Journal of Food, Agriculture and Environment

Vol 12, Issue 2,2014
Online ISSN: 1459-0263
Print ISSN: 1459-0255

Characterisation of Escherichia coli isolates recovered from bovine clinical and subclinical mastitic milk


Abdullah A. Al-Arfaj 1, Ashgan M. Hessain 2 *, Mohamed I. AlHazmi 3, Mustafa A. Gassem 3, Moussa I. Mohamed 1

Recieved Date: 2014-02-03, Accepted Date: 2014-04-20


The objective of this study was the molecular characterisation of shiga toxogenic E. coli in clinical and subclinical mastitic milk collected from different Egyptian governorates by multiplex Polymerase Chain Reaction. A total of 292 bulk-tank milk samples were collected from different Egyptian governorates. Bacteriological examination of milk samples revealed the presence of 44 Escherichia coli isolates (15.068%). The result showed that the E. coli incidence in clinical mastitic milk was higher than in the subclinical mastitic milk. Serotyping of the E. coli isolates yielded from bacteriological examination of milk samples revealed 41 strains out of 44 E. coli isolates typed (15.06%). Detection of verotoxin activity of E. coli strains using vero cells investigated showed that 5 strains (6.4%) belonged to O128:K67, O111:K58 and O126:K58 from clinical mastitic milk samples. Meanwhile, 15 strains (7.01%) were recovered from subclinical mastitic cases. For detection of shiga toxin type 2 (stx2) and intimin (eaeA) genes multiplex PCR was used. Positive amplification of intimin gene was found from the DNA extracted from 4 strains of O111 (9.76%), 3 strains of O128:K67 (7.32%), 2 strains of O26:K60, O55:K59 and O119:K69 (4.88%) and 1 strain of O126:K7 (2.44%). Results also revealed positive simultaneous amplification of 255 bp fragment of shiga toxin type 2 gene and 384 bp fragment of intimin gene from all E. coli of serovar O157:H7 13, 22, 3 strains of serovar O111 (7.32%), 2 strains of O128 and O26:K60 (4.88%) and 1 strain of O55:K59 (2.44%), which confirm that multiplex PCR assay is useful for identification of STEC possessing the eaeA and stx2 genes.


Mastitis, E. coli, multiplex PCR, shiga toxin type 2, intimin gene

Journal: Journal of Food, Agriculture and Environment
Year: 2014
Volume: 12
Issue: 2
Category: Food and Health
Pages: 198-201

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