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1.
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   

2.
红霉素与四环素耐药基因在猪链球菌临床分离株中的检测   总被引：1，自引：1，他引：0  
为了解临床分离的48株猪链球菌对大环内酯类药物及四环素耐药基因的分布,用微量稀释法测定48株临床分离的猪链球菌对大环内酯类、四环素、β-内酰胺类及头孢类9种抗生素的药物敏感性,建立PCR方法对耐药菌株大环内酯类耐药基因ermA/B/C、mefA/E、msrD、mphB、23S rRNA,L4,L22和四环素耐药基因tetM、tetO、tetL、tetK及与Tn916转座子相关的int和xis基因进行检测。结果表明,31株2型猪链球菌中大环内酯类药物耐药率为3.23%,17株9型猪链球菌红霉素耐药率为88.24%,泰乐菌素、磷酸替米考星、阿奇霉素的耐药率均为70.59%。48株猪链球菌对四环素均耐药,但对青霉素、阿莫西林、头孢曲松钠、氨苄西林均敏感。大环内酯类耐药基因主要以ermB为主,占75%(12/16),mefA/E、msrD占25%(4/16),16株红霉素耐药菌株中,tetM、tetO、int、xis的检出率分别为25%(4/16)、62.5%(10/16)、31.25%(5/16)和31.25%(5/16),没有检测到ermA、ermC、mphB、tetL、tetK。所有红霉素耐药菌株均未检测到23S rRNA、L4和L22突变。  相似文献   

3.
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   

4.
Antibiotic susceptibility of bacteria isolated from nasal swabs and lungs of pigs, to 16 commonly used antibiotics, was determined by disc diffusion test. beta-lactams showed the best activity against Streptococcus suis (S. suis) (> 99% of susceptible strains). The lowest sensitivity of S. suis was evidenced to: tylosin, tetracycline and neomycin (50%, 40% and 25%, respectively). Isolates of Escherichia coli (E. coli) demonstrated the highest susceptibility to cephalosporin (85% strains), gentamicin and norfloxacin (over 74%). The lowest susceptibility of E. coli was demonstrated to tiamulin and penicillin (11.3% and 1.9%, respectively). Over 80% of Actinobacillus pleuropneumoniae (App) strains were susceptible to all antibiotics tested. The highest resistance of App, but demonstrated by below 20% of tested isolates only, was evidenced to neomycin and LxS. Isolates of Pasteurella multocida (Pm), Haemophilus parasuis (Hps) and Arcanobacterium pyogenes (A. pyogenes) were highly susceptible to the most antibiotics included in the analysis. The comparison of the in vitro susceptibility of pathogens to the chemotherapeutics used on Polish farms for the therapy of bacterial infection of pigs within the last five years and the last 10 years, showed an increasing percent of E. coli and S. suis strains resistant to commonly used antibiotics. It is also shown that Pm, Hps, App and A. pyogenes isolates were continuously susceptible to the most chemotherapeutics applied.  相似文献   

5.
动物源性链球菌红霉素耐药基因的分布   总被引：6，自引：1，他引：6  
致病性链球菌可导致人和动物的各种化脓性疾病、肺炎、乳腺炎、败血症等，且部分致病性链球菌是重要的人畜共患病病原，对人畜健康均造成极大危害，已引起高度重视。大环内酯类抗生素作为青霉素药物的替代药物是治疗致病性链球菌感染的有效药物。国内外对人医重要的链球菌诸如肺炎球菌、化脓性链球菌的研究表明，临床分离株对大环内酯类抗生素的耐药率较高，  相似文献   

6.
化脓隐秘杆菌(Trueperella pyogenes,T.pyogenes)是一种能够引起动物和人化脓性感染的重要病原菌。随着抗生素的广泛使用,该菌已对临床常用的大环内酯类、四环素类等药物产生不同程度的耐药性。本试验拟探讨外排泵抑制剂利血平对T.pyogenes大环内酯类外排基因mefA mRNA及蛋白表达的影响,将为细菌外排泵抑制剂的研究奠定理论基础。以携带mefA基因的T.pyogenes分离株(17,20号)为试验菌株,采用荧光定量PCR及Western blot技术,检测利血平作用前后各菌株mefA基因mRNA及蛋白表达量。结果表明,1/2 MIC利血平作用2株耐药菌株36 h后,大环内酯类抗生素红霉素、罗红霉素、泰乐菌素、阿奇霉素和替米考星对携带mefA基因的T.pyogenes的MIC值均有不同程度的降低。17和20号分离株外排基因mefA mRNA表达水平较药物作用前均极显著降低(利血平作用前是作用后的5.18,17.54倍),外排蛋白MefA表达量较药物作用前也显著降低(利血平作用前是作用后的1.08,1.70倍),且基因水平和蛋白水平的变化与其耐药表型的变化趋势一致;提示利血平可能是通过抑制外排基因mefA的转录和翻译而消减T.pyogenes对大环内酯类抗生素的耐药性。  相似文献   

7.
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   

8.
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   

9.
Fifty-four quinupristin/dalfopristin-resistant Enterococcus faecium (QDREF) isolated from chickens and pigs during 2002-2003 in Korea were screened by PCR for the presence of streptogramin resistance genes vatD, vatE, and vgbA, and macrolide resistance gene ermB. None of the QDREF isolates carried vgbA and vatD genes, while vatE and ermB were detected in 9.2% and 74% of the isolates, respectively. Twenty-six percent (14/54) of the QDREF isolates contained none of the resistance determinants tested. Pulsed-field gel electrophoresis (PFGE) patterns revealed high heterogeneity: 47 different patterns for 54 QDREF evaluated. Identical PFGE types were observed in two pairs of chicken isolates and a pair of pig isolates, respectively, but chicken isolates did not share PFGE pattern with pig isolates, suggesting clonal spread of QDREF strain between the same species of animals but not between different species of animals. This is the first report, to our knowledge, of vatE-positive E. faecium isolates and also the first evidence of clonal spread of QDREF strain between animals in Korea.  相似文献   

10.
Accurate identification of mastitis pathogens is often compromised when using conventional culture-based methods. Here, we report a novel, rapid assay tested for speciation of bacterial mastitis pathogens using high-resolution melt analysis (HRMA) of 16S rDNA sequences. Real-time PCR amplification of 16S rRNA gene fragment, spanning the variable region V5 and V6 was performed with a resulting amplicon of 290bp. First, a library was generated of melt curves of 9 common pathogens that are implicated in bovine mastitis. Six of the isolates, Escherichia coli, Streptococcus agalactiae, Klebsiella pneumoniae, Streptococcus uberis, Staphylococcus aureus and Mycoplasma bovis, were type strains while the other 3, Arcanobacterium pyogenes, Corynebacterium bovis and Streptococcus dysgalactiae, were bovine mastitis field isolates. Four of the type strains, E. coli, S. agalactiae, K. pneumoniae and S. aureus, were found to be of human origin, while the other 3 type strains were isolated from bovine infections. Secondly, the melt curves and corresponding amplicon sequences of A. pyogenes, E. coli, S. agalactiae, S. dysgalactiae, K. pneumoniae, S. uberis and S. aureus were compared with 10 bovine mastitis field isolates of each pathogen. Based on the distinct differences in melt curves and sequences between human and bovine isolates of E. coli and K. pneumoniae, it was deemed necessary to select a set of bovine strains for these pathogens to be used as reference strains in the HRMA. Next, the HRMA was validated by three interpreters analyzing the differential clustering pattern of melt curves of 60 bacterial cultures obtained from mastitis milk samples. The three test interpreters were blinded to the culture and sequencing results of the isolates. Overall accuracy of the validation assay was 95% as there was difficulty in identifying the streptococci due to heterogeneity observed in the PCR amplicons of S. uberis. The present study revealed that broad-range real-time PCR with HRMA can be used as a powerful, fast and low-cost tool for the differentiation of clinically important bacterial mastitis pathogens.  相似文献   

11.
为证实国内临床分离的动物源链球菌菌株中是否存在与红霉素耐药性相关的主动外排机制，选取含有mefA基因、ermB基因、不含mefA／ermB基因的链球菌耐药菌株和青霉素、红霉素均敏感的菌株，采用完整细胞物质转运动力学方法研究了菌株对红霉素的主动外排功能。结果表明，含mefA基因的菌株对红霉素的主动外排作用显著增强，对红霉素的蓄积浓度显著低于其它菌株（P〈0．05），且氰氯苯腙（CCCP）可减少红霉素的主动外排作用，增加菌体内红霉素的蓄积浓度（P〈0．05）。本试验结果佐证了在链球菌内存在与红霉素耐药相关的主动外排机制。  相似文献   

12.
The antimicrobial susceptibility of 151 clinical Streptococcus suis strains isolated from diseased pigs in Spain was determined by a microdilution method. Isolates were mostly susceptible to beta-lactam antimicrobials, aminoglycosides, enrofloxacin, novobiocin and spectinomycin. More than 87% of the S. suis isolates were resistant to tetracyclines, sulphonamides, macrolides and clindamycin. Strains of serotype 9 were significantly more resistant than strains of serotype 2 (P<0.05) to tylosin (94% versus 77%) and clindamycin (94% versus 64%). Eighty-seven percent of the S. suis isolates were resistant to at least four antimicrobials and nine isolates (6%) were resistant to at least six antimicrobials. The most frequently identified multidrug pattern involved resistance against tetracyclines, sulphonamides, macrolides and lincosamides, with 69% of the isolates exhibiting this resistotype. Fifteen out of the 22 strains of serotype 2 (68.2%), and 84 out of the 98 of the strains of serotype 9 (85.7%) exhibited this resistotype, indicating its widespread distribution among the strains of the two most frequently isolated serotypes.  相似文献   

13.
Clostridium perfringens is a well documented cause of a mild self-limiting diarrhea and a potentially fatal acute hemorrhagic diarrheal syndrome in the dog. A recent study documented that 21% of canine C. perfringens isolates had MIC's indicative of resistance to tetracycline, an antimicrobial commonly recommended for treatment of C. perfringens-associated diarrhea. The objective of the present study was to further evaluate the antimicrobial susceptibility profiles of these isolates by determining the prevalence of specific resistance genes, their expression, and ability for transference between bacteria. One hundred and twenty-four canine C. perfringens isolates from 124 dogs were evaluated. Minimum inhibitory concentrations of tetracycline, erythromycin, tylosin, and metronidazole were determined using the CLSI Reference Agar Dilution Method. All isolates were screened for three tetracycline resistance genes: tetA(P), tetB(P) and tetM, and two macrolide resistance genes: ermB and ermQ, via PCR using primer sequences previously described. Ninety-six percent (119/124) of the isolates were positive for the tetA(P) gene, and 41% (51/124) were positive for both the tetA(P) and tetB(P) genes. No isolates were positive for the tetB(P) gene alone. Highly susceptible isolates (MIC< or = 4 microg/ml) were significantly more likely to lack the tetB(P) gene. One isolate (0.8%) was positive for the ermB gene, and one isolate was positive for the ermQ gene. The tetM gene was not found in any of the isolates tested. Two out of 15 tested isolates (13%) demonstrated transfer of tetracycline resistance via bacterial conjugation. Tetracycline should be avoided for the treatment of C. perfringens-associated diarrhea in dogs because of the relatively high prevalence of in vitro resistance, and the potential for conjugative transfer of antimicrobial resistance.  相似文献   

14.
Two type/reference strains of Brachyspira (B.) hyodysenteriae, 14 Belgian and German indole negative, and 14 Belgian, German and Swedish indole positive field isolates of strongly β-haemolytic intestinal spirochaetes were compared by pulsed-field gel electrophoresis (PFGE) patterns, biochemical reaction patterns, 16S rDNA sequences and MIC determinations of six antibacterial substances. Three tests for indole production, including a spot indole test, were compared with congruent results. All field isolates were classified as B. hyodysenteriae due to a high genetic and phenotypic similarity with the type strains. The Belgian and German indole negative isolates had identical and unique PFGE patterns for the tested restriction enzymes MluI and SalI, as well as identical 16S rDNA sequences, and they could not be differentiated by any of the methods used. Seven unique PFGE patterns were achieved from the 14 indole positive field isolates. The patterns were identical and unique for epidemiologically related isolates. Type/reference strains and isolates without known relation to other tested isolates showed unique banding patterns. The MICs of tylosin, tiamulin, erythromycin, clindamycin, carbadox and virginiamycin were determined in broth for all isolates. In contrast to Belgian and German isolates, the majority of the Swedish field isolates were susceptible to tylosin, erythromycin and clindamycin. Probable pathways of infection for some of the Swedish isolates were determined. The PFGE patterns of epidemic clones of B. hyodysenteriae remained stable for a period of up to 8 years. In vivo development of resistance to macrolide and lincosamide antibiotics due to use of tylosin was clearly indicated for two epidemic clones.  相似文献   

15.
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   

16.
Streptococcus suis infection in swine. A sixteen month study.   总被引：9，自引：0，他引：9       下载免费PDF全文
A total of 349 isolates of Streptococcus suis retrieved from different tissues from diseased pigs were examined in this study. Only 48% of them could be categorized as one of serotypes 1 to 8 and 1/2. Among typable isolates, serotype 2 was the most prevalent (23%), followed by serotype 3 (10%). The majority of all isolates originated from lungs, meninges/brain, and multiple tissues. Forty-one percent of typable isolates and 33% of untypable isolates were retrieved in pure culture. Other isolates were found in conjunction with Pasteurella multocida, Escherichia coli, Actinobacillus pleuropneumoniae, Actinomyces pyogenes, and other streptococci. Typable S. suis isolates were more frequently isolated from pigs between five and ten weeks of age, while untypable isolates were mostly found in animals aged more than 24 weeks. No obvious monthly and/or seasonal variation of the prevalence of isolation of S. suis could be detected.  相似文献   

17.
采用微量肉汤稀释法和D-试验法检测64株凝固酶阴性葡萄球菌（CNs）对10种抗菌药物的耐药性,并用PCR方法分别检测头孢西丁耐药菌株和红霉素耐药菌株中mecA基因以及erinA、ermB、ermC和msrA基因的携带情况。结果表明,所有菌株均对万古霉素和甲氧苄啶／磺胺甲恶唑敏感;泰妙菌素为耐药率（65．6％）最高的抗菌药物,其次是红霉素、氧氟沙星和β-内酰胺类药物。28株（43．8％）CNS对青霉素耐药,其中26株对头孢西丁耐药（MRS）并且均携带mecA基因。30株（46．9％）CNS对红霉素耐药,其中24株为MLSB耐药表型,主要由ermB基因介导。总之,兽医临床CNs分离株对常用抗菌药物的耐药性不同,mecA基因和ermB基因的携带分别是兽医临床MRS和MLSB表型产生的主要原因。  相似文献   

18.
2004年在病猪体内分离到一株猪链球菌,通过平板扩散法和微量稀释法药敏试验表明这株链球菌对红霉素敏感。采用这株猪链球菌进行体外诱导试验,在低浓度药物组第165代和高浓度药物组第180代的菌液对红霉素M IC值均达到中介水平。它们的耐药表型均为内在型,而且都扩增到了ermB耐药基因。其中180代菌的23S rRNA碱基1387位A突变成G;它们的核糖体蛋白L4,165代菌碱基104位、585位和633位,分别T突变成C、A突变成G和A突变成G,180代菌碱基283位和651位,分别A突变成G和T突变成G;核糖体蛋白L22,165代和180代菌碱基109位和468位,分别C突变成A和T突变成A,并且165代菌碱基还在426位G突变成A。这些碱基的突变可能是引起猪链球菌对红霉素耐药的原因之一。  相似文献   

19.
新疆北疆地区猪源粪肠球菌的耐药性分析   总被引：1，自引：1，他引：0  
为了解新疆北疆地区猪源粪肠球菌的耐药性及相关耐药基因型的分布情况,本试验采用K-B（Kirby-Baller）琼脂扩散法检测了49株猪源粪肠球菌对8种抗菌药物的敏感性,并采用PCR法对9种相关耐药基因进行检测并测序,测序结果与GenBank中的相应基因序列比对。药敏试验结果显示,分离菌对链霉素耐药率最高,其次为青霉素和红霉素,对呋喃妥因、氨苄西林高度敏感。PCR检测结果显示,β-内酰胺类耐药基因tem的检出率最高,为93.88%,其次是四环素类耐药基因tetM,为85.71%,喹诺酮类基因gyrA和parC检出率均为42.86%,氨基糖苷类耐药基因aph（3'）-Ⅲ、aac（6'）/aph2″和ant（6'）-Ⅰ的检出率分别为36.73%、16.33%和16.33%,未检出mefA和ermB基因。本试验从表型与基因型分析发现,北疆地区猪源粪肠球菌的多重耐药现象非常严重,且其耐药表型与基因型并不完全一致。  相似文献   

20.
Streptococcus agalactiae (Group B Streptococcus, GBS), Streptococcus dysgalactiae subsp. dysgalactiae (Group C Streptococcus, GCS) and Streptococcus uberis are relevant mastitis pathogens, a highly prevalent and costly disease in dairy industry due to antibiotherapy and loss in milk production. The aims of this study were the evaluation of antimicrobial drug resistance patterns, particularly important for streptococcal mastitis control and the identification of strain molecular features. Antimicrobial resistance was assessed by disk diffusion against amoxicillin–clavulanic acid, cefazolin, cefoperazone, pirlimycin-PRL, rifaximin, streptomycin, chloramphenicol, erythromycin-ERY, gentamicin, tetracycline-TET and vancomycin. Genotypic relationships were identified using pulsed-field gel electrophoresis (PFGE), macrolide and/or tetracycline resistance gene profiling, GBS capsular typing, GBS virulence gene profiling and GBS and S. uberis multi locus sequence typing (MLST).The majority of the isolates were susceptible to all drugs except to aminoglycoside, macrolide, lincosamide and tetracycline. Close to half of the TET resistant isolates have tetO and tetK and almost all ERY–PRL resistant isolates have ermB. A high degree of intra-species polymorphism was found for GCS. The GBS belonged to ST-2, -554, -61, -23 lineages and five new molecular serotypes and human GBS insertion sequences in the cpsE gene were found. Also, GBS of serotype V with scpB and lmb seem to be related with GBS isolates of human origin (same ST-2 and similar PFGE). Overall our results suggested that different therapeutic programs may have been implemented in the different farms and that in most cases clones were herd-specific.  相似文献   

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