Occurrence of the temperature-sensitive hemagglutinin among avian Escherichia coli

In this study, we determined the occurrence of the tsh gene among 305 Escherichia coli isolates from chickens by means of the polymerase chain reaction and agglutination of chicken erythrocytes; 200 of those isolates were obtained from chickens with colisepticemia, 52 isolates were from lesions of cellulitis, and 53 were from feces of normal chickens. The tsh gene was found in 79 (39.5%) isolates from colisepticemia, in 10 (19%) cellulitis-derived E. coli isolates, and in two (3.8%) fecal isolates. Among the tsh+ strains, 68 (86%) isolates from colisepticemia and nine (90%) from cellulitis agglutinated chicken erythrocytes in the presence of mannose, after growing the strains on colonization factor antigen agar plates at 26 C, which confirms a correlation between mannose-resistant hemagglutination and expression of hemagglutinin Tsh. These results show, for the first time, the presence of the gene tsh in cellulitis-derived E. coli isolates; the high frequency of this gene among avian pathogenic E. coli isolates in Brazil indicates that its putative role as a virulence factor should be studied more thoroughly.     

Use of epidemiologic markers to identify the source of Escherichia coli infections in poultry

During an epidemiologic study of poultry colisepticemia on 2 Saudi Arabian poultry broiler farms, Escherichia coli was isolated from 101 (40.4%) of the 250 specimens examined. The antigenic structure and the drug resistance pattern of 65.4% of the E coli isolates from different sources were used as epidemiologic markers to trace the source of the infection. The predominant E coli serotypes involved in infections of 2 poultry broiler progeny farms were 033:H4 (51.8%) and 078:H- (19.6%) that had the following respective drug resistance patterns: furazolidone-streptomycin-sulfathiazole and streptomycin-sulfathiazole-tetracycline. Escherichia coli strains with typical epidemiologic markers were isolated from various sources on a broiler breeding farm, but not from well waters of the infected progeny farm. Three other E coli serotypes (045:H10[14.3%], 0119:H27[1.8%], and 0145:H25[1.8%]) were involved in poultry infection, but to a lesser extent. These 3 serotypes were multiply resistant against 5 to 6 of the antimicrobials evaluated.     

Characterization of extraintestinal Escherichia coli isolated from a peacock (Pavo cristatus) with colisepticemia

Extraintestinal infections by avian pathogenic strains of Escherichia coli (APEC) are commonly reported in poultry, but there is little information on infections by APEC in other bird species. Here we report on the characterization of extraintestinal E. coli isolated from a domesticated peacock, from the south of Brazil, that died of colisepticemia. Necropsy examination revealed congested liver, hypertrophied kidneys, peritonitis, severe typhlitis suggestive of coligranuloma, pneumonia, and airsacculitis--typical signs of colisepticemia. The isolates from lungs, kidney, heart, intestine, liver, and bone marrow all harbored the same virulence-associated factors (iucD, colV, iss, mat, fimC, ompA, traT crl, csgA vgrG, and hcp), yielded the same band pattern in amplified ribosomal DNA restriction analysis, and were allocated to the Escherichia coli Reference Collection group B1. The isolates were resistant to bacitracin, trimethoprim, and tetracycline, but displayed slight differences in their resistance to other antimicrobials. The isolates also differed in their virulence in 1-day-old chickens, but none displayed high virulence in vivo. We conclude that the peacock died of colisepticemia after it was infected with an extraintestinal E. coli strain of low virulence that nevertheless harbored virulence factors generally associated with APEC. This study represents the first characterization of an APEC isolated from a nonpoultry bird species.     

Congo red medium to distinguish between invasive and non-invasive Escherichia coli pathogenic for poultry

In the course of our molecular studies of virulence factors associated with invasive avian Escherichia coli infections, it was first necessary to distinguish between common E. coli and those that cause septicemia in poultry. We found a direct correlation between the ability of clinical isolates of E. coli to bind Congo red dye (CR) and their ability to cause septicemic infection in chickens. This finding was supported by bacteriological studies of 30 broiler flocks (26 sick and 4 healthy) and by virulence studies in chickens and mice. All 144 isolates of E. coli from internal tissues of diseased birds were determined to be CR-positive (red colonies). Congo-red-positive E. coli colonies were isolated from air sacs, pericardium, liver, lung, joint fluid, and heart blood of chickens with lesions of colisepticemia. In contrast, of 170 E. coli isolates from the poultry house environment and from the trachea and cloaca of healthy birds, more than half were CR-negative (white colonies). No CR-negative (white) E. coli colonies were found in internal organs from birds with typical lesions of colisepticemia. We feel that these preliminary findings suggest that the CR dye binding could be used as a phenotypic marker to distinguish between invasive and noninvasive isolates.     

Virulence factors associated with Escherichia coli present in a commercially produced competitive exclusion product

In this study, we assessed the pathogenic potential of Escherichia coli associated with a commercial competitive exclusion (CE) product by examining the phenotypic characteristics associated with E. coli virulent for humans and domestic animals. Most E. coli isolates were capable of proliferating in iron-deplete chicken sera. Interestingly, none of the E. coli isolates from the commercial CE product contained the bacterial adhesin Tsh characteristic of avian pathogenic E. coli associated with airsacculitis and colisepticemia. In terms of virulence potential for humans, most E. coli isolates (78%) were sensitive to killing by 12.5% human sera. Because of their sensitivity to human sera, the E. coli in the CE product are not likely to cause a serious systemic infection in humans and, therefore, do not present a risk of causing septicemia in humans. Because these isolates also lack the gene tsh, they are also less likely to cause systemic disease or airsacculitis in poultry than pathogenic strains commonly isolated from diseased birds.     

Antibiotic resistance among indicator bacteria isolated from healthy pigs in New Zealand

AIM: To determine the resistance to antibiotics among the indictor bacteria, Escherichia coli and Enterococcus spp, isolated from the faeces of healthy pigs on three conventional pig farms and one organic farm in the North Island of New Zealand. METHODS: Faecal samples, collected at intervals between March and October 2001, were plated onto MacConkey agar and Slanetz-Bartley agar and examined after 1-3 days incubation for colonies resembling E. coli and Enterococcus spp, respectively. Typical colonies were subcultured for further identification and storage. The isolates were tested for antibiotic resistance, using disc diffusion, to ampicillin, gentamicin, streptomycin, and tetracycline. Escherichia coli isolates were also tested for resistance to ciprofloxacin, cotrimoxazole and neomycin. Enterococcus spp isolates were also tested for resistance to vancomycin, erythromycin and virginiamycin. RESULTS: A total of 296 E. coli and 273 Enterococcus spp isolates were obtained from the three conventional farms, and 79 E. coli and 80 Enterococcus spp isolates were obtained from the organic farm. All the E. coli isolates from both the conventional and organic pig farms were susceptible to ciprofloxacin, and all the Enterococcus spp isolates were susceptible to ampicillin, gentamicin and vancomycin. Isolates of E. coli from conventional pig farms were resistant to gentamicin (0.7%), neomycin (0.7%), ampicillin (2.7%), cotrimoxazole (11%), streptomycin (25%) and tetracycline (60%). Enterococcus spp isolates from the same farms were resistant to erythromycin (68%), tetracycline (66%), streptomycin (54%) and virginiamycin (49%). By contrast, for the organic pig farm 相似文献   

Antimicrobial susceptibility of Escherichia coli isolates from dairy farms using organic versus conventional production methods

OBJECTIVE: To compare antimicrobial susceptibility patterns of Escherichia coli isolates cultured from fecal samples from cows and calves on dairy farms that used organic (ie, no or severely limited antimicrobial use) versus conventional production methods. DESIGN: Cross-sectional study. SAMPLE POPULATION: Fecal samples from 10 cows and 10 calves on each of 30 organic dairy farms and 30 neighboring conventional dairy farms in Wisconsin. PROCEDURE: E. coli isolates obtained from the fecal samples were tested for susceptibility to 17 antimicrobials by means of a microbroth dilution test. Prevalence of antimicrobial resistance was compared between organic and conventional dairy farms. RESULTS: E. coli was isolated from 1,121 (94%) fecal samples. Farm type (organic vs conventional) and animal age (cow vs calf) were significantly associated with odds that E. coli isolates would be resistant to various antimicrobials. After controlling for age, logistic regression analyses indicated that isolates from conventional dairy farms had significantly higher rates of resistance to ampicillin, streptomycin, kanamycin, gentamicin, chloramphenicol, tetracycline, and sulfamethoxazole than did isolates from organic dairy farms. However, no significant differences were detected for the 10 other antimicrobials that were tested. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that compared with isolates from conventional dairy farms, E. coli isolates from organic dairy herds have significantly lower prevalences of resistance to 7 antimicrobials; however, prevalence of resistance was not significantly different for 10 other antimicrobials. Resistance was more common for isolates from calves than for isolates from adult dairy cows.     

Comparison of chicken plasma and guinea pig serum in a quantitative microtiter method of determining microbial complement resistance.

A quantitative microtiter method using chicken plasma is described for determining the degree of complement resistance or sensitivity of avian Escherichia coli isolates. Results obtained with the microtiter method using chicken plasma were compred with results obtained using commercially available standardized guinea pig serum as the source of complement. The test organisms consisted of five isolates of E. coli isolated from chickens. Three isolates were from flocks with colisepticemia; one was from a flock with omphalitis; and one isolate was a non-pathogenic control. Data were accumulated from the five avian E. coli isolates incubated at 35 C with either chicken plasma or guinea pig serum and with heat-inactivated chicken plasma or guinea pig serum. The microtiter results of the chicken plasma and guinea pig serum had a statistically positive correlation. The use of commercially available guinea pig serum in the test system will allow for standardization of this method.     

Characterization of Escherichia coli isolates from peritonitis lesions in commercial laying hens

Five clinically normal chickens from three farms (farm A, farm B, and farm C), for a total of 15 clinically normal chickens, were examined bacteriologically. In a similar manner, five dead chickens with lesions of peritonitis from each of the same three commercial egg-laying operations were selected for bacterial culturing. Escherichia coli were isolated from the cloaca in 14 of 15 healthy chickens and from all 15 chickens with peritonitis. Oviducts of normal chickens did not contain E. coli (0/15) whereas oviducts from 13 of 15 hens with peritonitis were positive for this pathogen. No lesions and no E. coli (0/15) were found in the peritoneal cavity of healthy hens, but peritonitis lesions from 13 of 15 dead chickens yielded E. coli. On farm A and farm B, a flock consisted of all chickens within a single house and all chickens in each flock were of the same age and same genetic strain. In flock 1 from farm A, all five E. coli isolates from the oviduct and all five isolates from the peritoneal cavity were serogrouped as O78; contained the virulence genes iroN, sitA, iutA, tsh, and iss; and belonged to phylogenetic group A. In flock 2 from farm B, all four E. coli isolates from the oviduct and all four isolates from the peritoneal cavity were serogrouped as O111; contained virulence genes iroN, sitA, iutA, traT, iss, and ompT; and belonged to phylogenetic group D. These data suggest that all chickens with peritonitis in a single flock on farms A and B were likely infected by the same E. coli strain. Escherichia coli isolates from the magnum and peritoneum had the same serogroup, virulence genotype, and phylogenetic group, which is consistent with an ascending infection from the oviduct to the peritoneal cavity.     

鸡源大肠杆菌的耐药性监测及生物学特性研究

从山东省诸城地区6个集约化养鸡场采集50只病死鸡病料,从中分离鉴定得到32株大肠杆菌,对其进行了12种常规药敏试验,发现分离的菌株有不同程度的耐药性。对14株优势血清型菌株进行了质粒DNA提取分析,然后用Hind Ⅲ限制性内切酶对质粒进行了酶切。结果表明,质粒的得率为100%,来源相同的菌株具有相同或相似的质粒图谱,也就是说它们有共同的起源;来源不同的菌株的质粒图谱一般不同,即有不同的起源。将血清型和质粒图谱比较,结果发现,同一血清型可以有不同的质粒图谱,不同血清型可以有相似的质粒图谱,血清型与质粒图谱没有直接的联系。     

Antimicrobial resistance of commensal Escherichia coli from dairy cattle associated with recent multi-resistant salmonellosis outbreaks

The use of antimicrobial drugs in livestock is suspected to contribute to bacterial antimicrobial resistance (AR) development. Dairy farms experiencing recent outbreaks of salmonellosis involving multi-resistant (MR) Salmonella strains were compared to control farms with respect to AR among bovine commensal E. coli isolates. For most antimicrobials tested, the percentage of AR E. coli isolated from salmonellosis-affected farms was significantly higher than that from control farms. Calf E. coli from both case and control farms had greater levels of AR than cow isolates. Commensal E. coli isolates from case farms and calves tended to more frequently be MR. These data are consistent with the existence of higher antimicrobial selection pressure on farms with recent salmonellosis outbreaks, however, the directionality of the relationship remains to be elucidated. An improved understanding of the epidemiology of AR bacteria in livestock production, both at the herd and molecular level, is essential to mitigate risk to public health and food safety.     

Studies on calf diarrhoea in Mozambique: prevalence of bacterial pathogens

The prevalence of diarrhoea in calves was investigated in 8 dairy farms in Mozambique at 4 occasions during 2 consecutive years. A total of 1241 calves up to 6 months of age were reared in the farms, and 63 (5%) of them had signs of diarrhoea. Two farms had an overall higher prevalence (13% and 21%) of diarrhoea. Faecal samples were collected from all diarrhoeal calves (n = 63) and from 330 healthy calves and analysed for Salmonella species, Campylobacter jejuni and enterotoxigenic Escherichia coli (ETEC). Salmonella spp. was isolated in only 2% of all calves. Campylobacter was isolated in 11% of all calves, irrespective of health condition, and was more frequent (25%) in one of the 2 diarrhoeal farms (p = 0.001). 80% of the isolates were identified as C. jejuni. No ETEC strains were detected among the 55 tested strains from diarrhoeal calves, but 22/55 (40%) strains from diarrhoeal calves and 14/88 (16%) strains from healthy calves carried the K99 adhesin (p = 0.001). 6,757 E. coli isolates were typed with a biochemical fingerprinting method (the PhenePlate) giving the same E. coli diversity in healthy and diarrhoeal calves. Thus it was concluded: i) the overall prevalence of diarrhoea was low, but 2 farms had a higher prevalence that could be due to an outbreak situation, ii) Salmonella did not seem to be associated with diarrhoea, iii) Campylobacter jejuni was common at one of the 2 diarrhoeal farms and iv) ETEC strains were not found, but K99 antigen was more prevalent in E. coli strains from diarrhoeal calves than from healthy, as well as more prevalent in one diarrhoeal farm.     

16S rRNA甲基化酶rmtB在猪肠道菌中的传播方式分析

为研究16S rRNA甲基化酶rmtB在猪肠道菌中的传播方式,用脉冲场凝胶电泳(PFGE)分别对来源于A、B猪场的48株rmtB阳性大肠埃希氏菌进行基因分型;用膜接合试验研究rmtB基因的水平传播方式;用微量稀释法对rmtB阳性菌及其接合子进行药敏试验。有45株rmtB阳性大肠埃希氏菌(45/48)能进行PFGE分型,可分为28种不同的PFGE基因型。其中来源于A猪场的20株菌可分为12种基因型,来源于B猪场的25株菌可分为16种基因型;46株rmtB阳性菌可以通过接合试验将rmtB基因及其介导的耐药性传递给受体菌E.coliC600和E.coli488 Rifr,接合频率从4.6×10-13~3.0×10-6不等,接合子对卡那霉素、阿米卡星、妥布霉素、西梭米星、萘替米星、庆大霉素6种二脱氧链霉胺类抗生素均高度耐药(MIC≥512μg/mL)。结果表明,rmtB基因位于接合性质粒上,该基因在两猪场的大肠埃希氏菌中既存在克隆传播,又存在水平传播,以水平传播为主。     

Antimicrobial resistance in generic fecal Escherichia coli from 29 beef farms in Ontario

The occurrence of antimicrobial resistance in generic Escherichia coli can serve as an indicator of the pool of resistance genes potentially available for transfer to pathogenic organisms. This study was conducted on 29 volunteer beef farms in Ontario to describe the prevalence and patterns of antimicrobial resistance in E. coli, and to describe changes in the prevalence of resistance during the feedlot stage of production. From the pooled fecal samples on 28 of the 29 farms, 31% of isolates from feedlots (n = 993) and 12% of isolates from cow-calf farms (n = 807) were resistant to one or more of 16 antimicrobials tested. No isolates were resistant to ceftriaxone, ciprofloxacin, gentamicin, or nalidixic acid, and < 1% of the pooled isolates were resistant to ceftiofur. Two percent of both feedlot and cow-calf isolates were resistant to > or = 5 antimicrobials. Cow-calf farms were at significantly lower risk than feedlots for having E. coli isolates that were resistant to streptomycin, sulfamethoxazole, and tetracycline. On average, the prevalence of sulfamethoxazole resistant E. coli isolates was significantly higher in calves than in cows. No resistance was observed to ceftriaxone or ciprofloxacin among isolates (n = 1265) obtained from individually sampled feedlot animals on 2 farms. Less than 1% of these isolates were resistant to gentamicin, nalidixic acid, and ceftiofur. From the individually sampled feedlot animals, resistance to streptomycin (on 1 farm), sulfamethoxazole, and tetracycline increased significantly from arrival to mid-point during the feeding period, and these levels persisted until market-readiness.     

Relationship of complement resistance and selected virulence factors in pathogenic avian Escherichia coli.

Complement resistance, antibiotic resistance profiles, and virulence profiles of 80 Escherichia coli isolates from the intestines of normal chickens (40 isolates) and chickens diagnosed as having colisepticemia (40 isolates) were compared. Differences were observed between the two groups for antibiotic resistance, siderophore production, presence of type 1 pili, complement resistance, motility, and size of plasmids. The systemic isolates were more likely to have siderophores and type 1 pili, and to be complement-resistant and motile than were the intestinal isolates. No differences between the two groups were observed for colicin production. Further comparison of the 10 most complement-resistant isolates from the systemic group and 10 most complement-sensitive isolates from the intestinal group revealed a correlation between an isolate's resistance to complement and its ability to kill embryos, express type 1 pili, and be motile. Virulence of avian E. coli strains appears to be correlated with complement resistance and the interaction of this resistance with the ability to produce type 1 pili and be motile.     

新疆不同地区牛源大肠杆菌耐药性分析

This study was aimed to investigate the resistance of Escherichia coli isolates from cattle farms in Xinjiang to antibiotics. The resistances of Escherichia coli isolates from cattle farms in five regions to commonly used antibiotic were determined using microdilution broth assay. Escherichia coli isolates were highly resistant to ampicillin (up to 39.3%) in all regions, and existed multidrug-resistant, but zero resistance was the dominant. Escherichia coli isolates from E cattle farm in Cherqi town, Baicheng county, exhibited resistant to three kinds of quinolones (6.7%). The results suggested that the resistances of Escherichia coli isolates to commonly used antibiotics were not serious in Xinjiang.     

Occurrence and characterization of verocytotoxigenic Escherichia coli (VTEC) strains from dairy farms in Trinidad

A cross-sectional study was conducted to determine the prevalence and characteristics of verocytotoxigenic Escherichia coli (VTEC) on 25 dairy farms each located in Waller field and Carlsen field farming areas in Trinidad. On each selected farm, faecal samples were collected from milking cows, calves and humans; rectal swabs were obtained from pet farm dogs; bulk milk was sampled as well as effluent from the milking parlour. Escherichia coli was isolated from all sources on selective media using standard methods. Isolates of E. coli were subjected to slide agglutination test using E. coli O157 antiserum, vero cell cytotoxicity assay to detect verocytotoxin (VT) and heat labile toxin (LT) production, the polymerase chain reaction (PCR) to detect VT genes, and the dry spot test to screen for E. coli O157 and non-O157 strains. In addition, faecal samples from animal and human sources were tested for VT genes using PCR. Of a total of 933 E. coli isolates tested by the slide test, eight (0.9%) were positive for the O157 strain. The vero cell cytotoxicity assay detected VT-producing strains of E. coli in 16.6%, 14.6%, 3.2% and 7.1% of isolates from cows, calves, farm dogs and humans respectively (P < 0.05; chi(2)). For LT production, the highest frequency was detected amongst isolates of E. coli from calves (10.8%) and the lowest (0.0%) amongst isolates from humans and bulk milk (P < 0.05; chi(2)). Of the 61 VT-producing isolates by vero cell cytotoxicity assay tested by PCR, the VT, LT and eae genes were detected in 62.3%, 4.9% and 1.6% respectively (P < 0.05; chi(2)). Amongst the 45 E. coli isolates that were VT positive (vero cell) or VT-gene positive by PCR, 2.2%, 2.2%, 4.4% and 6.7% belonged to non-O157 strains O91, O111, O103 and O157, respectively, as determined by the Dry spot test. Detection of VTEC strains in milk and dairy animals poses a health risk to consumers of milk originating from these farms. In addition, the demonstration of VTEC strains in humans, VT gene in faecal samples and E. coli isolates as well as non-O157 VTEC strains of E. coli are being documented for the first time in the country.     

Prevalence of extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella pneumoniae in food-producing animals

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.     

Experimental reproduction of airsacculitis and septicemia by aerosol exposure of 1-day-old chicks using Congo red-positive Escherichia coli

To further demonstrate the association of phenotype and virulence of Congo red Escherichia coli (CREC), experiments were designed to reproduce airsacculitis and colisepticemia in 1-day-old chicks via aerosol exposure. In eight separate experiments in which a total of 462 chicks were exposed to CREC, the mortality rate was 4.11% and the morbidity rate was 13.4%, with both the dead and diseased chicks showing lesions of fibrinous airsacculitis, pericarditis, and perihepatitis. In contrast, when the corresponding non-CREC derivatives (the same E. coli strains, but not expressing the CR phenotype) were used as the aerosol inoculum in five additional experiments using 284 chicks, no chicks died and no lesions were observed. These results clearly show a strong correlation between the expression of the CR phenotype and virulence in avian E. coli.     

Virulence factors of avian Escherichia coli

A total of 45 strains of Escherichia coli isolates from chickens with colisepticemia were examined for virulence factors commonly found in pathogenic groups of E. coli. These strains were studied for the following: pathogenicity in 1-day-old chicks; toxin, hemolysin, and colicin production; cell invasiveness and adherence; hemagglutination for fimbriae detection; serum resistance; aerobactin production in iron-limited conditions; and plasmid content. The characteristics exhibited by virulent strains were invasion for HeLa and chicken fibroblast cells, serum resistance, colicin V, and aerobactin production. None of the isolates were toxigenic or positive in hemagglutination tests. The molecular genetic studies of the virulence factors by agarose electrophoresis showed that the plasmids of these strains are of high molecular weight.