Characterization of shiga toxin-producing Escherichia coli strains isolated from calves in Poland

Fecal samples from 67 3–5-months-old calves with diarrhea were screened for the presence of shiga toxin-producing Escherichia coli (STEC). Several accessory virulence factors genes were also tested. Among 192 E.coli isolates tested, 15 (7.6%) were found to harbour the shiga toxin 1 or 2 (stx1 or stx2) genes. The stx2-carrying samples were further subtyped by PCR for the stx2c, stx2d, and stx2e toxin variants. It was shown that stx2-positive bacteria mainly possessed the stx2c shiga toxin type gene. The enterohemolysin (hlyA) and intimin (eae) genes were found in seven (46.7%) STEC strains whereas the cytotoxic necrotizin factor 1 and 2 or the P fimbrial genes were detected in two isolates only. This study confirmed that calves are a reservoir of STEC strains (with all pathogenicity genes) that may be virulent for humans.     

Prevalence and antimicrobial resistance of porcine O157 and non-O157 Shiga toxin-producing Escherichia coli from India

The aims of this study were to determine the prevalence of Shiga toxin-producing Escherichia coli (STEC) strains in pigs as a possible STEC reservoir in India as well as to characterize the STEC strains and to determine the antimicrobial resistance pattern of the strains. A total of 782 E. coli isolates from clinically healthy (n?=?473) and diarrhoeic piglets (309) belonging to major pig-producing states of India were screened by the polymerase chain reaction (PCR) assay for the presence of virulence genes characteristic for STEC, that is, Shiga toxin-producing gene(s) (stx1, stx2), intimin (eae), enterohemolysin (hlyA) and STEC autoagglutinating adhesin (Saa). Overall STEC were detected in 113 (14.4 %) piglets, and the prevalence of E. coli O157 and non-O157 STEC were 4 (0.5 %) and 109 (13.9 %), respectively. None of the O157 STEC isolates carried gene encoding for H7 antigen (fliCh7). The various combinations of virulence genes present in the strains studied were stx1 in 4.6 %, stx1 in combination with stx2 gene in 5.1 %, stx1 in combination with stx2 and ehxA in 0.6 %, stx1 in combination with stx2 and eae in 0.2 % and stx2 alone in 3.7 %. All STEC isolates were found negative for STEC autoagglutinating adhesin (Saa). The number of STEC isolates which showed resistance to antimicrobials such as ampicillin, tetracycline, streptomycin, lincomycin, nalidixic acid, sulfadiazine, penicillin, gentamicin, kanamycin and ceftriaxone were 100, 99, 98, 97, 95, 94, 92, 88, 85 and 85, respectively. Ninety-seven isolates showed resistance to more than 2 antimicrobials, and 8 resistance groups (R1 to R8) were observed. This study demonstrates that pigs in India harbour both O157 and non-O157 STEC, and this may pose serious public health problems in future.     

Detection of Shiga Toxin‐Producing Escherichia coli and Enteropathogenic Escherichia coli in Faecal Samples of Healthy Mithun (Bos frontalis) by Multiplex Polymerase Chain Reaction

Faecal samples obtained from 190 healthy mithuns were examined for the presence of Escherichia coli. Total one‐hundred and five E. coli isolates were obtained from these samples, which belonged to 55 different serogroups. These isolates were subjected to multiplex polymerase chain reaction (m‐PCR) for detection of stx1, stx2, eaeA and hlyA genes. Twenty‐three (21.90%) E. coli isolates belonging to 14 serogroups revealed the presence of at least one virulence gene when examined by m‐PCR. Nineteen percent and 2.85% of the mithuns were found to carry Shiga toxin‐producing E. coli (STEC) and enteropathogenic E. coli, respectively. stx1 and stx2 genes were found to be prevalent in 7 (6.67%) and 18 (17.14%) of the isolates respectively, whereas eaeA and hlyA genes were found to be carried by three (2.85% each) isolates. Interestingly, none of the STEC isolates belonged to serogroup O157.     

A National Surveillance of Shiga Toxin‐Producing Escherichia coli in Food‐Producing Animals in Japan

To assess the public health risk, the prevalence and anti‐microbial resistance of Shiga toxin‐producing Escherichia coli (STEC) among food‐producing animals were studied throughout Japan. Faecal samples were collected from healthy animals of 272 cattle, 179 pigs, and 158 broilers on 596 farms in all 47 Japanese prefectures. STEC were isolated from 62 (23%) cattle and 32 (14%) pig samples but from no chicken samples. Of the bovine isolates, 19 belonged to serotypes frequently implicated in human disease (O157:H7/non‐motile (NM)/H not typeable, O26:NM/H11/H21/H not typeable, O113:H21, and O145:NM). The eae genes were observed in 37% of bovine isolates; among them one O145:NM and all four O157 isolates possessed eae‐γ1, and one O145:NM, one O103:H11, and all five O26 isolates possessed eae‐β1 gene. Among the swine isolates, stx2e were dominant, and serotypes frequently implicated in human diseases or eae‐positive isolates were not observed. Bovine isolates showed less anti‐microbial resistance, but six isolates of 26:NM/H11 and O145:NM were multi‐resistant and may need careful monitoring. Swine isolates showed various resistance patterns; chloramphenicol resistance patterns were more common than in bovine isolates. This first national study of STEC in the Japanese veterinary field should aid our understanding of Japan's STEC status.     

Incidence of Shiga toxin-producing Escherichia coli in diarrheic calves and its susceptibility profile to antimicrobials and Eugenia uniflora L

The aim of this study was to evaluate the occurrence of Shiga toxin (stx)-producing Escherichia coli (STEC) in diarrheic newborn calves, as well as the resistance profile of this microorganism against antimicrobials routinely used in veterinary therapy. The antimicrobial profile of Eugenia uniflora against E. coli clinical isolates was also analyzed. Specimens from the recto-anal junction mucosa were investigated by using chromogenic medium and identification of E. coli was done using microbiological methods (Gram staining, indole test, methyl red test, Voges-Proskauer test, citrate test, urease test, and hydrogen sulfide test). The stx1 and stx2 genes corresponding to the STEC pathotype were evaluated by using polymerase chain reaction and electrophoresis. The susceptibility profile to antimicrobial agents commonly used in veterinary therapeutic practice and the antimicrobial effect of lyophilized hydroalcoholic extract of E. uniflora L. leaves against E. coli clinical isolates were evaluated by disk diffusion and microdilution methods. Shiga toxin-positive E. coli was identified in 45% of diarrheic newborn calves (stx1 = 23.2%, stx2 = 4.0%, stx1 + stx2 = 18.2%). The frequency of stx-positive E. coli in the bacterial population was equal to 17.0% (168/990 clinical isolates): 97 (9.8%) stx1-positive E. coli, 12 (1.2%) stx2-positive E. coli, and 59 (6.0%) stx1 + stx2-positive E. coli isolates. All stx-positive E. coli analyzed showed resistance to multiple drugs, that is, from 4 to 10 antimicrobials per clinical isolate (streptomycin, tetracycline, cephalothin, ampicillin, sulfamethoxazole + trimethoprim, nitrofurantoin and nalidixic acid, ciprofloxacin, gentamicin, and chloramphenicol). Effective management measures should be implemented, including clinical and laboratory monitoring, in order to promote animal and worker health and welfare, prevent and control the spread of diseases, and ensure effective treatment of infectious diseases. The E. uniflora L. leaves showed inhibition of microbial growth based on the diameter of halos, ranging from 7.9 to 8.0 mm and 9.9 to 10.1 mm for concentrations of 50 and 150 mg/mL, respectively. This plant displayed bacteriostatic action and a minimum inhibitory concentration of 12.5 mg/mL for all clinical isolates. Its clinical or synergistic effects with antimicrobial agents must be determined from clinical and preclinical trials.     

Characterization of non-O157 shiga toxin-producing Escherichia coli isolates from healthy fat-tailed sheep in southeastern of Iran

The objectives of this study were to determine the presence and prevalence of non-O157 shiga toxin-producing Escherichia coli (STEC) isolates from faeces of healthy fat-tailed sheep and detection of phylogenetic background and antibiotic resistance profile of isolates. One hundred ninety-two E. coli isolates were recovered from obtained rectal swabs and were confirmed by biochemical tests. Antibiotic resistance profiles of isolates were detected and phylogenetic background of isolates was determined according to the presence of the chuA, yjaA and TspE4.C2 genetic markers. The isolates were examined to determine stx ₁ , stx ₂ and eae genes. Non-O157 STEC isolates were identified by using O157 specific antiserum. Forty-three isolates (22.40 %) were positive for one of the stx ₁ , stx ₂ and eae genes, whereas 10.42 % were positive for stx ₁ , 19.38 % for eae and 2.60 % for stx ₂ gene. None of the positive isolates belonged to O157 serogroup. Twenty isolates possessed stx ₁ were distributed in A (six isolates), B1 (13) and D (one) phylogroups, whereas stx ₂ positive isolates fell into A (three isolates) and B1 (two) phylogenetic groups. Eighteen isolates contained eae gene belonged to A (five isolates), B1 (seven) and D (six) phylogroups. The maximum and minimum resistance rates were recorded against to penicillin and co-trimoxazole respectively. The positive isolates for stx ₁ , stx ₂ and eae genes showed several antibiotic resistance patterns, whereas belonged to A, B1 and D phylogroups. In conclusion, faeces of healthy sheep could be considered as the important sources of non-O157 STEC and also multidrug-resistant E. coli isolates.     

Antimicrobial resistance in Escherichia coli isolates from frugivorous (Eidolon helvum) and insectivorous (Nycteris hispida) bats in Southeast Nigeria,with detection of CTX-M-15 producing isolates

Thirty-five Escherichia coli isolates obtained from the liver, spleen and intestines of 180 frugivorous and insectivorous bats were investigated for antimicrobial resistance phenotypes/genotypes, prevalence of Extended-Spectrum beta-lactamase (ESBL) production, virulence gene detection and molecular typing. Eight (22.9 %) of the isolates were multidrug resistant (MDR). Two isolates were cefotaxime-resistant, ESBL-producers and harbored the bla_CTX-M-15 gene; they belonged to ST10184-D and ST2178-B1 lineages. tet(A) gene was detected in all tetracycline-resistant isolates while int1 (n = 8) and bla_TEM (n = 7) genes were also found. Thirty-three of the E. coli isolates were assigned to seven phylogenetic groups, with B1 (45.7 %) being predominant. Three isolates were enteropathogenic E. coli (EPEC) pathovars, containing the eae gene (with the variants gamma and iota), and lacking stx1/stx2 genes. Bats in Nigeria are possible reservoirs of potentially pathogenic MDR E. coli isolates which may be important in the ecology of antimicrobial resistance at the human-livestock-wildlife-environment interfaces. The study reinforces the importance of including wildlife in national antimicrobial resistance monitoring programmes.     

An investigation of shedding and super‐shedding of Shiga toxigenic Escherichia coli O157 and E. coli O26 in cattle presented for slaughter in the Republic of Ireland

Shiga toxigenic Escherichia coli (STEC) are an important group of pathogens and can be transmitted to humans from direct or indirect contact with cattle faeces. This study investigated the shedding of E. coli O157 and O26 in cattle at the time of slaughter and factors associated with super‐shedding (SS) animals. Rectoanal mucosal swab (RAMS) samples were collected from cattle (n = 1,317) at three large Irish commercial beef abattoirs over an 18 month period, and metadata were collected at the time of sampling regarding farm of origin, animal age, breed and gender. RAMS swabs were examined for the presence and numbers of E. coli O157 and O26 using a previously developed quantitative real‐time PCR protocol. Samples positive by PCR were culturally examined and isolates analysed for the presence of stx subtypes, eae and phylogroup. Any samples with counts >10⁴ CFU/swab of STEC O157 or O26 were deemed to be super‐shedders. Overall, 4.18% (55/1,317) of RAMS samples were positive for STEC O157, and 2.13% (28/1,317) were classified as STEC O157 SS. For STEC O26, 0.76% (10/1,317) of cattle were positive for STEC O26, and 0.23% (3/1,317) were classified as super‐shedders. Fewer STEC shedders and SS were noted among older animals (>37 months). There was a seasonal trend observed for STEC O157, with the highest prevalence of shedding and SS events in the autumn (August to October). The majority of E. coli O157 (50/55) isolates had stx2 and were eae positive, with no significant difference between SS and low shedders (LS). Interestingly, all STEC O26 (n = 10) were eae negative and had varied stx profiles. This study demonstrates that, while the overall shedding rates are relatively low in cattle at slaughter, among positive animals there is a high level of SS, which may pose a higher risk of cross‐contamination during slaughter.     

Virulence factors in Escherichia coli isolated from calves with diarrhea in Vietnam

This study was conducted to determine the prevalence and characteristics of pathogenic Escherichia (E.) coli strains from diarrheic calves in Vietnam. A total of 345 E. coli isolates obtained from 322 diarrheic calves were subjected to PCR and multiplex PCR for detection of the f5, f41, f17, eae, sta, lt, stx1, and stx2 genes. Of the 345 isolates, 108 (31.3%) carried at least one fimbrial gene. Of these 108 isolates, 50 carried genes for Shiga toxin and one possessed genes for both enterotoxin and Shiga toxin. The eae gene was found in 34 isolates (9.8%), 23 of which also carried stx genes. The Shiga toxin genes were detected in 177 isolates (51.3%) and the number of strains that carried stx1, stx2 and stx1/stx2 were 46, 73 and 58, respectively. Among 177 Shiga toxin-producing E. coli isolates, 89 carried the ehxA gene and 87 possessed the saa gene. Further characterization of the stx subtypes showed that among 104 stx1-positive isolates, 58 were the stx1c variant and 46 were the stx1 variant. Of the 131 stx2-positive strains, 48 were stx2, 48 were stx2c, 11 were stx2d, 17 were stx2g, and seven were stx2c/stx2g subtypes. The serogroups most prevalent among the 345 isolates were O15, O20, O103 and O157.     

Development of a PCR‐Based Method for Specific Identification of Genotypic Markers of Shiga Toxin‐Producing Escherichia coli Strains

A simple, rapid and specific PCR‐based method for identification of shiga toxin‐producing Escherichia coli (STEC) was developed. The procedure involves amplification of the E. coli‐specific universal stress protein A (uspA) gene (uspa‐PCR), with the primer pair described by other authors, which allows differentiation of E. coli (STEC and non‐STEC) from other gram‐negative bacteria followed by identification of the main genetic virulence traits of the uspA‐positive isolates. For this purpose, two multiplex PCR assays, based on previously published primer sequences, were established. Assay 1 (mPCR‐1) uses three primer pairs and detects the genes encoding O157 (rfb), enterohemolysin (ehly) and shiga toxin (stx), generating amplification products of 420, 534 and 230 bp, respectively. Assay 2 (mPCR‐2) uses four primer pairs specific for rfb (E. coli O157), eaeA (intimin), stx1 and stx2 (shiga toxin 1 and 2, respectively), generating PCR amplicons of 420, 840, 348 and 584 bp, respectively. These two assays were validated by testing several E. coli reference strains and 202 previously characterized E. coli isolates originating from calves and from children, and 100% agreement with previous results was obtained. The method developed can be used for specific identification of STEC bacteria including those of the O157 serogroup.     

Diversity of Shiga toxin-producing Escherichia coli in sheep flocks of Paraná State,southern Brazil

Sheep constitute an important source of zoonotic pathogens as Shiga toxin-producing Escherichia coli (STEC). In this study, the prevalence, serotypes and virulence profiles of STEC were investigated among 130 healthy sheep from small and medium farms in southern Brazil. STEC was isolated from 65 (50%) of the tested animals and detected in all flocks. A total of 70 STEC isolates were characterized, and belonged to 23 different O:H serotypes, many of which associated with human disease, including hemolytic-uremic syndrome (HUS). Among the serotypes identified, O76:H19 and O65:H– were the most common, and O75:H14 and O169:H7 have not been previously reported in STEC strains. Most of the STEC isolates harbored only stx1, whereas the Stx2b subtype was the most common among those carrying stx2. Enterohemolysin (ehxA) and intimin (eae) genes were detected in 61 (87.1%) and four (5.7%) isolates, respectively. Genes encoding putative adhesins (saa, iha, lpf_O113) and toxins (subAB and cdtV) were also observed. The majority of the isolates displayed virulence features related to pathogenesis of STEC, such as adherence to epithelial cells, high cytotoxicity and enterohemolytic activity. Ovine STEC isolates belonged mostly to phylogenetic group B1. PFGE revealed particular clones distributed in some farms, as well as variations in the degree of genetic similarity within serotypes examined. In conclusion, STEC are widely distributed in southern Brazilian sheep, and belonged mainly to serotypes that are not commonly reported in other regions, such as O76:H19 and O65:H–. A geographical variation in the distribution of STEC serotypes seems to occur in sheep.     

Occurrence of genes encoding aminoglycoside-modifying enzymes in Escherichia coli isolates from chicken meat

ABSTRACT

1. The aim of the experiment was to determine the occurrence of genes encoding aminoglycoside-modifying enzymes (AMEs) in Escherichia coli isolates recovered from chicken meat.

2. Antibiotic sensitivity was tested using the disc diffusion test. AMEs and virulence profile were determined by PCR/sequencing.

3. Out of 195 meat samples collected, 185 (95%) isolates were identified as E. coli. Disc diffusion showed a resistance value of 22% (n = 42) for at least one of the antibiotic aminoglycosides (AGs) tested (tobramycin, gentamycin, amikacin and kanamycin). PCR screening showed the presence of three classes of AMEs, namely, aac(3)-II (12%), aac(6?)-Ib (7%) and aac(2?)-Ia (5%). Eight of the 42 isolates were positive for the stx₁ and sxt₂ genes and were defined as Shiga toxin-producing E coli., while the eae gene was positive in one strain. Among the 42 isolates, group A was the predominant phylogenetic identified (76%), followed by group D (21%). One isolate belonged to subgroup B2₃.

4. The results suggested that chicken meat could be an important reservoir of AMEs, and pose a potential risk by dissemination of resistance to humans through the food chain.     

Non‐O157 Shiga Toxin‐producing Escherichia coli Isolated from Diarrhoeic Calves in Argentina

Faecal samples from 76 diarrhoeic calves belonging to 36 farms located in the Pampas plain, Argentina, were examined for Shiga toxin‐producing Escherichia coli (STEC). A total of 15 STEC strains were isolated from 12 (15.8%) calves which came from six different farms. All stx positive strains assayed by PCR were also positives in the Vero cell cytotoxicity test. The majority (60.0%) of the STEC strains carried the stx₁ gene. Twelve (80.0%) of the STEC isolates which belonged to serotypes O5:H‐ (n = 4), O26:H11 (n = 4), O26:H‐ (n = 1), O111:H‐ (n = 2), and O123:H38 (n = 1) were also enterohaemolysin (EHly) positive and carried the gene encoding for intimin (eae). All the stx positive strains were negative for the bfpA gene. Localized adherence to HEp‐2 cells were observed in 83.3% of the eae+ STEC strains. STEC belonging to serotype O5:H‐ showed atypical biochemical properties, including urease production. Urease was also produced by two strains belonging to serotypes O153:H? and non‐typeable, respectively. Resistance to three or more antibiotics was observed in 12 (80.0%) of the STEC isolates. Most of the serotypes of STEC recovered in this survey carried virulence traits that are associated with increased human and bovine pathogenicity. The present study shows that highly virulent STEC strains are being shed by diarrhoeic calves from farms located in a high incidence area of human STEC infections.     

High level of multidrug-resistant Escherichia coli in young dairy calves in southern Vietnam

This study investigated the occurrence of antimicrobial-resistant Escherichia coli in dairy calves in southern Vietnam. Fecal samples were taken directly from the rectum of 84 calves from 41 smallholder dairy farms, when newborn and at 14 days of age for isolation of E. coli. Escherichia coli strains were isolated from 144 of the 168 fecal samples tested. Of the 144 E. coli isolates, 40% were found to be susceptible to all 12 antimicrobial drugs tested and 53% of the E. coli isolates were resistant to at least three antimicrobials. Calves were colonized with antimicrobial-resistant E. coli already on the day of birth. Resistance to tetracycline was most common, followed by resistance to sulfamethoxazole, ampicillin, trimethoprim, and ciprofloxacin. Four isolates carried a gene encoding for extended-spectrum cephalosporinases (ESC), and these genes belonged to bla_CTX-M group 1 (2 isolates), bla_CTX-M group 9 (1 isolate), and bla_CMY-2 (1 isolate). Thirty-three isolates had a plasmid-mediated quinolone resistance (PMQR) phenotype, and 30 of these carried the qnrS gene. These results are of importance for management routines of dairy cattle to prevent the spread of antimicrobial resistance.

     

Phenotypic and genotypic properties of Escherichia coli isolated from colisepticemic cases of Japanese quail

This study was conducted to characterize the Escherichia coli isolates from colisepticemic Japanese quails. One hundred and nine E. coli were isolated in pure culture from heart blood of dead Japanese quails. The sampled birds were originated from four different farms. Antibiotic resistance pattern of E. coli isolates were determined against nine antibacterial agents. Phylotype and virulence genes of the isolates were detected by polymerase chain reaction. By disc diffusion method, all of the isolates showed resistance to three or more antibiotics, and 19 different patterns of multiple drug resistance were observed. Phylotyping of the most prevalent multiple drug-resistant isolates revealed that they mostly belonged to phylogroups A (A₁ subgroup). The E. coli isolates belong to four phylogenetic groups: A (55.0%), B1 (18.3%), B2 (17.4%), and D (9.2%). Eighty-nine (81.7%) isolates were distributed in five phylogenetic subgroups including 22 (20.2%) in A₀, 38 (34.9%) in A₁, 19 (17.4%) in B2₃, 7 (6.4%) in D₁, and 3 (2.8%) in D₂. The examined E. coli isolates exhibit at least one of the virulence genes tested, whereas three most prevalent genes were crl (94.5%), fimH (89.0%), and iutA (51.4%), respectively. The genetic marker for Afa (afaI B-C), S (sfa/focD-E), and P (papE-F) fimbriae were found in one, four, and ten isolates, respectively. Thirteen different combinations of virulence gene were observed, where combination of crl and fimH genes was the most prevalent pattern. None of the isolates contained the ipaH, stx1, stx2, and eaeA genetic markers. In conclusion, E. coli strains could be considered as a causative agent of mortality in quail farms. In conclusion, E. coli isolates from colisepticemic quails are distributed in different phylogroups, are resistant to combinations of antibiotic agents, and contain several virulence genes.     

Shiga toxin-producing Escherichia coli O157:H7 from extensive cattle of the fighting bulls breed

Shiga toxin-producing Escherichia coli (STEC) O157:H7 represents a major public health concern worldwide, with cattle recognized as their main natural reservoir. The aim of this work was to determine the prevalence and the pheno-genotypic characteristics of STEC O157:H7 in a herd with 268 cattle of the fighting bulls breed (De Lidia breed) managed under extensive conditions in the South-West of Spain. Rectal-anal swabs of all animals were collected and examined for STEC O157:H7 by performing an immunomagnetic concentration and separation procedure combined with PCR, and the resulting isolates were characterized by both phenotypic and genotypic methods. Overall, STEC O157:H7 was isolated from seven animals (2.6%) in the herd. The PCR procedure indicated that all seven isolates displayed stx₂, eae-γ1, ehxA, O157 rfbE, and fliCh7 genes. They belonged to phage types 4 (one isolate) and 42 (two isolates), and four isolates reacted with typing phages but did not conform to a recognized pattern. Among the seven isolates there were five indistinguishable PFGE patterns and other two which differed only in ?2 restriction fragments, supporting the existence of horizontal transmission among animals in the herd. The present study demonstrates that cattle managed under extensive conditions in Spain can excrete STEC O157:H7 with their faeces. To our knowledge this is the first isolation of this pathogen from De Lidia cattle.     

Pathotyping and antimicrobial susceptibility testing of Escherichia coli isolates from neonatal calves

Neonatal calf mortality is a major concern to livestock sector worldwide. Neonatal calf diarrhoea (NCD), an acute severe condition causes morbidity and mortality in calves. Amongst various pathogens involved in NCD, E. coli is considered as one of the major causes. The study was targeted to characterize E. coli isolates from neonatal calves for diarrhoeagenic Escherichia coli (DEC) types (pathotyping), antimicrobial resistance (AMR) profiling and to correlate with epidemiological parameters. From neonates, a total of 113 faecal samples were collected, out of that 308, lactose fermenting colonies were confirmed as E. coli. Pathotypable isolates (12.3%) were represented by STEC (6.1%), EPEC (2.9%), ETEC (1.9%), EAEC (0.9%) and EHEC (0.3%). Occurrence of STEC was more in non-diarrhoeic calves, whereas ETEC was observed more in diarrhoeic calves. EPEC occurrence was observed in both diarrhoeic and non-diarrhoeic calves. Fishers extract test showed no significant association for occurrence of DEC types to type of dairies, health status, species, breed, age and sex of neonatal calves. Two hundred and eighty isolates were tested for antimicrobial susceptibility. The isolates showed maximum resistance towards ampicillin (55.4%) followed by tetracycline (54.3%), while minimum resistance was observed towards meropenem (2.5%). Multidrug resistant E. coli isolates were found to be 139 (49.6%), and Extended-spectrum beta-lactamase (ESBL) producers were 120 (42.9%). DEC pathotypes like STEC, ETEC, EHEC and EAEC that are also multidrug resistant present in neonatal calves have zoonotic potential and hence are of public health significance.

     

Detection and characterization of Shiga toxin-producing Escherichia coli in feral pigeons

Escherichia coli strains producing a variant of Shiga toxin 2 (Stx2), designated Stx2f, have been recently described in the stools of feral pigeons. During 1997-1998, 649 pigeons were trapped and examined in three different squares of Rome. Stool samples were collected from each bird and enrichment cultures were examined for the presence of Stx by the vero cell assay. Stx-producing E. coli (STEC) were isolated from the positive cultures and characterized by serotyping and PCR analysis of stx and other virulence-related genes. Stx was detected in 10.8% of the stool enrichment cultures. The percentage of positive birds did not differ significantly for the three flocks considered and the season of sample collection. Conversely, STEC carriage was significantly more frequent in young than in adult birds (17.9 versus 8.2%). None of the birds examined showed signs of disease. STEC strains were isolated from 30 of 42 Stx-positive cultures examined. All the strains produced Stx2f, and most of them possessed genes encoding for intimin and the cytolethal distending toxin (CLDT). Six serogroups were identified, but most of the isolates belonged to O45, O18ab, and O75. Molecular typing indicated that most of the isolates within a flock were clonally-related. This work confirms that pigeons represent a natural reservoir of STEC strains characterized by the production of the toxin variant Stx2f, and by the frequent presence of eae and cldt genes. Further work is needed to clarify whether these STEC may represent a cause of avian disease or even a potential health hazard for humans.     

Subtype analysis of stx1, stx2 and eae genes in Shiga toxin-producing Escherichia coli (STEC) and typical and atypical enteropathogenic E. coli (EPEC) from lambs in India

Seventy-five Escherichia coli isolates with at least one targeted virulence gene were recovered from 338 lambs with (n=230) and without (n=108) diarrhoea. The isolates belonged to 36 different serogroups. Shiga toxin-producing E. coli (STEC) was isolated from 9.6% of lambs with and 24.1% of lambs without diarrhoea. Enteropathogenic E. coli (EPEC) was isolated from 6.1% of lambs with and 11.1% of lambs without diarrhoea. Of 26 EPEC isolates, seven were typical (positive for bfpA), and, of 34 stx(1) positive isolates, 25 were subtyped as stx(1c). Five of 29 stx(2) positive isolates were subtyped as stx(2d) and two as stx(2c). Seven of 45 eae positive isolates were subtyped as eae subtype zeta (eaezeta). This appears to be the first report of the isolation of typical EPEC from sheep in India.