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为了更好地了解禽源沙门菌对四环素的耐药性及耐药基因分布,从不同来源的家禽样品中分离沙门菌,调查其对四环素的耐药性以及耐药菌株中8种四环素耐药基因[tet(A)、tet(B)、tet(C)、tet(W)、tet(M)、tet(D)、tet(K)和tet(L)]的携带情况.结果表明,18.8%的沙门菌分离株对四环素耐药,健康成鸡分离株对四环素的耐药性明显高于雏鸡、死胚或病禽分离株;四环素耐药株中tet(A)、tet(B)和tet(M)基因的携带比例分别为73.1%、11.5%和3.8%,说明沙门菌对四环素的耐药机制以tet(A)和tet(B)基因介导的主动外排为主.本研究首次在对四环素耐药的沙门菌中检测到tet(M)基因,说明tet(M)基因在沙门菌对四环素的耐药性方面也具有潜在的作用. 相似文献
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东北地区猪链球菌对大环内酯类药物耐药机制的研究 总被引：1，自引：0，他引：1
采用微量稀释法及双纸片扩散法测定了28株猪链球菌（S．suis）对大环内酯类药物的耐药性和耐药表型，红霉素、阿奇霉素和泰乐菌素耐药率分别为92．8％、92.8％和89.3％，耐药表型以内在型（cMLSB）为主。利用聚合酶链反应（PCR）检测红霉素耐药基因erm和mef,对ermA／B／C分型扩增、克隆、测序，26株菌扩增到ermB基因，16株菌扩增到ermA基因，其中15株同时扩增到ermB和ermA基因，1株未扩增到这两种基因的任一种；28株猪链球菌中均未扩增到ermC和mef基因。16株菌的c17nA基因核苷酸序列与GenBank中同源序列同源性为83％～100％，氨基酸序列与参照序列（X03216，1）相比突变点较多，有11株菌在34个位点处同时发生了改变；26株菌的ermB基因核苷酸序列与GenBank中同源序列相似性为98％～100％，氨基酸序列与参照序列（AY183117．1）相比突变点较少，与参照序列完全一致的有7株，其余株仅1～3个氨基酸不同。说明本地区猪链球菌对大环内酯类药物耐药情况很严重，耐药基因为甲基化酶ermA和／或ermB，ermA基因突变点较多，ermB基因相对稳定。 相似文献
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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
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《畜牧与兽医》2015,(12):94-97
采用琼脂二倍稀释法测定256株水禽大肠杆菌分离株对四环素类药物的敏感性,并通过PCR方法调查分离株携带耐药基因tet A、tet B、tet C和tet M的情况。药敏试验结果表明256株水禽大肠杆菌分离株对四环素和多西环素耐药率分别为92.2%和77.3%。在237株大肠杆菌四环素耐药株中,tet A、tet B、tet C和tet M的携带率分别为49.8%、57.8%、49.4%和40.1%,仅8.0%的耐药株没有检测到4种耐药基因。结果表明:tet A、tet B、tet C和tet M广泛存在于水禽源大肠杆菌中。tet A和tet B对水禽源大肠杆菌四环素耐药株的产生起重要作用,主动外排作用是大肠杆菌对四环素产生耐药性的主要机制。 相似文献
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通过微量稀释法药敏试验及四环素类耐药基因的扩增对18株不同源多杀性巴氏杆菌进行耐药性分析。并采用PCR及其产物测序检测上述菌株中一类整合酶携带率。结果表明,18株不同源的多杀性巴氏杆菌对氯霉素的耐药率为83.33%,对四环素、强力霉素与庆大霉素的耐药率为50%~61.11%,对氧氟沙星、卡那霉素、恩诺沙星与氟苯尼考的耐药率为27.78%~38.89%。18株菌株均未检测到tet A耐药基因,22.22%的多杀性巴氏杆菌携带tet B耐药基因,5.56%的多杀性巴氏杆菌携带tet O耐药基因,tet K与tet Q两种耐药基因的携带率均为27.28%,而对于tet G的携带率为100%,Ⅰ型整合酶携带率为22.22%。本研究为进一步研究多杀性巴氏杆菌耐药机制及跨物种传播奠定了基础。 相似文献
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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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【目的】了解广东地区猪链球菌（Streptococcus suis，S.suis）临床分离株对四环素类抗生素的耐药性及耐药基因携带情况。【方法】采用药敏纸片扩散法对2016－2020年分离自广东地区的34株猪链球菌进行四环素类抗生素耐药分析，并通过PCR方法检测四环素耐药基因tetA、tetC、tetD、tetM、tetO和tetX携带情况。【结果】34株猪链球菌对四环素的耐药率高达100%（34/34），其次为米诺环素82.4%（28/34）、多西环素47.1%（16/34）。34株猪链球菌中3重耐药菌株有15株，占比44.1%（15/34），2重耐药菌株有14株，占比41.1%（14/34），耐1种药物的菌株5株，占比14.7%（5/34）。耐药基因检测结果显示，tetA、tetC、tetD、tetM、tetO和tetX基因携带率分别为0、0、0、14.7%（5/34）、100%（34/34）和0。【结论】广东地区猪链球菌临床分离株四环素类抗生素的耐药率较高，主要携带的四环素耐药基因为tetO和tetM基因。 相似文献
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猪源致病性沙门氏菌四环素耐药基因tetC的PCR扩增及序列分析 总被引：2，自引：0，他引：2
四环素类抗生素作为一类广谱抗生素,现已产生严重的耐药性.作为人兽共患病原菌的沙门氏菌,也存在大量这样的耐药菌株,使得本病的发病率和死亡率都不断上升.本研究对规模化猪场分离的经动物试验和生化试验鉴定的致病性沙门氏菌16株、药敏质控菌ATCC25922和沙门氏菌标准株C79-13对进行了四环素、金霉素、土霉素的药敏试验,结果表明16株致病性沙门氏菌对四环素类抗生素表现出普遍耐受性,耐药率达100%.其中MIC值＞128μg/mL的高耐药菌株13株,高耐药率81.25%;MIC值为32μg/mL的低耐药菌株3株,低耐药率18.75%.设计沙门氏菌四环素抗性基因tetC的引物,对16株致病性沙门氏菌的tetC基因做PCR扩增,结果12株菌获得以质粒为模板长约400bp的特异性扩增产物,未能从染色体扩增到产物.分别对临床分离的1株低耐药菌株(DY1)和1株高耐药菌株(SL1-3)的tetC基因扩增产物进行序列分析,结果表明菌株DY1和SL1-3的tetC的核苷酸同源率为97.7%;菌株DY1与质粒PBR322中tetC的核苷酸同源率为98.7%;菌株SL1-3与质粒pBR322中tetC的核苷酸同源率为98.4%,说明对于耐药程度和地方来源各不相同的菌株,在核苷酸序列上同源率很高.本文用PCR技术对规模化猪场分离的致病性沙门氏菌的四环素耐药基因进行了研究,以期对四环素耐药性的分子流行病学进行监测,克服了药敏试验只能检测耐药表型的缺点,为有效控制沙门氏菌感染提供理论基础和科学依据,在兽医、食品卫生和公共卫生等方面具有重要的意义. 相似文献
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Nasal swabs were collected at three time points from 2378 calves in four feedlots and cultured for Histophilus somni to assess genetic relatedness and tetracycline resistance. The proportions of animals carrying tetracycline resistant isolates were 0.32% at arrival, 14.82% at interim, and 0.80% at exit. The 606 H. somni isolates recovered were compared by pulsed-field gel electrophoresis (PFGE), screened for the presence of plasmids, and assessed for the tetracycline resistance genes tet(A), tet(B), tet(C), tet(E), tet(G), tet(H), tet(K), tet(L), tet(M) and tet(O) using multiplex polymerase chain reaction. Most of the isolates (98.6%) belonged to one of seven PFGE clusters (A-G) of closely related profiles with 77.7% of the isolates belonging to clusters C and D. Clusters A, B and E were associated with a higher proportion of tetracycline susceptible isolates. Genetic diversity of the isolates was highest at entry in the feedlot and lowest after the period when the animals received in-feed chlortetracycline (interim samples). Clusters A and E were more prominently represented at exit from the feedlot than other clusters. All resistant strains harboured the gene tet(H) while no other tetracycline resistance genes and no plasmids were detected with the methodology employed. It appears that genetic variability in H. somni in Alberta feedlots is low, dissemination likely occurs by clonal expansion, and resistance to tetracyclines is mediated by the tet(H) encoded efflux pump. Pulsotypes associated with tetracycline susceptible strains appear more common at exit suggesting that the in-feed oxytetracycline included throughout the feeding period is not sufficient to exert selective pressure for resistant strains. 相似文献
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Kim TJ Na YR Lee JI 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2005,52(3):119-124
A total of 160 Staphylococcus intermedius isolates were recovered from cases of pyoderma in 2002 and were examined for susceptibility to 13 different antimicrobial agents. Ninety per cent (144) of the isolates were resistant to tetracycline, derivatives of which have been used until recently, and 18% (29) were resistant to chloramphenicol which was banned from use 13 years ago. The presence of genes encoding chloramphenicol acetyltransferase (CAT) and tetracycline resistance (tet); tet(K), (L), (M), and (O) were determined by PCR in the 29 chloramphenicol and tetracycline resistant isolates. Seventeen (59%) isolates contained the cat gene while 12 (41%) isolates did not carry the cat gene, implying there may be other genes for chloramphenicol resistance that were not detected by the primers (primer set 1) used in this study. The tet(M) gene was found in 28 (97%) of the resistant S. intermedius isolates, but none contained the tet(O) gene. All 29 isolates carried one or two tet genes; tet(K), (L), and (M), with four different distribution patterns. New PCR products, a 1.1 kb product using primer set 1 and a 0.2 kb product using primer set 2, were cloned and sequenced. A 904 bp fragment of S. aureus plamid pS194, including sequence from the streptomycin adenyltransferase gene (804 bp), was found inserted into the terminal region of the cat gene (GenBank accession no. AY604739), whilst the sequence of 0.2 kb was previously unpublished. 相似文献
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Paenibacillus larvae is the causal agent of American Foulbrood (AFB) disease, the most virulent bacterial disease of honeybee (Apis mellifera L.) brood. Oxytetracycline is the main antibiotic used for prevention and control of AFB. Using the polymerase chain reaction, isolates were screened for the presence of the tetracycline resistance tet(K) and tet(L) determinants. Four isolates (5%), which correlated with the Tc-resistant phenotypes, were found to carry the tet(K) determinant, whereas none carried the tet(L) determinant. P. larvae cells were also screened for the presence of extrachromosomal DNA and evidence obtained that tetracycline resistance is plasmid-encoded. A few P. larvae isolates were found to be able to transfer the tet(K) determinant to Bacillus subtilis, suggesting that a conjugation mechanism may be involved in the transfer of the tetracycline-resistant phenotype. Minimum inhibitory concentrations to tetracycline were determined for 75 isolates of P. larvae from different geographical origins and found to range between 0.062 and 128 microg tetracyclineml(-1), with MIC(50) and MIC(90) values of 1 and 4, respectively. According to results from P. larvae populations, isolates could be considered as susceptible when their MICs were <4, intermediate for MICs values 4-8 and resistant for MICs > or = 16. To our knowledge, this is the first report of Tc(r)Paenibacillus species carrying a tet(K) gene, and also the first record of P. larvae strains carrying tet(K) determinants and its correlation with the presence of extrachromosomal DNA. 相似文献
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From 1996 to 2001 a total of 467 Staphylococcus hyicus isolates from exudative epidermitis (EE) in pigs in Denmark were examined for susceptibility to 13 different antimicrobial agents. The presence of selected genes encoding macrolide (erm(A), erm(B) and erm(C)), penicillin (blaZ), streptogramin (vat, vga, vga(B), vat(B), vat(D) and vat(E)), streptomycin (aadE) and tetracycline resistance (tet(K), tet(L), tet(M) and tet(O)) were determined in selected isolates.The occurrence of erythromycin resistance increased from 33% in 1996 to a maximum of 62% in 1997 and decreased to 26% in 2001. Resistance to sulphametazole increased from 17% in 1996 to 30% in 1998 but has since decreased to 4% in 2001. Resistance to trimethoprim increased to 51% in 1997 and decreased to 21% in 2001. Resistance to tetracycline (21-31%) remained relatively constant during 1996-2000, but increased to 47% in 2001. Resistance to penicillin (54-75%) streptomycin (33-53%) and tetracycline (21-47%) remained relatively constant over the time investigated.All 48 penicillin resistant isolates examined contained the blaZ gene and 40 (85%) of the streptomycin resistant isolates the aadE gene. It was not possible to detect any streptogramin resistance gene in four streptogramin resistant isolates. Of the 55 erythromycin resistant isolates examined, five contained erm(A), 13 erm(B), 35 erm(C) and two both erm(A) and erm(C). The presence of erm(B) was confirmed by hybridization to plasmid profiles in all 13 PCR-positive isolates. Of 52 tetracycline resistant isolates examined, two contained tet(L), 38 tet(K) and 12 both tet(K) and tet(L). 相似文献
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The aim of the study was to investigate antimicrobial susceptibilities of bovine Helcococcus ovis isolates and to detect genes encoding for H. ovis erythromycin and tetracycline resistance. Twenty-nine isolates were included and the minimal inhibitory concentrations (MICs) of seven antimicrobials were determined using test conditions as given in an approved CLSI guideline for the pyridoxal-dependent Abiotrophia spp. and Granulicatella spp. Furthermore, the macrolide resistance phenotype was examined by the erythromycin-clindamycin double-disk test (D-zone test). Erythromycin MICs of ≥ 8 μg/ml were found in three (10%) isolates which also presented the macrolide, lincosamide, and streptogramin B (MLS(B)) resistance phenotype, either constitutive or inducible. The erm(B) gene was detected in one of these isolates. Increased tetracycline MICs (≥ 8 μg/ml) were obtained for 24 (83%) isolates, mostly associated with the tet(M) gene alone (n=21) or both the tet(L) and tet(M) genes (n=2). The MICs determined for penicillin, ampicillin, amoxicillin-clavulanic acid, and cephalothin did not indicate resistance to these antimicrobials. The study suggests that resistance to MLS(B) antimicrobials and tetracycline is frequent in H. ovis. Moreover, this is the first report about occurrence of the resistance genes erm(B), tet(L), and tet(M) in the Helcococcus genus. 相似文献
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Isolates of Streptococcus suis serotype 7 from diseased pigs in Denmark were characterized by ribotyping, pulsed field gel electrophoresis (PFGE), MIC-determinations and detection of resistance genes. Forty-one different ribotype profiles were found among the 103 isolates and could be divided into two main clusters. No obvious relationship between ribotypes and the clinical origin of the isolates could be observed. Fifty-four isolates, including all 24 isolates belonging to the main ribotype profile were examined by PFGE and 50 different profiles were found. A high frequency of resistance to erythromycin (41%), tetracycline (24%) and streptomycin (28%) was observed. Furthermore, almost all isolates (101) were resistant to sulphamethoxazol. Most isolates were susceptible to ceftiofur, chloramphenicol, florfenicol, penicillin, ciprofloxacin, trimethoprim and trimethoprim + sulphonamides. The tet(M) gene was found among 11 (44%) and the tet(O) gene in six (24%) of 25 tetracycline resistant isolates. The tet(L) and tet(S) genes were not detected in any isolates. The erm(B) gene was detected in 39 (93%) of 42 erythromycin resistant isolates. 相似文献
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Vancraeynest D Hermans K Martel A Vaneechoutte M Devriese LA Haesebrouck F 《Veterinary microbiology》2004,101(4):245-251
Fifty-six Staphylococcus aureus isolates recovered between 1998 and 2003 from 31 rabbit farms with and without problems of chronic staphylococcosis, were screened for resistance to enrofloxacin, erythromycin, gentamicin, lincomycin, neomycin, penicillin and tetracyclines using the agar dilution test. For penicillin, a disk diffusion test was also performed. The detection of tetP(B), tet(K), tet(L), tet(M), tet(O), tet(T), tet(W), erm(A), erm(B), erm(C) and mec(A) genes was done via a PCR assay. Four isolates showed resistance to erythromycin and lincomycin. These isolates were positive for the erm(C) gene in the PCR. Eleven strains were resistant to tetracyclines and all harboured the tet(K) gene. In the agar dilution test, five isolates showed resistance to penicillin, whereas in the disk diffusion test 12 isolates showed resistance. None of these 12 resistant isolates carried the mec(A) gene. Only one strain showed resistance to gentamicin, and all strains were susceptible to enrofloxacin and neomycin. This study demonstrates that resistance to antimicrobial agents in S. aureus isolates originating from rabbits is relatively rare compared to resistance in S. aureus isolates originating from other animals and humans. 相似文献
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Yamamoto K Sasaki Y Ogikubo Y Noguchi N Sasatsu M Takahashi T 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2001,48(4):293-301
This is the first report to demonstrate the presence of tet(M) in naturally occurring isolates of tetracycline-resistant Erysipelothrix rbusiopathiae, which causes swine erysipelas. The tet(M) gene was isolated from E. rhusiopathiae strain KY5-42. The nucleotide and the deduced amino acid sequence were 99% identical to the tet(M) gene from Enterococcus faecalis. The gene was necessary and sufficient for the expression of tetracycline resistance in Escherichia coli. The presence of the tet(M) gene in the 114 tetracycline-resistant E. rhusiopathiae isolates from diseased pigs was detected by the polymerase chain reaction assay. The specific amplified DNA fragment was obtained from all 114 tetracycline-resistant strains. It was suggested that the tet(M) gene was widely present in the field isolates of E. rhusiopathiae resistant to tetracycline. 相似文献
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Tetracycline (tet) resistance in Campylobacter isolated from organically raised broilers was investigated in this study. Two hundred forty-five samples from an organic broiler farm were collected weekly from the first week to the end of the production cycle, and they were cultured for thermophilic Campylobacter. Tetracycline resistance of these Campylobacter isolates was identified by the agar dilution method, whereas DNA fingerprinting profiles of tet-susceptible and tet-resistant strains were determined by pulsed-field gel electrophoresis (PFGE). None of the Campylobacter isolates from the third and the fourth week of the production period were resistant to tetracycline, whereas 66.7% of the isolates from the fifth week were resistant to this antibiotic. Although the prevalence of tetracycline resistance reached 100.0% during the sixth and seventh week, less than 34.0% of the isolates from the 10th week were resistant to this antimicrobial agent. In addition, only 13.8% of Campylobacter isolates from the intestinal tracts of these organically raised broilers were resistant to tetracycline. The presence of the tet(O) gene was detected in 98.9% of tet-resistant Campylobacter isolates, and tet-susceptible and tet-resistant Campylobacter strains showed distinct PFGE genotypes. The results suggest that the Campylobacter strains isolated from the early stage of the production were susceptible to tetracycline, but they were subsequently displaced by tet-resistant Campylobacter. 相似文献