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1.
为探究温和气单胞菌对氨基糖苷类和四环素类抗生素的耐药性,试验采用PCR法检测10株来源不同的鱼源温和气单胞菌对氨基糖苷类抗生素的4种耐药基因(aph(3')-Ⅱa、ant(3″)-Ⅰa、aac(6')-Ⅰb、aac(3)-Ⅱa)及四环素类抗生素的3种耐药基因(tetA、tetC、tetM)的表达情况,并利用K-B纸片扩散法对6种抗生素进行耐药表型分析。结果显示,10株温和气单胞菌对氨基糖苷类耐药基因aph(3')-Ⅱa、ant(3″)-Ⅰa、aac(6')-Ⅰb的检出率分别为20%、30%、20%,未检测出aac(3)-Ⅱa基因;对四环素类的耐药基因tetA、tetC、tetM的检出率分别为70%、20%、60%。K-B纸片扩散法结果显示,10株菌对四环素耐药率最高,对链霉素敏感,对庆大霉素、卡那霉素、多西环素、米诺环素高度敏感。结果表明,本次分离的温和气单胞菌对氨基糖苷类和四环素类抗生素具有一定的耐药性,为深入了解温和气单胞菌的耐药机制提供参考。  相似文献   

2.
为了解鱼源致病性小肠结肠炎耶尔森菌对氨基糖苷类药物的耐药现状,试验采用KirbyBauer纸片法检测12株鱼源小肠结肠炎耶尔森菌对3种氨基糖苷类药物的耐药表型,PCR方法分析氨基糖苷类耐药基因aph(3)-Ⅲ、ant(3″)-Ⅰ、aac(6')-Ⅰ和aac(3)-Ⅱ在致病菌中的分布情况。结果表明:12株菌株均对卡那霉素、庆大霉素高度敏感,其中1株菌株对链霉素中度敏感。4种耐药基因aph(3)-Ⅲ、ant(3″)-Ⅰ、aac(6')-Ⅰ和aac(3)-Ⅱ的检出率分别为91.7%、83.3%、83.3%、0。鱼源小肠结肠炎耶尔森菌耐氨基糖苷类药物的耐药基因检出率显著高于耐药表型,因此检测耐药基因比耐药表型更能体现该类药物的耐药现状。  相似文献   

3.
为探究温和气单胞菌对氨基糖苷类和四环素类抗生素的耐药性,试验采用PCR法检测10株来源不同的鱼源温和气单胞菌对氨基糖苷类抗生素的4种耐药基因(aph(3′)-Ⅱa、ant(3″)-Ⅰa、aac(6′)-Ⅰb、aac(3)-Ⅱa)及四环素类抗生素的3种耐药基因(tetA、tetC、tetM)的表达情况,并利用K-B纸片扩散法对6种抗生素进行耐药表型分析。结果显示,10株温和气单胞菌对氨基糖苷类耐药基因aph(3′)-Ⅱa、ant(3″)-Ⅰa、aac(6′)-Ⅰb的检出率分别为20%、30%、20%,未检测出aac(3)-Ⅱa基因;对四环素类的耐药基因tetA、tetC、tetM的检出率分别为70%、20%、60%。K-B纸片扩散法结果显示,10株菌对四环素耐药率最高,对链霉素敏感,对庆大霉素、卡那霉素、多西环素、米诺环素高度敏感。结果表明,本次分离的温和气单胞菌对氨基糖苷类和四环素类抗生素具有一定的耐药性,为深入了解温和气单胞菌的耐药机制提供参考。  相似文献   

4.
鲫鱼弗氏柠檬酸杆菌的耐药表型及耐药基因检测   总被引:1,自引:0,他引:1  
为了检测鲫鱼费氏柠檬酸杆菌的耐药表型及耐药基因,试验采用标准Kirby-Bauer纸片扩散法检测鲫鱼弗氏柠檬酸杆菌对13种抗生素的耐药表型,并用PCR法检测弗氏柠檬酸杆菌对四环素类和磺胺类抗生素的6种耐药基因(tetA、tetC、tetM、Sul1、Sul2和Sul3)。结果表明:鲫鱼弗氏柠檬酸杆菌对四环素、氨苄西林、强力霉素、红霉素、头孢拉定和阿莫西林耐药;仅检测出四环素类的tetA和tetM基因,而其他未检测出。  相似文献   

5.
旨在调查鸭致病性大肠杆菌氨基糖苷修饰酶耐药基因(AMEs基因)的携带情况,探讨耐药基因与氨基糖苷类抗生素耐药表型的相关性。对98株鸭致病性大肠杆菌采用了K-B法,选用氨基糖苷类抗生素链霉素、新霉素、庆大霉素、阿米卡星、大观霉素和卡那霉素进行药敏试验,用建立的检测氨基糖苷类AMEs主要基因的四重PCR方法对上述菌株进行分子检测,并随机选取耐药基因ant(3″)-Ⅰa、aac(3)-Ⅱa和aph(3′)-Ⅱa各3个阳性扩增进行克隆测序,对药敏试验结果和基因检测结果进行比较分析。结果表明,98株鸭致病性大肠杆菌有67株对上述氨基糖苷类药物中的一种或多种耐药,耐药率为68.4%(67/98);有49株扩增出AMEs基因,AMEs基因的检出率为50%(49/98),其中ant(3″)-Ⅰa的检出率为30.6%(30/98),aac(3)-Ⅱa为13.3%(13/98),aph(3′)-Ⅱa为3.1%(3/98),ant(3″)-Ⅰa+aac(3)-Ⅱa为2.0%(2/98)、aac(3)-Ⅱa+aph(3′)-Ⅱa为1.0%(1/98),未检出aac(6′)-Ⅰb基因;序列分析结果表明,扩增产物与GenBank中的相应序列有很高的同源性(99%);AMEs耐药基因与耐药表型的符合率为73.1%(49/67),符合率从高到低依次为大观霉素60%(3/5)、庆大霉素55%(11/20)、链霉素33.3%(22/66)、卡那霉素19%(4/21)、新霉素12.5%(1/8)、阿米卡星0%(0/3)。另外有4株细菌检测到相关耐药基因但耐药表型为敏感,而有22株耐药表型为耐药却未检测到相关耐药基因。鸭致病性大肠杆菌氨基糖苷类耐药基因以ant(3″)-Ⅰa和aac(3)-Ⅱa两种为主,耐药性与相关耐药基因的检出率基本呈正相关。  相似文献   

6.
为了分析四川地区牦牛源肺炎克雷伯氏菌耐药性并确定其耐药基因的携带情况,本试验采集了四川地区不同养殖场患呼吸道疾病牦牛肺脏、咽拭子、鼻拭子共127份病料组织,分离得到了43株肺炎克雷伯氏菌,并采用微量肉汤稀释法结合PCR扩增法检测43株肺炎克雷伯氏菌的耐药性和耐药基因携带情况。结果表明:43株肺炎克雷伯氏菌对氨苄西林、阿莫西林、多西环素、磺胺间甲氧嘧啶4种药物耐药率较高,为65.12%~90.70%;对阿米卡星、氟苯尼考、多黏菌素B、大观霉素4种药物的耐药率在20.93%~48.84%之间;对头孢噻呋、恩诺沙星、环丙沙星3药物的耐药率在18.60%~25.58%之间;多数筛选分离得到的菌株均呈现出多重耐药性,耐9、10种药物的菌株最多,分别占分离菌株的23.3%和20.9%。通过使用PCR扩增法检测分离菌携带的耐药基因情况,结果表明:blaTEM、blaSHV、sul2、sul3、floR 5种耐药基因检出率较高,检出率在62.8%~69.8%之间;ant (3″)-Ⅰa、aph (3')-Ⅱa、aac (6')-Ⅰb、aacC2 4种耐药基因检出率较低,检出率在7.0%~20.9%;blaCTX-M、Mcr-1、qnrA、qnrS、rmtB 5种耐药基因未检出。综上所述,本试验中分离得到肺炎克雷伯氏菌具有较强的耐药性并携带有较多的耐药基因,为了更好的促进四川省牦牛养殖产业的发展,应注意避免滥用抗生素并通过药敏试验对症下药。  相似文献   

7.
《畜牧与兽医》2017,(5):134-141
为了解禽致病性大肠杆菌耐药表型及耐药基因的情况,选取江苏、安徽等地分离的53株禽致病性大肠杆菌,采用药敏纸片法对9种抗菌药物进行药敏试验,并对四环素类tet(A)、tet(B)、tet(C)、tet(W)、tet(M)、tet(O)、tet(K)、tet(L)耐药基因,喹诺酮类GryA、ParC耐药基因,磺胺类sulⅠ、sulⅡ、sulⅢ耐药基因,β-内酰胺类SHV、CTX-M、ompCA耐药基因,氨基糖苷类aac(3)-Ⅰ、aac(3)-Ⅱ、aac(6')-Ⅰb、aac(6')-Ⅱ、ant(3″)-Ⅰ、armA、rmtA、rmtB、rmtC、rmtD、npmA耐药基因进行PCR检测。结果显示:53株禽致病性大肠杆菌对磺胺异噁唑、环丙沙星、氨苄西林、多西环素的耐药率较高,分别为88.68%(47/53)、71.71%(38/53)、86.79%(46/53)、75.47%(40/53)。其中50株禽致病性大肠杆菌表现为多重耐药,耐4、5、6种药物的现象最为普遍,且不同地区菌株存在差异。tet(A)是四环素耐药基因中最为流行的一种耐药基因(52.83%,28/53),喹诺酮类耐药基因主要由gryA(94.33%,50/53)、parC(94.33%,50/53)基因编码,耐磺胺类药物sulⅠ、sulⅡ、sulⅢ基因均有检出,分别为96.23%(52/53)、98.11%(48/53)、86.79%(46/53),耐β-内酰胺类药物中仅检出ompC A基因(30.19%,16/53),在检测的11种耐氨基糖苷类耐药基因中,最为流行为aac(3)-Ⅱ、aac(6')-Ⅰb、ant(3″)-Ⅰ基因,分别为92.59%(49/53)、98.11%(52/53)、100%(53/53)。耐药基因与相关耐药菌株检出率基本呈正相关。试验结果表明:53株禽致病性大肠杆菌耐药性高,耐药谱广,耐药基因流行现象十分普遍。本试验结果能为禽致病性大肠杆菌的耐药现状与临床用药提供理论指导。  相似文献   

8.
研究大雁、斑头雁和天鹅源沙门菌的耐药性及其耐药基因的携带情况。采用K-B纸片法对33株大雁、斑头雁和天鹅源沙门菌进行20种抗菌药物敏感性测定,应用PCR对12种耐药基因的携带情况进行检测。结果表明,33株沙门菌分离株对阿莫西林、萘啶酸耐药性最高,分别为93.94%、87.88%,对氨苄西林(54.55%)、四环素(48.49%)、多西环素(48.49%)、卡那霉素(60.61%)、庆大霉素(48.49%)、链霉素(66.67%)耐药性比较严重,对氧氟沙星、培氟沙星、诺氟沙星敏感。共检测到9种耐药基因,其中blaCMY-2阳性率为87.88%,qnr、aac(6′)-Ib-cr阳性率分别为17.24%、86.21%,tetA、tetB阳性率分别为45%、20%,aph(3)Ⅱ、aac(3)Ⅱ、aadA1阳性率分别为100%、10%、20%,sul2阳性率为100%,未检出blaTEM-1、tetC、tetG基因。说明大雁、斑头雁和天鹅源沙门菌严重耐药,且携带多种耐药基因。  相似文献   

9.
动物源粪肠球菌对7种抗生素耐药表型及耐药基因检测   总被引:4,自引:0,他引:4  
为查明动物源(牛、羊、猪、鸡)粪肠球菌分离株对7种常见抗生素的耐药情况(包括耐药表型及相关的耐药基因),采用药敏纸片法、浓度稀释法和VITEK-AMS全自动药敏法3种不同的方法,根据CLSI(2007)判定标准,检测40株分离株的耐药表型;采用聚合酶链反应(PCR)方法,检测分离株中β-内酰胺类抗生素耐药相关基因(TEM)、氨基糖苷类抗生素耐药相关基因(aac(6’)/aph2’’,aph(3’)-Ⅲ,ant(6)-I)、四环素耐药相关基因(tetM)、红霉素耐药相关基因(ermB,mefA)和万古霉素耐药相关基因(vanA,vanB,vanC)。结果表明,分离株对庆大霉素、链霉素、四环素、红霉素、青霉素、阿莫西林表型耐药率分别为:60.0%(24/40),57.5%(23/40),52.5%(21/40),67.5%(27/40),60.0%(24/40),55.0%(22/40),本试验中未发现耐万古霉素粪肠球菌。耐药基因aac(6’)/aph2’’,ant(6)-Ⅰ,aph(3’)-Ⅲ,tetM,ermB,TEM的检出率分别为:55%(22/40),55%(22/40),25.0%(10/40),42.5%(17/40),50.0%(20/40),45.0%(18/40);未检测到mefA,vanA,vanB,vanC基因的菌株。动物源性粪肠球菌多重耐药现象严重,携带抗生素相关耐药基因是导致分离株对抗生素产生耐药的主要原因,从耐药表型和基因型的角度均可证实分离株粪肠球菌具有多重耐药性。  相似文献   

10.
猪肠道菌氨基糖苷类药物耐药基因分析   总被引:4,自引:0,他引:4  
为调查和分析猪肠道菌的氨基糖苷类药物耐药机制,用PCR及序列分析方法检测鉴定2个猪肠道菌中4种16S rRNA甲基化酶耐药基因rmtA、rmtB、rmtC和armA和10种氨基糖苷钝化酶基因,用接合试验研究rmtB基因和10种氨基糖苷钝化酶基因的水平转移机制,用微量稀释法测定携带rmtB基因的分离菌及其接合子对20种抗菌药物的敏感性.结果,从分离的152株猪肠道菌中共检测到49株(32.3%)rmtB阳性菌,其中48株为大肠埃希氏菌,1株为阴沟肠杆菌,未检出其它16S rRNA甲基化酶编码基因.49株rmtB阳性菌中检测到7种氨基糖苷类钝化酶基因,检出率从高到低依次是aac(3)-Ⅱ(87.8%)、aph(3′)-Ⅶ(79.6%)、aph(3′)一Ⅱ(77.6%)、aadAJ(34.7%)、aacc(6′).-Ⅰb(14.3%)、aac(3)-Ⅳ(10.2%)和aph(4)-Ⅰ(8.2%).46株rmtB阳性菌可以通过接合试验将rmtB、aac(3)-Ⅱ、aph(3′)-Ⅶ、aph(3′)-Ⅱ和aadA基因传递给受体菌E.coli C600和E coli 488 Rifr,接合率从3.0×10-6~4.6×10-13不等.所有rmtB阳性菌株及其接合子对卡那霉素、阿米卡星、妥布霉素、西梭米星、萘替米星、庆大霉素6种氨基糖苷类抗生素均高度耐药(MIC≥512 Pg·mL-1).研究结果表明,rmtB基因和氨基糖苷钝化酶基因广泛存在于两猪场,它们共同介导猪源肠道菌对庆大霉素、阿米卡星等氨基糖苷类药物的高度耐药.rmtB基因与aac(3)-Ⅱ·aph(3′)一Ⅶ、aph(3′)-Ⅱ和aadA基因位于同一接合性质粒上,共同传播氨基糖苷类的耐药性.  相似文献   

11.
To detect the drug resistance genotype and phenotype produced by Aeromonas veronii isolated from different fish.PCR method was used to detect the aminoglycoside antibiotics resistance genes (aac (3)-Ⅱa, aac (6')-Ⅰb, ant (3")-Ⅰa, aph (3')-Ⅱa), sulfonamides resistance genes (Sul1, Sul2, Sul3) and tetracyclines resistance genes (tetA, tetC, tetM) in isolates. The drug susceptibility to seven Aeromonas veronii strains toward 22 kinds of antibiotics were performed with Kirby-Bauer disc diffusion method. The results showed that the detected resistance genes were aac (3)-Ⅱa (71.4%), aac (6')-Ⅰb (85.7%), Sul2 (85.7%) and tetA (28.5%),while the other resistant genes of ant (3")-Ⅰa, aph (3')-Ⅱa, Sul1, Sul3, tetC and tetM were not checked out. The seven Aeromonas veronii isolates were sensitive to Fosfomycin (100%), Polymyxin B (100%), Furazolidone (85.7%) and Ofloxacin (71.4%). They were resistant to Ampicillin (100%), Acetylspiramycin (100%), Trimethoprim/Sulfamethoxazole (85.7%), Sulfisoxazole (85.7%) and Tetracycline (85.7%). Drug resistance genotype had some correlation with the drug resistance phenotype.  相似文献   

12.
为指导临床合理用药并为治疗患病水貂提供依据,本试验对山东省某水貂养殖场送检的7只疑似患有沙门氏菌病的病死水貂的肝脏和脾脏样品进行细菌分离鉴定及药物敏感性分析,通过分离纯化方法从样品中分离菌株,并采用革兰氏染色、生化鉴定和PCR方法对分离菌株进行鉴定。运用K-B药敏法检测菌株对临床常用药物的敏感性,并通过PCR方法检测菌株耐药基因及Ⅰ类整合子的携带情况。结果显示,本试验分离得到的7株菌均为革兰氏阴性、短小的杆菌;生化反应检测显示,分离菌株葡萄糖、麦芽糖、甘露醇、MR试验、枸橼酸盐、硫化氢试验均表现为阳性,初步鉴定分离菌株为沙门氏菌;PCR产物测序结果进一步表明7株分离菌均为沙门氏菌;药敏试验结果显示,7株沙门氏菌对头孢曲松、左氧氟沙星、氟苯尼考和多黏菌素较敏感,对氨基糖苷类药物、四环素和氨苄西林表现为耐药;耐药基因检测结果显示,7株沙门氏菌共检测出8种耐药基因blaTEM-1、blaCTX-M-1G、aadA1、aac(3')-Ⅳ、aac(3')-Ⅱc、aph(4')-Ⅰa、aph(3')-Ⅶ和oqxAB,以及基因盒为aadA1、arr-3-aacA4和blaPSE-1的Ⅰ类整合子。综上可知,本试验在送检的7只患病水貂的肝脏和脾脏样品中分离到7株沙门氏菌,分离菌株均为耐药菌株,且主要表现为多重耐药现象;耐药基因呈多样性,且存在位于质粒上的耐药基因扩大了耐药基因的传播范围,增加了细菌的耐药性,为临床用药治疗带来困难。  相似文献   

13.
In order to determine the drug resistance phenotype and prevalence of drug resistance genes in Escherichia coli (E.coli) isolated from sheep in Inner Mongolia, the minimum inhibitory concentrations (MICs) of the isolates to antibiotics commonly used in veterinary clinical were detected by micro-dilution method in vitro. The results showed that the highest resistance rates of the isolates to amoxicillin,cephalothin,sulfamethoxazole and polymyxin were up to 100.0%,respectively.To amoxicillin-clavulanic, tetracycline and ciprofloxacin were 50% to 80%. These isolates were sensitive to cefotaxime, meropenem trihydrate, neomycin, and their resistance rates were all less than 10%. Among the 108 strains of E.coli from sheep, 94.4% of them were resistant to more than 7 antimicrobial agents,15.6% of them were resistant to 13 antimicrobial agents, only one strain was sensitive to all antimicrobial agents. Six kinds of resistance genes among the 108 E.coli isolates were detected by PCR method.The results showed that detection rates of 4 kinds of drug resistance genes including blaTEM, proP-2, sul-Ⅰ and ampG were all over 50%,the detection rate of resistance gene aph (3')-Ⅰ was up to 40%, only resistance gene aac(3)-Ⅱ detection rate was 5.5%. Thus, the sheep E.coli isolates in Inner Mongolia produced various degrees of resistance to 13 kinds of antibiotics, and their multi-drug resistances were very serious. They carried aph(3')-Ⅰ, sul-Ⅰ, ampG, blaTEM, proP-2 and aac(3)-Ⅱ resistance genes.  相似文献   

14.
张楠驰  方庆  王利 《中国畜牧兽医》2019,46(6):1832-1840
试验旨在确定感染黄颡鱼的病原菌并探讨其耐药性。采用细菌培养、生理生化鉴定及16S rDNA序列分析方法进行菌株分离鉴定,运用PCR方法扩增耐药基因,用纸片扩散法对分离菌进行药敏试验。结果显示,分离菌与门多萨假单胞菌基本一致,16S rDNA基因长度为1 442 bp。同源性及系统进化树分析显示,该菌16S rDNA序列与门多萨假单胞菌NK-01同源性为99.6%,亲缘关系最近。综合判定分离菌为门多萨假单胞菌,命名为fsznc-01。耐药基因检测结果显示,分离菌fsznc-01含有β-内酰胺类耐药基因TEM,氨基糖苷类耐药基因aph(3')-Ⅱa、aac(6)-Ⅰb、aac(3)-Ⅱa及磺胺类耐药基因Sul1和Sul2,未检测出磺胺类耐药基因Sul3。药敏试验结果显示,分离菌fsznc-01对头孢他啶、庆大霉素和哌拉西林等药物敏感,对头孢氨苄、头孢唑林和头孢拉定耐药。本试验结果可为预防门多萨假单胞菌感染导致的鱼类疾病提供参考依据。  相似文献   

15.
In order to provide therapeutical guidance for drug admistration, the bacteria of three sick minks suffering from typical diarrhea symptoms provided by mink farms in Jilin province were isolated and identified, and the drug sensitivity was tested. The bacteria were isolated with TSA plates, and identified using biochemical methods and PCR assay. The virulence of the isolates was determined by infecting BALB/c mice. The antimicrobial susceptibility of the isolates to antimicrobial agents was investigated using the K-B method. PCR was used to detect the resistance genes and Ⅰ integrons. A total of 3 Shigella isolates were obtained from sick minks. The virulent determination showed that all isolates could cause mice diarrhea. The drug sensitivity results showed that 3 strains were sensitive to fluoroquinolone, cephalosporin, florfenicol and polymyxin, but they were resistant to aminoglycoside, tetracycline, chloramphenicol, penicillin and ampicillin. There were seven resistance genes were detected,blaTEM-1,aadA1, aac(3')-Ⅱc, aac(6')-Ⅰb, aph(3')-Ⅶ, tet(M), cat2 and three class Ⅰ integrons carrying aadA 1 gene cassette. All of the isolates were virulent and caused the mice diarrhea. The resistance of the 3 strains were very serious and mainly for multiple drug resistance phenomenon. The resistance genes detected in the mink were various, and could bring enormous implications for clinical treatment.  相似文献   

16.
【目的】研究广东省茂名地区屠宰环节猪肉中携带的沙门氏菌的耐药性和毒力特征,为该地区食源性沙门氏菌的危害评估和防控措施制定提供依据。【方法】从广东省茂名地区屠宰场采集的猪肉、脾脏、肝脏样本中分离到19株沙门氏菌,采用K-B药敏纸片法检测其对β-内酰胺类、氟喹诺酮类、氨基糖苷类、四环素类、酰胺醇类和磺胺类抗菌药物的耐药性,用PCR方法检测β-内酰胺类耐药基因(blaTEM、blaOXA-1、blaSHV)、氟喹诺酮类耐药基因(qnrA、qnrS、qnrB)、氨基糖苷类耐药基因(aadA1、aac(6')-Ⅰb、rmtB)、四环素类耐药基因(tetA、tetB、tetC)、酰胺醇类耐药基因(Cat1、floR)、磺胺类耐药基因(SulⅠ、SulⅡ、SulⅢ)和10种毒力基因(mogA、sseL、mgtC、bcfA、araB、spvR、spvA、spvB、spvC、spvD)的携带情况。【结果】19株沙门氏菌耐药严重,对四环素、多西环素、氯霉素、氟苯尼考、磺胺异噁唑的耐药率均>50%,对四环素、多西环素和氯霉素的耐药率最高,均为89.5%(17/19),对3种及其以上抗菌药物耐药率为89.5%,最高对11种抗菌药物均有耐药性;blaTEMqnrAaadA1、aac(6')-ⅠbCat1、floRSulⅠ、SulⅢ耐药基因的检出率较高(≥50.0%),blaTEM耐药基因检出率最高为84.2%,同时携带5个及以上耐药基因的菌株占73.7%,最高携带12个耐药基因;mogAmgtCbcfAaraB毒力基因均有检出,且检出率均在70%以上,其中mogA基因的检出率高达100%,其余6种毒力基因均未检出。【结论】广东省茂名地区屠宰场猪肉中沙门氏菌多重耐药严重,携带多种耐药基因且具有复杂的基因组合类型,同时携带多种毒力基因。  相似文献   

17.
为了解鸡源致病性大肠埃希菌对氨基糖苷类抗生素的耐药性变化和钝化酶耐药基因的携带情况及耐药基因与耐药性的相关性,从陕西、河南、河北、山西、宁夏和甘肃6省(区)的部分规模化养鸡场的病、死鸡中分离鉴定320株致病性大肠埃希菌。采用K-B药敏纸片法检测分离菌对6种氨基糖苷类药物的敏感性,PCR方法检测6种氨基糖苷类钝化酶耐药基因,用DNA Star软件对获得的耐药基因序列与GenBank中的相关序列进行比对。结果显示,鸡源致病性大肠埃希菌分离株对庆大霉素、链霉素、妥布霉素、卡那霉素、新霉素和阿米卡星的耐药率分别为53.4%、49.3%、37.5%、34.7%、22.8%和5.3%,对妥布霉素和卡那霉素耐药率呈上升趋势,而对庆大霉素的耐药率虽呈下降趋势,但仍维持在40%以上。3重以上耐药菌株占80%(256/320)。氨基糖苷类钝化酶基因aac(3)-Ⅱ、aph(3′)-Ⅰ和aac(6′)-Ⅰ的检出率分别为50.9%、25.9%和3.1%,未检测到aac(3)-Ⅳ、ant(3′′)-Ⅰ和aph(3′)-Ⅱ基因。研究表明,分离的鸡源致病性大肠埃希菌对氨基糖苷类抗生素的耐药性普遍存在,以多重耐药为主,且对妥布霉素和卡那霉素的耐药性不断上升。耐药基因aac(3)-Ⅱ和aph(3′)-Ⅰ的检出率与其耐药性呈正相关。  相似文献   

18.
To further identify the pathogenic strains and analyze their antibiotic resistance, the methods of pathogen isolating, the methods of Gram staining, biochemical tests and 16S rRNA PCR amplification, virulence tests, drug sensitive tests, virulence genes and resistance genes PCR amplification were used. The results showed that the strain was Gram-positive and revealed positive after reacting with 6.5% NaCl, melibiose, sucrose and etc., while revealed negative reactions with VP, hippurate, arabinose and etc.. It showed 100% similarity with Enterococcus feacium 16S rRNA gene sequence in GenBank after PCR amplification of 16S rRNA gene and BLAST alignment. It was found severe pathogenicity after virulence tests in mice. The strain was highly resistant to oxacillin, penicillin G of β-lactam antibiotics and norfloxacin, ciprofloxacin, levofloxacin of quinolones antibiotics and tetracycline, while was sensitive to antibiotics of erythromycin, vancomycin and clindamycin. It revealed that virulence factor genes Asal, cylA, acm and resistance genes TetM, ant(6)-Ⅰ, aac(6')-aph (2"), ermB showed positive. The results showed that the bacteria was Enterococcus feacium, it had a strong pathogenicity and severe drug resistance, which might be related to the highly positive rates of virulence genes and drug resistance genes.  相似文献   

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