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The mcr-1 gene in 125 ESBL-producing Gram-negative bacteria obtained from animal products was screened and the location and transferability of the mcr-1 gene were examined. The mcr-1 gene was detected by PCR in one Escherichia coli isolate (RC263) obtained from hard cheese made from raw milk. Antimicrobial susceptibility test showed that the strain RC263 was phenotypically multidrug-resistant with colistin and nine other antimicrobials. Further genetic analysis indicated that this isolate also harboured the bla TEM and bla CTX-M-9 genes. S1-PFGE followed by Southern blotting verified that the mcr-1 gene was located on an approximately 153 kb plasmid. The plasmid was transferred to a recipient E. coli strain by conjugation, suggesting that the mcr-1 gene may spread to other Gram-negative bacteria. In conclusion, ESBL-producing E. coli carrying a transferable plasmid with mcr-1 gene was present in hard cheese in Egypt. [ABSTRACT FROM AUTHOR]

Subclinical mastitis (SCM) is regarded as both a problem for dairy producers and a threat to human health worldwide owing to the potential bacterial contamination of milk and dairy products, particularly those made from raw milk. In the present study, we isolated Escherichia coli from 14 (9.3%) SCM milk samples. We serotyped each E. coli isolate (n = 14), and investigated its potential pathogenicity and antimicrobial resistance (AMR). The serotyping results showed that the E. coli isolates belonged to serotypes O55:H7 (n = 2), O111:H4 (n = 2), O127:H6 (n = 2), O128:HUT (n = 2), O26:HUT (n = 1), O44:H18 (n = 1), O114:H21 (n = 1), O86:HUT (n = 1), O124:HUT (n = 1), and O127:H7 (n = 1). Potential pathogenicity was detected in 93% (13/14) of the isolates. In particular, 13 isolates possessed at least one of the examined virulence genes. Ten isolates (71%) exhibited AMR to at least one of the tested antimicrobials, four (40%) were multidrug-resistant, and one isolate produced extended-spectrum β-lactamases. The obtained results indicate that SCM acts as a source for the spread of potentially pathogenic E. coli strains that are resistant to many groups of antimicrobials, and may constitute a hazard to both public and animal health.

The goal of this study was to examine antimicrobial resistance and characterize the implicated genes in 222 isolates of Escherichia coli from 187 samples of raw milk and the two most popular cheeses in Egypt. E. coli isolates were tested for susceptibility to 12 antimicrobials by a disk diffusion method. Among the 222 E. coli isolates, 66 (29.7%) were resistant to one or more antimicrobials, and half of these resistant isolates showed a multidrug resistance phenotype (resistance to at least three different drug classes). The resistance traits were observed to tetracycline (27.5%), ampicillin (18.9%), streptomycin (18.5%), sulfamethoxazole-trimethoprim (11.3%), cefotaxime (4.5%), kanamycin (4.1%), ceftazidime (3.6%), chloramphenicol (2.3%), nalidixic acid (1.8%), and ciprofloxacin (1.4%). No resistance to fosfomycin and imipenem was observed. Tetracycline resistance genes tetA, tetB, and tetD were detected in 53 isolates, 9 isolates, and 1 isolate, respectively, but tetC was not detected. Aminoglycoside resistance genes strA, strB, aadA, and aphA1 were detected in 41, 41, 11, and 9 isolates, respectively. Sulfonamide resistance genes sul1, sul2, and sul3 were detected in 7, 25, and 3 isolates, respectively. Of 42 ampicillin-resistant isolates, bla TEM , bla CTX-M , and bla SHV were detected in 40, 9, and 3 isolates, respectively, and 10 (23.8%) ampicillin-resistant isolates were found to produce extended-spectrum β-lactamase. Each bla gene of extended-spectrum β-lactamase-producing E. coli was further subtyped to be bla CTX-M-15 , bla CTX-M-104 , bla TEM-1 , and bla SHV-12 . The class 1 integron was also detected in 28 resistant isolates, and three different patterns were obtained by PCR-restriction fragment length polymorphism. Sequencing analysis of the variable region revealed that four isolates had dfrA12/orfF/aadA2, two had aadA22, and one had dfrA1/aadA1. These data suggest that antimicrobial-resistant E. coli are widely distributed in the milk production and processing environment in Egypt and may play a role in dissemination of antimicrobial resistance to other pathogenic and commensal bacteria.

The objectives of this study were to investigate prevalence and pathogenic potential of Escherichia coli contaminating raw milk and its products in Egypt. Out of 187 dairy products including 72 raw milk samples, 55 Karish cheese and 60 Ras cheese, 222 E. coli isolates including 111, 89 and 22 were obtained from 55 raw milk samples (76.4%), 41 Karish cheese (74.5%), and 13 Ras cheese (21.7%), respectively. Isolated E. coli strains were examined for 24 representative virulence genes present in diarrheagenic E. coli (DEC) and extraintestinal pathogenic E. coli (ExPEC). Among DEC and ExPEC virulence factors, genes for enteropathogenic E. coli (eaeA, bfpA, EAF), enterohemorrhagic E. coli (stx1, stx2, eaeA), enterotoxigenic E. coli (elt, est), enteroinvasive E. coli (invE), enteroaggregative E. coli (Eagg, astA), diffusely adherent E. coli (daaD), ExPEC (cdt-I to cdt-V, cnf1, cnf2, hlyA) and putative adhesins (efa1, iha, ehaA, saa, and lpfAO113) were screened by colony hybridization assay. Out of 222 E. coli strains, 104 (46.8%) isolated from 69 (36.9%) samples carried one or more virulence genes. The most prevalent gene detected was lpfAO113 (40.5%), followed by ehaA (32.4%,), astA (3.15%,), iha (1.80%), hlyA (1.35%), stx1 (0.90%), stx2 (0.90%), eaeA (0.45%), cdt-III (0.45%) and cnf2 (0.45%). Two strains isolated from Karish cheese harbored 5 virulence genes (stx1, stx2, iha, ehaA, lpfAO113). Stx subtype was determined to be stx1 (not stx1c or stx1d) and stx2d. Indeed, expression of hemolysin A, CDT-III, CNF-II, Stx1 and Stx2d was confirmed by blood agar plate, cytotoxicity assay and Western blotting, respectively. Among the 222 E. coli strains, 54 (48.6%), 38 (42.6%) and 12 (54.7%) isolated from raw milk, Karish cheese and Ras cheese were potentially virulent, respectively. O-genotyping indicated that most of the potentially virulent E. coli isolates did not belong to clinically important O serogroups except O75, O91 and O166, which have been associated with human diseases. Phylogenetic grouping revealed that 150 (67.6%), 67 (30.2%) and 5 (2.30%) strains were clustered into A, B1 and D groups, respectively, which are considered to be associated with intestinal infection, indicating that these E. coli strains might have a potential to cause gastroenteritis. To the best of our knowledge, this is the first comprehensive study regarding prevalence and pathogenic potential of E. coli in dairy products in Egypt. Raw milk, Karish cheese and Ras cheese in Egypt are highly contaminated with E. coli including potentially pathogenic strains, which may impose a public health threat., (Copyright © 2016 Elsevier B.V. All rights reserved.)