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1.
为分析奶牛乳房炎中分离的金黄色葡萄球菌(S.aureus)四环素耐药机制,本研究以K-B法和常量肉汤法测定最低抑菌浓度(MIC)值,并对26株奶牛乳房炎S.aureus进行四环素耐药表型检测及加入利血平后的MIC值检测;以PCR检测四环素耐药基因tetM、tetO、tetL和tetK,对扩增产物进行序列分析。检测结果表明:26株菌株中,7株对四环素耐药,19株敏感;利血平能显著降低部分耐药株对四环素的MIC值;在7株耐药菌株中,1株检测出tetM基因,6株检测出tetK基因,没有检测到tetL和tetO基因;tetK基因与S.aureus质粒pT181同源性为100%,tetM基因与转座子Tn916序列同源性为99.9%。实验研究表明,26株奶牛乳房炎S.aureus四环素耐药表型与耐药基因相符,耐药机制以外排蛋白介导为主,并在国内牛源S.aureus中首次检测到tetK基因。 相似文献
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为了解猪源链球菌对红霉素相关耐药基因ermB、ermA和mefA的分布,对河北省及辽宁部分地区的64株猪源链球菌分离株,用PCR方法检测51株红霉素耐药株和13株红霉素敏感株中ermB、ermA和mefA基因。结果显示,耐药菌株中ermB基因的检出率为98.04%(50/51),ermA的检出率为25.49%(13/51),没有检出mefA基因。初步表明河北省及辽宁部分地区的猪源链球菌对红霉素的耐药机制以ermB基因介导为主。20株菌的ermB基因核苷酸序列与GenBank中同源序列相似性为99%～100%。与参照序列AJ972604.1相比,20株菌的ermB氨基酸序列的突变位点较少,主要有Thr 75→Ser、Ser 100→Asn、Arg 118→His、Leu 175→Ile,以100和118位突变为主,进一步说明ermB基因是相对稳定的。 相似文献
3.
东北部分地区猪链球菌对四环素类药物耐药机制的研究 总被引：5，自引：0，他引：5
为了研究东北部分地区猪链球菌对四环素类药物的耐药机制,在兰氏分群的基础上,对东北部分地区分离的12株猪链球菌进行四环素类药物的耐药性测定和相关基因的扩增。结果表明:12株猪链球菌均属于D群,其中91.7%对多西环素耐药,100%对土霉素耐药,83.3%对盐酸金霉素耐药;12株菌均扩增出tetM基因,但未扩增出tetO基因。同源性分析结果显示扩增的tetM基因与GenBank中的肺炎链球菌、粪肠球菌等的tetM基因的同源性为94%~100%。 相似文献
4.
《中国兽医学报》2015,(7):1130-1135
对江苏地区多个奶牛场采样,并对奶牛乳房炎(Bovine mastitis,BMT)检测阳性奶样进行病原菌分离、鉴定。对分离出的凝固酶阴性葡萄球菌(Coagulase-negative Staphylococci,CNS)分别进行20种药物的敏感性试验和相关耐药基因检测。结果显示,BMT检测阳性的奶样有610份,528份检出细菌,共检出细菌802株,鉴定为CNS的217株;大部分CNS对万古霉素、左氟沙星、氯霉素、磷霉素、呋喃妥因敏感,对青霉素、大观霉素具有较强耐药性;CNS对β-内酰胺类(blaZ)、大环内酯类(ermC)和四环素类(tetK、tetM、tetL、tetO)耐药基因的检出率分别为29.95%、2.30%、24.88%、1.84%、1.84%和0.00%。 相似文献
5.
猪链球菌对红霉素耐药性的研究 总被引：2，自引：0，他引：2
从发病猪体内分离、鉴定猪链球菌，采用肉汤稀释法和纸片琼脂扩散法筛选对红霉素耐药的猪链球菌，用双纸片法确定耐药株的耐药表型，通过聚合酶链反应检测对红霉素耐药的基因ermb/mefA。猪链球菌对红霉素的耐药表型为cMLS表型，即同时对克林霉素耐药。在3株红霉素耐药株中扩增到ermb基因，其余未能检测到ermb或mefA基因。 相似文献
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7.
为了确定保定市某猪场发病猪感染的病原并了解该病原的生物学特性，试验对采集的肺脏组织病料进行病原菌的分离，采用荧光定量PCR、生化试验对分离菌株进行鉴定，并对分离菌株进行血清型鉴定、药敏试验、耐药基因和毒力基因检测。结果表明：分离到一株病原菌，将其命名为BDS21。经鉴定该分离菌株为21型猪链球菌。BDS21菌株对头孢曲松钠、阿奇霉素敏感，对氨苄西林、卡那霉素、左氧氟沙星中介，对头孢噻肟、四环素、林可霉素、环丙沙星耐药；检出tetM和ermA 2种耐药基因，gapdh、sly、fbps、gdh、orf2 5种毒力基因。说明该养殖场发病猪感染的病原可能是21型猪链球菌，防控首选药物为头孢类和大环内酯类抗生素。 相似文献
8.
为了解内蒙古地区奶牛乳房炎链球菌毒力基因的分布情况及其耐药现状,通过该地区360份乳房炎奶样链球菌的分离鉴定,利用微量肉汤稀释法测定了81株链球菌对15种药物的MIC,采用PCR法检测耐药基因及毒力基因的携带情况。结果显示,分离到的81株链球菌对β-内酰胺类的耐药率最高;对红霉素和四环素的耐药率在40%~85%;对氟喹诺酮类的耐药率相对较低。多重耐药菌株达88.9%(n=72)。耐药基因pbp2b的检出率为86.4%,tetL和tetM的检出率在75%以上,红霉素耐药表型主要由ermB介导。分离菌株毒力基因中cfb的检出率最高(50.6%),其次为cyl和hylB。内蒙古地区奶牛乳房炎链球菌对β-内酰胺类、大环内酯类和四环素耐药情况严重,且发现多重耐药菌株的流行;链球菌中尤以无乳链球菌的毒力基因携带率较高。 相似文献
9.
广东地区猪链球菌的耐药性特点及四环素耐药基因携带情况 总被引：2，自引：2，他引：0
猪链球菌(Streptococcus suis,SS)是危害世界养猪业的重要细菌性病原菌之一,本研究旨在了解猪链球菌临床分离株的耐药性特点和四环素耐药基因的携带情况,为临床上该病的防控提供科学依据。本试验采用K-B法和CLSI推荐的肉汤微量稀释法检测猪链球菌广东分离株对18种抗菌药物的耐药性。结果显示,被检菌株对四环素类、磺胺类和氨基糖苷类抗菌药物表现较强的耐药性,尤其对四环素类药物的耐药率高达98.2%;对头孢菌素类药物比较敏感;菌株都耐受6种或6种以上的抗菌药物,其中以6和14重耐药菌株的数量最多,分别占20%和17%。本试验设计了4对引物检测四环素耐药基因携带情况,结果发现56株菌中以携带tetM基因为最多(85.71%)。结果表明tetM很可能是介导广东SS分离株对四环素产生极高耐药率的主要原因之一。 相似文献
10.
猪链球菌对大环内酯类药物耐药性研究进展 总被引：1，自引：0，他引：1
随着养猪业的迅速发展,猪链球菌病也在世界范围内广泛传播。大环内酯类药物在养猪业中的广泛应用也使其耐药性随之产生,国内外猪链球菌对大环内酯类的耐药性较严重,耐药率达50%以上。目前国内外已报道的猪链球菌对大环内酯类耐药基因包括ermA、ermB、ermC和mefA,其中最常见的是ermB基因编码的内在型（cMLSB）耐药。大环内酯类耐药性特点表现为多重耐药株（MDR）占很大比例且与菌株血清型之间可能存在相关性。耐药基因erm及mef均为获得性耐药基因,位于转座子内,或由质粒携带,可在细菌间广泛传播。合理使用抗菌药物,加强耐药性监测,开发新药以及疫苗等对于减缓耐药性的产生和传播具有十分重要的意义。 相似文献
11.
Antibiotic resistance in animal isolates of enterococci is of public health concern because of the risk of transfer of antibiotic resistance isolates or resistance determinants to consumers via the food chain. In this study, phenotypic and genotypic resistance in 192 pig isolates of enterococci to ampicillin, avilamycin, avoparcin, bacitracin, flavophospholipol, gentamicin, narasin, tetracycline, tiamulin, tylosin, vancomycin, virginiamycin, copper and zinc were investigated by susceptibility test and molecular methods. Resistance rates varied between the species but all isolates were susceptible to ampicillin, avilamycin, avoparcin, gentamicin and narasin but resistant to tetracycline and tylosin and intermediately resistant to copper. Only Enterococcus gallinarum and Enterococcus casseliflavus were resistant to vancomycin and virginiamycin resistance was present in less than half the Enterococcus faecium isolates. Zinc resistance was largely confined to Enterococcus faecalis but bacitracin resistance was uncommon in E. faecalis in comparison with the other species. Tiamulin resistance was common in all species except E. casseliflavus. Resistance to flavophospholipol was detected in most E. faecium isolates and in a high proportion of E. gallinarum, E. casseliflavus and E. hirae/durans but was only found in one isolate of E. faecalis. No tetO, rplC, rplD, vanA, vanB, vatA and vatD genes were found. The presence of ermB, tetL, tetM, tcrB, aac6-aph2, tetK, tetS, vanC1, vanC2, lsaA, lsaB and vatE varied between the species and largely corresponded to the susceptibility phenotype. The findings show that resistance to antibiotics of high clinical significance for nosocomial Enterococcus infections is absent, whereas antimicrobial resistance was detected for some other antibiotics including bacitracin, flavophospholipol, tetracycline, tiamulin, tylosin and virginiamycin. 相似文献
12.
Prevalence and mechanism of resistance against macrolides and lincosamides in Streptococcus suis isolates 总被引：13，自引：0，他引：13
Martel A Baele M Devriese LA Goossens H Wisselink HJ Decostere A Haesebrouck F 《Veterinary microbiology》2001,83(3):287-297
Eighty-seven Streptococcus suis isolates recovered in 1999-2000 from diseased pigs, all from different farms, were screened for resistance against macrolide and lincosamide antibiotics by the disk diffusion and agar dilution test and a PCR assay, amplifying the ermB gene and the mefA/E gene. Seventy-one percent of the isolates showed constitutive resistance to macrolide and lincosamide antibiotics (MLS(B)-phenotype). All these isolates were positive for the ermB gene in the PCR, but negative for the mefA/E gene. For all strains minimum inhibitory concentrations (MIC) against five other antimicrobial agents were determined. All strains were susceptible to penicillin. Ninety-nine percent of the isolates were susceptible to enrofloxacin and tiamulin. Eighty-five percent of the strains were resistant to doxycycline. A 540bp fragment of the ermB genes of eight S. suis strains was sequenced and compared with ermB genes of five S. pneumoniae and five S. pyogenes strains of human origin. A 100% homology was found between these fragments in seven S. suis, one S. pneumoniae and three of the S. pyogenes isolates. This study demonstrates that resistance against macrolides, lincosamides and streptogramin B is widespread in S. suis and mediated by ribosome methylation, encoded by the ermB gene. 相似文献
13.
Prevalence of mecA, blaZ, tetO/K/M, ermA/B/C, aph, and vanA/B/C/D genes conferring resistance to oxacillin, penicillin, tetracycline, erythromycin, gentamicin, and vancomycin was investigated in 65 staphylococcal isolates belonging to twelve species obtained from ready-to-eat porcine, bovine, and chicken products. All coagulase negative staphylococci (CNS) and S. aureus isolates harbored at least one antibiotic resistance gene. None of the S. aureus possessed more than three genes, while 25% of the CNS isolates harbored at least four genes encoding resistance to clinically used antibiotics. In 15 CNS isolates the mecA gene was detected, while all S. aureus isolates were mecA-negative. We demonstrate that in ready-to-eat food the frequency of CNS harboring multiple antibiotic resistance genes is higher than that of multiple resistant S. aureus, meaning that food can be considered a reservoir of bacteria containing genes potentially contributing to the evolution of antibiotic resistance in staphylococci. 相似文献
14.
Environmental streptococcus isolates from bovine milk were identified to the species and strain level and screened for resistance to macrolide and lincosamide antibiotics by phenotypic and genotypic methods. Isolates were tested for resistance to erythromycin and pirlimycin by broth microdilution assays. Presence of ribosomal methylase genes (ermA, ermB, ermC) and efflux pump genes (mefA/E, msrA/C) was detected by polymerase chain reaction (PCR). Resistance to pirlimycin (minimum inhibitory concentration (MIC) = 8microg/ml) was detected in 6 of 13 Enterococcus isolates that were identified as E. faecium by API20Strep typing. msrC was detected in 10 enterococcal isolates but the detection of msrC was not associated with phenotypic resistance. msrC negative isolates were reclassified as Enterococcus mundtii based on sequencing of housekeeping genes. Resistance to erythromycin and pirlimycin (MIC > 16microg/ml) was detected in 4 of 4 Streptococcus dysgalactiae and 12 of 20 Streptococcus uberis isolates and was encoded by ermB. All Streptococcus isolates tested negative for ermA, ermC, mefA/E and msrA/C. Among ermB positive streptococci, three alleles were identified based on a 527 bp gene fragment. Each allele was detected in at least two herds. The same alleles have also been detected in other bacterial species from bovine and non-bovine hosts and farm soil, suggesting a theoretical potential for horizontal transfer of macrolide resistance genes on dairy farms. 相似文献
15.
Aarestrup FM AgersŁ Y Ahrens P JŁrgensen JC Madsen M Jensen LB 《Veterinary microbiology》2000,74(4):353-364
The species distribution, susceptibility to 19 antimicrobial agents and presence of selected genes encoding resistance to macrolides, streptogramins and tetracyclines were examined among 118 staphylococcal isolates from infections of poultry in Denmark. Isolates were identified using a combination of conventional biochemical testing and 16S rDNA sequencing. The most common species were Staphylococcus aureus (83), Staphylococcus hyicus (11), Staphylococcus xylosus (9) and Staphylococcus cohnii (6). The isolates were susceptible to most antimicrobials tested. A high frequency of S. aureus (30%) was resistant to ciprofloxacin. Only six (7%) S. aureus isolates and one Staphylococcus saprophyticus were penicillin resistant. Resistance to sulphamethoxazole was observed among 16 (19%) of S. aureus isolates and two coagulase negative staphylococci (CNS). Twenty (24%) of the S. aureus isolates were resistant to erythromycin and 19 of these isolates contained the ermA gene, whereas the remaining isolate contained the ermC gene. Eleven (48%) of the novobiocin resistant CNS were resistant to erythromycin and all these isolates contained the ermA gene. Two isolates identified as S. xylosus, were found to be resistant to streptogramins and both contained the vatB- and the vgaB-genes. Thirty-nine (47%) of the S. aureus isolates, three of nine S. hyicus and eight of the 23 novobiocin resistant CNS were tetracycline resistant and all contained the tet(K) gene. A single S. aureus isolate also contained the tet(M) gene. The present study showed a frequent occurrence of resistance to fluoroquinolones, tetracycline and macrolides among staphylococci isolated from broilers in Denmark, whereas the occurrence of resistance to other antimicrobial agents remains low. Similar genes, encoding resistance to erythromycin, tetracycline and streptogramins to those previously observed, were detected. 相似文献
16.
为了解甘肃某地区奶牛运动场环境中主要耐药性细菌的分布情况及耐药基因的携带情况,试验采用抗性平板筛选法分离、纯化耐药菌后再通过16S rRNA鉴定;采用PCR方法检测6类22种耐药基因携带情况。结果显示,两年两个奶牛运动场（A和B）分离出不同种属的耐药菌共665株,经16S rRNA PCR鉴定主要为大肠杆菌、屎肠球菌和普通变形杆菌,其中耐药性大肠杆菌数量最多（398株）,A牛场耐药菌数两年相比增长28.50%,B牛场耐药菌数量持平;通过对样品进行10类药物筛选耐药菌结果显示,耐药率前3位的分别为糖肽类（11.13%,74/665）、四环素类和磺胺类（10.98%,73/665）及β-内酰胺类和氨基糖苷类（10.23%,68/665）。采用PCR方法对样品进行耐药基因检测发现,两年两个奶牛运动场的耐药基因携带情况基本相同,喹诺酮类耐药基因qnrB、qnrS的携带率分别为10.81%和83.78%,磺胺类耐药基因Sul1、Sul2和Sul3的携带率分别为100%、100%和56.76%,糖肽类耐药基因VanA、VanB和VanC的携带率分别为78.38%、100%和100%,四环素类耐药基因tetA、tetB、tetC、tetM、tetO、tetL的携带率分别为100%、100%、100%、100%、81.08%和75.68%,KPC、NDM、aac（6'）-Ⅰb-cr耐药基因的携带率分别为32.43%、29.73%及24.32%,未检出qnrC、tetK、tetW、VIM和OXA耐药基因。携带耐药基因与GenBank中登录参考株基因序列的同源性为98%~99%。甘肃地区奶牛运动场中耐药菌对常用抗菌药物普遍耐药且携带相应耐药基因,奶牛运动场中碳青霉烯类及氨基糖苷类耐药基因与上年结果相比数量增加,各相关单位人员应对其加强奶牛场中耐药菌的监测,临床治疗中应合理用药,以降低环境中耐药菌转移给人类的潜在风险。 相似文献
17.
采用微量稀释法测定36株2型猪链球菌对四环素的耐药性,应用PCR扩增四环素相关耐药基因tet（M）、tet（O）、tet（K）、tet（L）、tet（Q）、tet（S）、tet（T）和tet（W）,将扩增到的耐药基因克隆、测序,并进行序列分析。结果显示,36株2型猪链球菌对四环素的耐药率为100%,MIC90高于512mg/L;其中29株扩增出tet（M）基因,6株扩增出tet（O）基因,5株同时扩增到tet（M）、tet（L）基因,同时扩增到tet（M）、tet（L）基因的菌株MIC均高于512mg/L;同源性分析结果显示,扩增到的tet（M）基因与GenBank中已公布序列的同源性为95%～100%,tet（O）基因与GenBank中已公布序列的同源性为95%～99%。结果表明,我国大部分地区的2型猪链球菌对四环素均具有很强的耐药性,主要耐药机制是由tet（M）基因介导的核糖体保护作用。 相似文献