A B S T R A C THealthy lambs are one of the major reservoirs of Shiga toxin-producing Escherichia coli (STEC) and it is known as the cause of foodborne diseases (FBD). The work objective is to characterize (STEC) isolates obtained from rectal swabs of healthy lambs herds, a total of 183 samples were obtained from sheep production units of the State of Mexico. E. coli isolates were confirmed through the amplification of the uid A gene. antimicrobial sensitivity pattern was determined through Kirby-Bauer (CLSI, 2012) test and the presence stx 1 , stx 2 and eae genes from isolates by multiplex PCR. Serotyping was performed using specific anti-O and anti-H sera (SERUNAM, Mexico) for 185 Somatic and 56 flagellar antigens. 126 isolates biochemically and molecularly identified as E. coli were obtained, of which 80 did not express any virulence factor and 46 expressed at least some (STEC) virulence factor. The highest percentage of E. coli resistance was for tetracycline 48.7% (39/80), followed by nalidixic acid 13.7% (11/80), gentamicin 6.2% (5/80) and Ciprofloxacin 3.7% (3/80). Resistance to amikacin, cefotaxime and ceftazidime were not detected. A frequency of 46 STEC isolates (36.2%) were obtained, of which 28/46 (22.0%) expressed stx 1 , stx 2 3/46 (6.5%), stx 1 , stx 2 13/46 (10.2%) and eae 2/46 (1.6%). Thirty different serotypes were obtained. The three serotypes with the highest number of isolates (four each) were: O76:H19, O118:H27 and O146:H21 which have been identified as a cause of diarrhea in human population. An isolate of serogroup O104 was obtained, with a significant importance for European public health. In virtue of the discovered serotypes and the virulence factors distribution, we can affirm that the obtained isolates from lambs in the State of Mexico are classifiable as atypical STEC of low virulence.
Sheep represent one of the main reservoirs of diarrheagenic Escherichia coli; this microorganism is an etiological agent of food-borne diseases, therefore, this work aimed to identify and characterize the principal pathotypes of diarrheagenic E. coli obtained through rectal swabs and samples from sheep carcasses slaughtered in an abattoir at the central region of Mexico. The isolates were subjected to bacteriological identification, serotyping; phylogenetic classification; detection for virulence factors, and antimicrobial sensibility. A total of 90 E. coli isolates were obtained, diarrheagenic serotypes with health public relevance were found: O76:H19 (5), O146:H21 (3), O91:H10 (2), O6:NM (1), and O8:NM (1). According to pathotype, 47.7% of total isolates were Shiga toxin-producing E. coli, while 3.3% were enteropathogenic, 2.2% enterotoxigenic, and 1.1% enteroinvasive E. coli; the remaining isolates did not express the genes used to assign them to some pathotype. Regarding the Shiga toxin subtypes, 31/43 (72.09%) were cataloged as stx1c, 11/43 (25.5%), stx1a- stx1c and 1/43 (2.3%) stx1a- stx1d; while for stx2 it was possible identify stx2g 4/7(57.14%), stx2b 1/7 (14.7%) and stx2b-stx2g 2/7 (28.5%). Almost all pathotypes (91–100%) belonged to phylogroup B1. Furthermore, it was observed that the 90 isolates showed an antimicrobial resistance of 100% to nitrofurantoin, followed by ampicillin, tetracycline, and trimethoprim-sulfamethoxazole. These results highlight the importance of diarrheagenic E. coli as a potential risk for public health during the slaughtering process.
Sheep represent one of the main reservoirs of diarrheagenic Escherichia coli; this microorganism is an etiological agent of food-borne diseases; therefore, this work aimed to identify and characterize the principal pathotypes of diarrheagenic E. coli (DEC) obtained through rectal swabs and carcasses samples from sheep slaughtered in an abattoir at the central region of Mexico. The isolates were subjected to bacteriological identification, serotyping; phylogenetic classification; detection for virulence factors, and antimicrobial sensibility. A total of 90 E. coli isolates were obtained. It was observed through 49 E. coli isolates (54%), 8 of them from carcasses, and 43 from feces was DEC. DEC serotypes with health public relevance were found: O76:H19 (n = 5), O146:H21 (n = 3), O91:H10 (n = 1), O6:NM (n = 1), and O8:NM (n = 1). Regarding the presence of Shiga toxin-producing E.coli (STEC), 43/90 (47.7%) isolates have the stx1 w/o stx2 genes, and therefore were assigned as STEC non-O157; only one isolate expressed stx1 and eae genes and was classified as t-STEC (typical STEC). Additionally, 3/90 (3.3%) harbored only the eae gene and were classified as enteropathogenic E. coli (EPEC), the stp gene was found in 2/90 isolates (2.2%) and were classified as enterotoxigenic E. coli (ETEC); 1/90 (1.1%) isolates harboring the ipaH were classified as enteroinvasive E. coli EIEC. Regarding stx1 genes subtypes, stx1c only was found in 60.5% (26/43), followed by stx1a-stx1c 20.9% (9/43) and stx1a-stx1d 2.3% (1/43). The presence of both, stx1 and stx2 genes was found in 7/43 isolates (16.3%) from rectal swabs; the combination stx1c-stx2g was detected in 3/43 isolates (6.9%), while 4 (9.4%) isolates showed different patterns (stx1a-stx1c-stx2g; stx1c-stx2b-stx2g; stx1c-stx2b and stx1a-stx1c-stx2b-stx2g). STEC isolates showed the major diversity of phylogenetic groups, although phylogroup B1 was predominant in 90.6% (39/43) while there was only one isolate (2.3%) in each remaining phylogroup (A, B2, C, and F). All EPEC, ETEC, and EIEC isolates were clustered in phylogroup B1. We observed that 27.9% (12/43) of STEC isolates carried at least one antibiotic resistance: nine isolates expressed the tetB gene, one isolate the tetA gene, two isolates the sul2 gene, one isolate the sul1 and one isolate the sul1-tetB genes. These results highlight the importance of diarrheagenic E. coli as a potential risk for public health during the slaughtering process.
Introduction: Commensal Escherichia coli is defined as bacteria without known virulence factors that could be playing a specific role in some diseases; however, they could be responsible to disseminate antimicrobial resistance genes to other microorganisms. This study aimed to characterize the commensal E. coli isolates obtained from slaughtered sheep in the central region of Mexico. Methodology: Isolates were classified as commensal E. coli when distinctive genes related to diarrheagenic pathotypes (stx1, stx2, eae, bfp, LT, stp, ipaH, and aggR) were discarded by PCR. Identification of serotype, phylogenetic group, and antimicrobial resistance was also performed. Results: A total of 41 isolates were characterized. The phylogenetic groups found were B1 in 37 isolates (90.2%), A in 2 (4.8%), and 1 isolate (2.4%) for C and D groups. Serotypes associated with diarrhea in humans (O104:H2 and O154:NM) and hemolytic uremic syndrome (O8:NM) were detected. Thirty-three isolates (80%) were resistant to ceftazidime, 23 (56%), to tetracycline 8 (19.5%) to ampicillin, and 1 to amikacin. Six isolates (14.6%) were multidrug-resistant. Conclusions: This study provides new information about commensal E. coli in slaughtered sheep, high percentages of resistance to antibiotics, and different profiles of antimicrobial resistance were found, their dissemination constitute a risk factor towards the consuming population.
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