TITLE:
Differentiation of Avian Pathogenic Escherichia coli Strains from Broiler Chickens by Multiplex Polymerase Chain Reaction (PCR) and Random Amplified Polymorphic (RAPD) DNA
AUTHORS:
Dirgam Ahmad Roussan, Hana Zakaria, Ghassan Khawaldeh, Ibrahim Shaheen
KEYWORDS:
Avian Pathogenic Escherichia coli, Broiler, Multiplex PCR, Random Amplification of Polymorphic DNA
JOURNAL NAME:
Open Journal of Veterinary Medicine,
Vol.4 No.10,
October
20,
2014
ABSTRACT: We examined 50 Escherichia coli (E.
coli) strains isolated from broiler chickens between January 2013 to March
2014 in order to evaluate the epidemiological
prevalence of avian pathogenic E. coli (APEC) in Jordan by multiplex PCR
and random amplification of polymorphic DNA (RAPD) tests. The multiplex
polymerase chain reaction (PCR) which was used as tentative criteria of APEC
targets 8 virulence associated genes; enteroaggregative toxin (astA),
Type 1 fimbria adhesion (fimH), iron-repressible protein (irp2),
P fimbriae (papC), aerobactin (iucD), temperature-sensitive hemagglutinin
(tsh), vacuolating autotransporter toxin (vat), and colicin V
plasmid operon (cva/cvi) genes. The number of detected genes
could be used as a reliable index of their virulence. E. coli strains
already typed as an APEC always harbor 5 to 8 genes, but non-APEC strains
harbor less than 4 genes. Assuming the criteria of an APEC is possession of 5
or more virulence associated genes; we found that all 50 E. coli strains
were classified as APEC strains. The RAPD analysis showed that the E. coli strains could be grouped into 35 of RAPD types by using these two different
RAPD primer sets, RAPD analysis primer 4 5'AAGAGCCCGT5', and RAPD analysis
primer 6 5'CCCGTCAGCA3'. The current study confirmed the endemic nature of APEC
in broiler flocks in Jordan. It is essential that the biosecurity on poultry
farms should be improved to prevent the introduction and dissemination of APEC
and other agents. Furthermore, farmers need to be educated about the signs, lesions,
and the importance of this agent.