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Front Microbiol. 2016 Jul 4;7:1020. doi: 10.3389/fmicb.2016.01020. eCollection 2016.

Surveillance of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Dairy Cattle Farms in the Nile Delta, Egypt.

Author information

1
Alere Technologies GmbHJena, Germany; InfectoGnostics Research CampusJena, Germany.
2
Department of Animal Hygiene and Zoonoses, Faculty of Veterinary Medicine, Mansoura University Mansoura, Egypt.
3
Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-InstitutJena, Germany; Department of Poultry Disease, Faculty of Veterinary Medicine, Kafrelsheikh UniversityKafr El-Sheikh, Egypt.
4
Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut Jena, Germany.
5
Alere Technologies GmbHJena, Germany; InfectoGnostics Research CampusJena, Germany; Institute for Medical Microbiology and Hygiene, Technical University of DresdenDresden, Germany.

Abstract

INTRODUCTION:

Industrial livestock farming is a possible source of multi-resistant Gram-negative bacteria, including producers of extended spectrum beta-lactamases (ESBLs) conferring resistance to 3rd generation cephalosporins. Limited information is currently available on the situation of ESBL producers in livestock farming outside of Western Europe. A surveillance study was conducted from January to May in 2014 in four dairy cattle farms in different areas of the Nile delta, Egypt.

MATERIALS AND METHODS:

In total, 266 samples were collected from 4 dairy farms including rectal swabs from clinically healthy cattle (n = 210), and environmental samples from the stalls (n = 56). After 24 h pre-enrichment in buffered peptone water, all samples were screened for 3rd generation cephalosporin-resistant Escherichia coli using Brilliance™ ESBL agar. Suspected colonies of putatively ESBL-producing E. coli were sub-cultured and subsequently genotypically and phenotypically characterized. Susceptibility testing using the VITEK-2 system was performed. All suspect isolates were genotypically analyzed using two DNA-microarray based assays: CarbDetect AS-1 and E. coli PanType AS-2 kit (ALERE). These tests allow detection of a multitude of genes and their alleles associated with resistance toward carbapenems, cephalosporins, and other frequently used antibiotics. Serotypes were determined using the E. coli SeroGenotyping AS-1 kit (ALERE).

RESULTS:

Out of 266 samples tested, 114 (42.8%) ESBL-producing E. coli were geno- and phenotypically identified. 113 of 114 phenotypically 3rd generation cephalosporin-resistant isolates harbored at least one of the ESBL resistance genes covered by the applied assays [blaCTX-M15 (n = 105), blaCTX-M9 (n = 1), blaTEM (n = 90), blaSHV (n = 1)]. Alarmingly, the carbapenemase genes blaOXA-48 (n = 5) and blaOXA-181 (n = 1) were found in isolates that also were phenotypically resistant to imipenem and meropenem. Using the array-based serogenotyping method, 66 of the 118 isolates (55%) could be genotypically assigned to O-types.

CONCLUSION:

This study is considered to be a first report of the high prevalence of ESBL-producing E. coli in dairy farms in Egypt. ESBL-producing E. coli isolates with different underlying resistance mechanisms are common in investigated dairy cattle farms in Egypt. The global rise of ESBL- and carbapenemase-producing Gram-negative bacteria is a big concern, and demands intensified surveillance.

KEYWORDS:

CRE; ESBL; Egypt; Escherichia coli; carbapenemases; dairy cattle; genotype; microarray

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