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应用质粒图谱分析大肠埃希氏菌毒力差异的研究 总被引：4，自引：0，他引：4
对1株无致病性鸡源大肠埃希氏菌（O21）和5株已知毒力强弱的大肠埃希氏菌（O2、O1、X2、148和O78）进行质粒图谱（PP）分析。结果表明这6个菌株属于不同的6种质粒谱型，其毒力的强弱与其所携带质粒的数量和大小有关，但不同毒力株间也有少数几条相同大小的谱带，试验结果表明PP图谱法可作为大肠埃希氏菌分型和鉴别其毒力的间接分子生物学方法。 相似文献
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为探明仔猪细菌性腹泻肠道致病性大肠埃希氏菌和沙门氏菌流行的血清型、耐药表型及耐药基因型,本试验采集了贵州省5个地（州）市7个规模化养猪场的128份腹泻仔猪肠道样本,并对采集的样本进行了大肠埃希氏菌和沙门氏菌分离与鉴定,通过动物试验鉴定菌株的致病性,利用血清学方法鉴定其血清型,并通过药敏纸片法对主要致病菌进行耐药性研究,采用PCR技术检测各致病菌株耐药相关基因,分析细菌耐药表型和耐药基因型相关性。结果显示,本研究共分离鉴定到78株致病性大肠埃希氏菌与21株沙门氏菌,致病性大肠埃希氏菌血清型以O138、O87为主,沙门氏菌血清型以鼠伤寒沙门氏菌、肠炎沙门氏菌居多;药敏试验结果表明,本试验分离到的78株致病性大肠埃希氏菌对β-内酰胺类药物耐药率达80%以上,对其他种类的抗菌药耐药率均超过40%,分离鉴定的21株沙门氏菌对氨基糖苷类药物耐药率达50%以上,对其他种类的抗菌药耐药率均达20%以上;本试验分离鉴定的致病性大肠埃希氏菌共检出12种耐药相关基因,沙门氏菌共检出10种耐药相关基因,两种细菌耐药基因型与耐药表型符合率均达60%以上,且均为多重耐药。本研究为仔猪腹泻的综合防控提供了理论依据。 相似文献
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为了解宁夏地区规模化牛场犊牛腹泻源产超广谱β-内酰胺酶(ESBLs)大肠埃希氏菌耐药和遗传变异情况，对分离得到的17株产ESBLs大肠埃希氏菌，采用K-B纸片法进行药物敏感性试验，采用脉冲场凝胶电泳进行分型研究。结果显示，17株分离株对试验药物多数呈耐药状态，对头孢拉定、四环素、阿莫西林3种药物完全耐药，其他药物的耐药性为70.24%～94.12%,只对阿米卡星、诺氟沙星敏感性较高。分离菌株可分为17个PFGE型，菌株间的相似度为67.5%～85.7%,无优势带型。宁夏地区犊牛腹泻源产ESBLs大肠埃希氏菌耐药性严重，遗传差异较大，但不同菌株存在着不同程度的亲缘关系。 相似文献
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《动物医学进展》2021,42(6)
某牧场母牛产后发生体温升高、呼吸困难、乳汁中带血等症状,发病牛衰竭死亡,针对此种情况,采取病死牛的心脏、肝脏、子宫、乳汁等进行病原菌的分离和纯化,对其进行16S rRNA测序鉴定,毒力基因检测,致病性试验和药敏试验,还对牛的饮用水进行细菌数量的检测。结果显示,致病菌为大肠埃希氏菌、沙雷氏菌和产色葡萄球菌。在毒力基因检测试验中,从病料及饮用水分离出的20株大肠埃希氏菌中有3株携有毒力基因hlyA、STb、F17。致病性试验显示,在分离出的8株大肠埃希氏菌中4株为致病性大肠埃希氏菌,分离出的沙雷氏菌和产色葡萄球菌对小鼠均有致病力,可使部分小鼠死亡。药敏试验显示,分离出的致病性大肠埃希氏菌普遍对庆大霉素、环丙沙星、美罗培南敏感;黏质沙雷氏菌对庆大霉素、环丙沙星、美罗培南等5种药物敏感,产色葡萄球菌对庆大霉素等11种药物敏感。水质检验显示,该牛场牛的饮用水中细菌总数和大肠埃希氏菌数量均严重超出国家标准。研究结果对预防、治疗奶牛产后败血症都具有指导意义。 相似文献
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以往防治鸡大肠杆菌病主要采用抗生素,由于大肠埃希氏菌抗药性的产生,药物使用剂量越来越大,造成禽产品中抗生素的残留,危害人类健康,我们从1987年至今,在内蒙古中西部地区分离到鸡源致病性大肠埃希氏菌273株。经详细鉴定发现内蒙古中西部地区流行的鸡致病性大肠埃希氏菌主要是O78、O1、O2、O8、O9、和O132。因此,我们以这6株菌作为制苗菌株,进行了鸡大肠杆菌病多价油乳剂灭活苗的研制。1材料与方法1.1材料制苗菌株为内蒙古中西部地区分离的鸡源致病性大肠埃希氏菌O78、O1、O2、O8、O9和O132;肉汤培养基、甘油琼脂培养基按文献介绍的方… 相似文献
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从一起因呕吐、食欲减退而死亡的鹈鹕的内脏中分离到3株细菌,其培养性状及生化特性基本一致,经VITEK-32鉴定均为大肠埃希氏菌。动物试验表明,分离株对小鼠均具有较强的致病性,且从死亡鼠内脏中回收到相应细菌结果表明,分离到的菌株为致病性大肠埃希氏菌。 相似文献
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Chinen I Otero JL Miliwebsky ES Roldán ML Baschkier A Chillemi GM Nóboli C Frizzo L Rivas M 《Research in veterinary science》2003,74(3):283-286
Four hundred and twenty-two calves were examined for intestinal carriage of Shiga toxin-producing Escherichia coli O157:H7 using conventional plating. Two (0.5%) E. coli O157 were recovered. They were compared with 96 Argentine strains of different origin by pulsed-field gel electrophoresis, phage typing and PCR-RFLP of stx2 genes. One strain isolated from a calf, was closely related with 18 strains of clinical origin. 相似文献
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Hiroi M Yamazaki F Harada T Takahashi N Iida N Noda Y Yagi M Nishio T Kanda T Kawamori F Sugiyama K Masuda T Hara-Kudo Y Ohashi N 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2012,74(2):189-195
To evaluate the diversity of extended-spectrum β-lactamases (ESBL) genes among food-producing animals, 48 isolates of ESBL-producing Escherichia coli isolates were obtained from rectal samples of broilers, layers, beef cattle and pigs, at the slaughterhouse level. ESBL-carrying E. coli were isolated from 60.0% of individual broiler rectal samples, 5.9% of layers, 12.5% of beef cattle and 3% of pigs. One ESBL-producing Klebsiella pneumoniae was isolated from a broiler. The ESBL-positive E. coli isolates from broilers harbored various ESBL genes: bla (SHV-12), bla(CTX-M-2), bla(CTX-M-14), bla(CTX-M-15) and bla(CTX-M-44). The plasmid DNAs were analyzed by restriction patterns. Homogeneous band patterns were yielded in those of K. pneumoniae and E. coli isolates harboring the bla(CTX-M-2) gene from different farms. No genetic relation between the 2 CTX-M-14 ESBL-producing strains was found by pulsed-field gel electrophoresis, although 2 plasmids in these strains, obtained from different broiler farms, were similar to each other. This study provides evidence that the proliferation of CTX-M-producing E. coli is due to the growth of indigenous CTX-M-producing strains and the possible emergence of strains that acquired CTX-M genes by horizontal transfer in different broiler farms. CTX-M-producing coliforms in broilers should be controlled due to the critical importance of cephalosporins and the zoonotic potential of ESBL-producing bacteria. 相似文献
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采用固定黏蛋白模型,结合细菌同位素γ-32P-ATP标记法,探讨犬源肠球菌E01和E16、犬源大肠杆菌DE、鸡源德氏乳酸杆菌德氏亚种N11及猪源约氏乳杆菌JJB3和致病性大肠杆菌ATCC25922和CVCC2060对本地犬各肠段黏蛋白的黏附能力;肠球菌E16和大肠杆菌DE各自的黏附机制;2株犬源肠球菌以3种作用方式（排斥、竞争、取代）对2株致病性大肠杆菌的黏附抑制作用。结果表明,各测试菌株在犬各肠段黏蛋白模型上的黏附值均不同;2株犬源肠球菌和1株猪源乳酸菌的黏附性能显著优于犬源大肠杆菌DE和2株致病性大肠杆菌;乳酸肠球菌E16表面的碳水化合物结构和糖蛋白参与了黏附过程,大肠杆菌菌体蛋白在黏附过程中发挥一定作用;乳酸肠球菌的黏附受体对高碘酸钠和胰蛋白酶处理不敏感,大肠杆菌DE的黏附受体具有碳水化合物结构,可能为糖蛋白类物质;在肠球菌与病源菌的3种作用方式中,取代和排斥方式对病原菌黏附抑制效果较好。这表明2株犬源肠球菌具开发为益生菌的潜力。 相似文献
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Platell JL Cobbold RN Johnson JR Clabots CR Trott DJ 《Comparative immunology, microbiology and infectious diseases》2012,35(4):319-324
Antimicrobial-resistant extraintestinal pathogenic Escherichia coli (ExPEC) impact both human and veterinary medicine. One ExPEC clonal group that has become increasingly multidrug-resistant is serotype O15:K52:H1. Accordingly, we sought O15:K52:H1 strains among fluoroquinolone-resistant (FQ(r)) E. coli clinical isolates from humans (n=582) and dogs (n=120) in Australia. The phylogenetic group D isolates (267/702; 38%) were screened for O15:K52:H1-specific single-nucleotide polymorphisms (SNPs) in fumC and the O15 rfb variant. The 34 so-identified O15:K52:H1 isolates (33 human, 1 canine) underwent antimicrobial susceptibility profiling, virulence genotyping, and macrorestriction profiling. Although susceptibility profiles varied, the 34 isolates were closely related by pulsed-field gel electrophoresis and exhibited typical O15:K52:H1-associated virulence profiles (complete pap operon, F16 papA allele, papG allele II, iha, fimH, sat, fyuA, iutA, kpsMII, ompT). The canine isolate closely resembled human isolates. Thus, O15:K52:H1 strains contribute to the FQ(r) ExPEC population in Australia and may potentially be transferred between humans and dogs. 相似文献
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Between May and September 2004, fecal samples from various wildlife species admitted to two rehabilitation centers in Ohio were cultured for Salmonella enterica and Escherichia coli O157:H7. Eight of 71 (11%) samples, including specimens from three opossums (Didelphis virginiana), two gray squirrels (Sciurus carolinensis), a woodchuck (Marmota monax), a Harris hawk (Parabuteo unicinctus), and a screech owl (Otus asio) tested positive for Salmonella serovars Braenderup, Senftenberg, Oranienburg, and Kentucky. The Salmonella Oranienburg isolates were indistinguishable by pulsed-field gel electrophoresis. Most isolates were susceptible to commonly used antibiotics; however, the Salmonella Kentucky isolate was resistant to multiple beta-lactam antibiotics (amoxicillin/clavulanic acid and ampicillin), cefoxitin, and ceftiofur, a third-generation cephalosporin. Escherichia coli O157:H7 was not isolated from any sample. Transmission of Salmonella from wildlife may occur between animals at rehabilitation centers. 相似文献
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Pyometra (uterine inflammation with accumulation of pus in the uterus) is regarded as one of the most common illnesses in bitches. The ethiology and pathogenesis are complex with both hormonal and bacterial elements. The bacteria most frequently isolated from the uterine content is Escherichia coli.In this study, 84 E. coli strains from the uteri of 70 bitches suffering from the disease were examined and their DNA-profiles compared by restriction enzyme analysis and pulsed-field gel electrophoresis (PFGE). Through variations in DNA-profiles of the E. coli isolates, this study indicates that pyometra is caused by E. coli originating from the normal flora of the dogs and not by certain clones spread between animals.E. coli strains from the urinary bladder and the uterus of six of the bitches suffering from simultaneous urinary tract infection and pyometra were examined and compared as above. The DNA-profiles of the isolates from each of the six bitches were 100% identical. This study supports the theory suggesting that in cases of simultaneous urinary tract infection and E. coli pyometra, the urinary tract and uterus are infected with the same bacterial strain.To evaluate whether the uterus was infected with a single clone of E. coli or if multiple clones were present, eight to 16 colonies of E. coli isolated from pyometra samples from a further 10 bitches were examined. All bacterial colonies from the culture of the same bitch showed identical DNA-profiles.In 14 of the 70 bitches, two macroscopically different but biochemically identical E. coli colony types were isolated. The two colony types from the same bitch proved to have identical DNA-profiles in 13 cases and almost identical in the remaining bitch. 相似文献
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Guth BE Chinen I Miliwebsky E Cerqueira AM Chillemi G Andrade JR Baschkier A Rivas M 《Veterinary microbiology》2003,92(4):335-349
Shiga toxin (Stx)-producing Escherichia coli (STEC) strains isolated from animals and food in Argentina (n=44) and Brazil (n=20) were examined and compared in regard to their phenotypic and genotypic characteristics to evaluate their pathogenic potential. The clonal relatedness of STEC O157 isolates (n=22) was established by phage typing (PT) and pulsed-field gel electrophoresis (PFGE). All O157 strains studied carried eae and enterohemorrhagic E. coli (EHEC)-hly sequences. In Argentina, these strains occurred both in cattle and meat, and 50% of them carried stx2/stx2vh-a genes, whereas in Brazil the O157 strains were isolated from animals, and most harbored the stx2vh-a sequence. At least 13 different O:H serotypes were identified among the non-O157 strains studied, with serotype O113:H21 being found in both countries. All but one non-O157 strains did not carry eae gene, but EHEC-hlyA gene was found in 85.7% of them, and the stx2 genotype was also more prevalent in Argentina than in Brazil (P<0.01), where stx1 alone or in association was most common (68.8%). One STEC strain isolated from a calf in Brazil harbored the new variant referred to as stx2-NV206. PFGE analysis showed that STEC O157 strains were grouped in four clusters. One Brazilian strain was considered possibly related (> or =80%) to Argentinean strains of cluster I. Differences in the pathogenic potential, especially in regard to serotypes and stx genotypes, were observed among the STEC strains recovered from animals and food in both countries. 相似文献
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In the autumn of 1995 the first outbreaks of enterohemorrhagic Escherichia coli O157:H7 including ca 100 human cases were reported in Sweden. From outbreaks in other countries it is known that cattle may carry these bacteria and in many cases is the source of infection. Therefore, the present study was performed to survey the Swedish bovine population for the presence of verotoxin-producing E. coli (VTEC) of serotype O157:H7. Individual faecal samples were collected at the 16 main Swedish abattoirs from April 1996 to August 1997. Of 3071 faecal samples, VTEC O157 were found in 37 samples indicating a prevalence of 1.2% (CI95% 0.8-1.6). All 37 isolates carried genes encoding for verotoxin (VTI and/or VT2), intimin, EHEC-haemolysin and flagellin H7 as determined by PCR. Another 3 strains were of serotype O157:H7 but did not produce verotoxins. The 37 VTEC O157:H7 strains were further characterised by phage typing and pulsed-field gel electrophoresis. The results clearly show that VTEC O157:H7 is established in the Swedish bovine population and indicate that the prevalence of cattle carrying VTEC O157:H7 is correlated to the overall geographical distribution of cattle in Sweden. Results of this study have formed the basis for specific measures recommended to Swedish cattle farmers, and furthermore, a permanent monitoring programme was launched for VTEC O157:H7 in Swedish cattle at slaughter. 相似文献
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Eight strains of Taylorella equigenitalis were identified by a polymerase chain reaction using a primer pair specific to the 16S rDNA of T equigenitalis. These eight strains were chosen because they had previously been shown to represent eight distinct genotypes by pulsed-field gel electrophoresis analysis after separate digestion of the genomic DNA with ApaI or NotI. The eight strains could be classified into six or seven types by random amplified polymorphic DNA analysis using different kinds of primers. Amplified rDNA restriction analysis after separate digestion with five restriction enzymes, including AluI and MboI, of the 1,500 bp fragments of rDNA amplified by polymerase chain reaction did not discriminate the genomic variations among the eight strains of T equigenitalis. Thus, pulsed-field gel electrophoresis was shown to discriminate these eight organisms better than random amplified polymorphic DNA analysis, while amplified rDNA restriction analysis was found to be unsuitable for subtyping T equigenitalis. 相似文献