氏菌的分离鉴定与耐药性研究[J].中国畜牧兽医,2017,44(12):3625-3633.
[9] Clinical and Laboratory Standards Institute.M100-S20:performance standards for antimicro‐bial susceptibility testing[S]. Wayne:CLSI,2010.
[10]邹立和,蒲妍君,杨莉,等.四川省猪肉源大肠杆菌和沙门氏菌的分离与耐药性分析[J].食品科学,2012,33(13):202-206.
[11]王晓泉,王彦红,吴双,等.四环素耐药基因在鸡源沙门氏菌中的分布和传播[J].中国家禽,2007,29(9):10-12.
[12]张正庆,罗薇,刘亚刚,等.致病性沙门氏菌对部分四环素类和氟喹诺酮类药物的耐药性研究[J].四川畜牧兽医,2007,34(1):31-32.