Characterization of integrons in multiple antimicrobial resistant Escherichia coli isolates from bovine endometritis

To assess the prevalence of antimicrobial resistance and three classes of integrons in Escherichia coli (E. coli) strains (n = 57) isolated from bovine endometritis in Inner Mongolia of China, antimicrobial susceptibility and the presence of three types of integrons were characterized. Most isolates were susceptible to ceftiofur, furazolidone, ciprofloxacin and enrofloxacin, while 57 isolates were all resistant to sulfamethoxydiazine and trimethoprim. High resistant incidence rates were exhibited to sulfadiazine, tetracycline, oxytetracycline, cefazolin, chloramphenicol. Forty-six of 57 E. coli strains were resistant to above 10 antibiotics (80.70%). The integrase gene and gene cassettes of integrons were amplified by PCR. DNA sequencing and analysis were used to identify the genetic content of the integron-variable regions. Neither class II nor class III integron was detected, while 36.8% (n = 21) of the isolates were positive for the presence of intI1 gene. Analysis of gene cassettes revealed that six gene cassettes were found, which encoded resistance to trimethoprim (dhfr, dhfrI, dfrA17) and aminoglycosides (aadA1, aadA2, aadA5). Among them, the gene cassette array dfrA17–aadA5 was found most prevalent (66.7%). The resistance profile of positive-integron isolates was relatively broad and they were resistant to more than eight antimicrobials (n ? 8). The correlation analysis revealed the incidence of integrons among the isolates were related to the multiple antibiotic resistance profile, indicating integrons play an important role in the dissemination and spread of the antimicrobial resistant strains.     

腹泻犊牛大肠杆菌血清型抗菌药物耐药性耐药基因鉴定及脉冲场凝胶电泳分型

本试验对2011年由哈尔滨周边地区采集的263份腹泻犊牛的肛拭子进行大肠杆菌的分离鉴定,并通过小鼠致病性试验鉴定出26株致病性大肠杆菌,用Kirby-Bauer法开展对12种药物的药敏试验,PCR检测了我国流行广泛的9种耐药基因,脉冲场凝胶电泳(PFGE)进行基因分型.检测结果,该地区主要流行的致病性血清型为O26、O78和O15,所有致病性菌株都存在多重耐药现象,其中阿莫西林-棒酸、链霉素、四环素、复方新诺明的耐药率达到了100％,共检出7种耐药基因,检出率符合我国平均分布趋势,且检出率基本上与耐药表型相一致.PFGE结果显示,该地区存在致病性菌株的克隆传播.     

The prevalence of verocytotoxin-producing Escherichia coli and antimicrobial resistance patterns of nonverocytotoxin-producing Escherichia coli and Salmonella in Ontario broiler chickens.

The prevalence of verocytotoxin-producing Escherichia coli and Salmonella in Ontario broiler chickens was determined by culturing cloacal samples from 500 individual birds selected from 50 poultry farms. Resistance to antimicrobials was determined for each of the isolates. In addition, abattoir and farm-level management data were obtained to evaluate variables that may be considered risk factors for infection. The variables selected included: Percentage of birds condemned at slaughter, percentage of birds dead-on-arrival, bird weight, truck number, farm size, hatchery source, litter source and type, feed source, mortality levels, type of water drinker, water sanitization, down time, barn clean out and history of antibiotic treatment. None of the cloacal samples revealed the presence of verocytotoxin-producing E. coli, though 19/500 (3.8%) contained Salmonella organisms. Nine different Salmonella serotypes were isolated; the most common being S. hadar, S. heidelberg and S. mbandaka. Resistance to tetracycline and streptomycin was common among Salmonella (63%) and E. coli (25.2%) isolates. Resistance to two or more antimicrobials occurred in 420/500 (84%) of the E. coli isolates. No statistically significant associations between abattoir or farm-level management variables and the Salmonella-status of farms were demonstrated.     

Detection and characterization of Shiga toxin-producing Escherichia coli in captive non-domestic mammals

Shiga toxin producing-Escherichia coli (STEC) is an important emerging pathogen, and ruminants are recognized as their main natural reservoir. The aim of this work was to establish the frequency of STEC in non-domestic mammals of the Zoo and Botanical Garden of La Plata City, Argentina, and to pheno-genotypically characterize STEC isolates. By polymerase chain reaction (PCR), Shiga toxin (stx) gene sequences were detected in 50.8% of 65 fecal samples. Twenty-five STEC strains were isolated from 38.5% of the Zoo's animals. Ten species of order Cetartiodactyla and one species of order Rodentia were recognized as new STEC carriers. STEC strains belonged to 7 different serotypes including new serotypes O12:H25 and O13:H6. Serotype O146:H28, previously associated with human infections, represented 24% of STEC isolates. The most frequent Shiga toxin identified were type 1c and type 2c. Nineteen strains were positive for iha gene, 8 strains were positive for ehxA gene. Moreover, all strains were positive for lpfAO113 and negative for rfbO157, eae, saa, lpfAO157/OI-141, lpfAO157/OI-154, efa1, and toxB genes. Results obtained by XbaI-pulsed-field gel electrophoresis (XbaI-PFGE) confirmed the transmission of STEC strains among different animal species and suborders. In addition, we observed a potential association between STEC-harboring animal and factors such as belonging to order Cetartiodactyla, living in a pit, and belonging to a non-autochthonous species. This is the first work developed with zoological mammals and STEC in Argentina.     

Comparison of antimicrobial resistance in generic Escherichia coli and Salmonella spp. cultured from identical fecal samples in finishing swine

The objectives of this study were to investigate the associations between antimicrobial resistance patterns in generic Escherichia coli and Salmonella spp. isolates recovered from identical pen pooled fecal samples, and to evaluate potential clustering of multiple isolates of these organisms within identical fecal samples. Up to 5 generic E. coli (n = 922 isolates) and Salmonella spp. (n = 922 isolates) isolates were obtained from each of 188 pen pooled fecal samples that had been collected from 45 finishing swine farms in Alberta in 2000, and tested for susceptibility to 15 antimicrobials. No isolates of either organism were resistant to 3rd generation cephalosporins or fluoroquinolones, which in Canada are considered antimicrobials of very high importance to human health. Approximately twice as many generic E. coli isolates as Salmonella spp. isolates were resistant to at least 1 antimicrobial. In addition, E. coli isolates showed more multidrug-resistance patterns. No significant association was observed between the resistance phenotypes of Salmonella spp. and E. coli at the fecal sample level. More clustering at the sample level was observed for proportions of antimicrobial resistance (AMR) in Salmonella spp. isolates than E. coli indicating that in future studies it might be sufficient to test fewer than 5 Salmonella spp. isolates per sample.     

Molecular and biological characterization of vascular endothelial growth factor of parapoxviruses isolated from wild Japanese serows (Capricornis crispus)

Contagious dermatitis in domestic and wild ruminants caused by parapoxvirus (PPV) occurs worldwide. Although PPV infections appear in cattle, sheep and goats, the papular, nodular, pustular and ulcerated skin lesions of wild Japanese serows (Capricornis crispus) are significantly more severe than those of other animals. To determine the factors involved in the severity of these skin lesions, we compared the molecular characteristics of 4 PPV isolates from Japanese serows and 2 isolates from sheep in Japan, and also investigated the biological properties of primary endothelial cells from different host species. All of the 6 Japanese isolates harbored a vascular endothelial growth factor (VEGF) gene, which is known as one of the virulence factors of PPVs. Three different amino acid sequences of viral VEGF were identified and the sequence was identical among the 4 isolates from the Japanese serows. The temporal expression pattern of viral VEGF mRNA was almost the same among 3 isolates encoding the 3 different VEGFs in infected cells from different host species. Recombinant forms of the 3 different VEGFs showed the ability to induce vascular permeability and endothelial cell proliferation. Primary endothelial cells from Japanese serows were most responsive to recombinant viral VEGF compared to cells from cattle, sheep, and goats. These results suggest that not only the biological activity of viral VEGF but also the high responsiveness to viral VEGF of endothelial cells might be involved in the severe proliferative skin lesions induced by PPV infection in Japanese serows.     

Mycoplasma ovis detected in free-living Japanese serows, Capricornis crispus

Nineteen blood samples collected from free-ranging wild Japanese serows, Capricornis crispus, between 2006 and 2008 in Iwate prefecture were examined for the hemoplasma infection by real-time PCR targeting the 16S rRNA gene. Five (26.3%) out of the 19 samples were positive in real-time PCR with an average melting temperature at 85.18 °C. The positive samples in the real-time PCR were reconfirmed by conventional PCR, and one of them was successful for direct DNA sequencing. The nucleotide sequence of the 16S rRNA gene of the representative stain was identical to that of Mycoplasma ovis. This was the first demonstration of hemotropic mycoplasma infections among the free-living Japanese serows in Japan.     

Cecal bacterial communities in wild Japanese rock ptarmigans and captive Svalbard rock ptarmigans

Preservation of indigenous gastrointestinal microbiota is deemed to be critical for successful captivebreeding of endangered wild animals, yet its biology is poorly understood. Here, we investigated cecalbacterial communities in wild Japanese rock ptarmigans (Lagopus muta japonica) and comparedthem with those in Svalbard rock ptarmigans (L. m. hyperborea) in captivity. Ultra-deepsequencing of 16S rRNA gene indicated that the community structure of cecal microbiota in wild rock ptarmiganswas remarkably different from that in captive Svalbard rock ptarmigans. Fundamental differences betweenbacterial communities in the two groups of birds were detected at the phylum level. Firmicutes,Actinobacteria, Bacteroidetes and Synergistetes were the major phyla detected in wild Japanese rockptarmigans, whereas Firmicutes alone occupied more than 80% of abundance in captive Svalbard rock ptarmigans.Furthermore, unclassified genera of Coriobacteriaceae, Synergistaceae, Bacteroidaceae, Actinomycetaceae,Veillonellaceae and Clostridiales were the major taxa detected in wild individuals, whereas in zoo-rearedbirds, major genera were Ruminococcus, Blautia, Faecalibacterium andAkkermansia. Zoo-reared birds seemed to lack almost all rock ptarmigan-specific bacteria intheir intestine, which may explain the relatively high rate of pathogenic infections affecting them. We showevidence that preservation and reconstitution of indigenous cecal microflora are critical for successfulex situ conservation and future re-introduction plan for the Japanese rock ptarmigan.     

西藏牦牛大肠杆菌耐药性调查

对西藏部分牧区牛源大肠杆菌进行耐药性考察,在对发病牦牛进行致病性大肠杆菌分离鉴定、纯化培养的基础上,采用美国NCCLS规定的纸片扩散法进行药敏试验,结果表明,分离培养的10株牛源大肠杆菌对头孢氨苄、头孢唑林、卡那霉素、阿米卡星、磷霉素、喹诺酮类药物的敏感率均为100%,可为临床合理选药、有效治疗牦牛大肠杆菌病提供依据。     

鸡致病性大肠埃希氏菌耐氧氟沙星质粒的检测

对分离的２０株鸡致病性大肠杆菌进行了药敏实验,筛选出１３株耐氧量沙星（ＯＦＬ）的菌株。经质粒提取,电泳和转化实验,获得一株 粒倡导 的耐煌多重耐药菌株。该菌株对参试的１１种抗生素全部耐药菌株其中ＯＦＬＭＩＣＭ〉５０μｇ／ｍｌ。该菌Ｒ质粒电泳可见６条带,转化大肠杆菌ＤＨ５α,在２０μｇ／ｍｌＯＦＬ的ＬＢ学板筛选得到的转化子,该转化子含有原菌株中２．７ｋｂ的质粒,获得原菌株部分耐药性。连续转化５次,均     

Detection and characterisation of Shiga toxin-producing Escherichia coli other than Escherichia coli O157:H7 in wild ruminants

Shiga toxin-producing Escherichia coli (STEC) are an important group of emerging pathogens, with ruminants recognised as their main natural reservoir. The aim of this work was to establish the prevalence of non-O157 STEC in free-ranging wild ruminants in the Extremadura region of Spain and to characterise them phenogenotypically. Faecal samples were collected from 243 wild ruminants, including Cervus elaphus, Capreolus capreolus, Dama dama and Ovis musimon and were examined for STEC using both phenotypic (Vero cells) and genotypic (PCR and PFGE) methods.Shiga toxin-producing Escherichia coli were isolated from 58 (23.9%) of the samples and a total of 65 isolates were characterised. A PCR method indicated that 11 (16.9%) strains carried the stx₁ gene, 44 (67.7%) carried the stx₂ gene and 10 (15.4%) carried both these genes. The ehxA gene was detected in 37 (57%) of the isolates but none contained either the eae or saa genes. The isolates were from a total of 12 ‘O’ serogroups, although 80% were restricted to the O2, O8, O128, O146, O166 and O174 serogroups. The most commonly isolated STEC bacteria, which were from the O146 serogroup, exhibited a high degree of polymorphism as indicated by PFGE. Shiga toxin-producing Escherichia coli isolates of serogroups O20, O25, O166, O171, O174 and O176 had not previously been found in wild ruminants. This is the first study to confirm that wild ruminants in Spain are a reservoir of STEC and are thus a potential source of human infection.     

Molecular basis of quinolone resistance in Escherichia coli from wild birds

Nine quinolone resistant (minimal inhibitory concentration [MIC] was > 32 microg/mL for nalidixic acid, > 1 microg/mL for ciprofloxacin) isolates of Escherichia coli have been found in wild birds with septicemia. All of the isolates were aerobactin positive. The mechanisms of resistance were characterised by sequencing the quinolone resistance-determining region (QRDR) of the gyrA, gyrB, parC, and parE genes. Sequence analysis of the gyrA gene in all isolates identified only 1 nucleotide substitution at codon Serine-83 for Leucine-83. Sequence analysis of the gyrB, parC, and parE QRDR genes revealed no mutations in any of the isolates. This study was conducted to determine the importance of these genes in the susceptibility of E. coli strains isolated from wild birds to quinolones.