Paper Information

Journal:   IRANIAN JOURNAL OF VETERINARY RESEARCH (IJVR)   2007 , Volume 8 , Number 4 (21); Page(s) 319 To 324.
 
Paper: 

PREVALENCE OF ESCHERICHIA COLI O157:H7 ON DAIRY FARMS IN SHIRAZ, IRAN BY IMMUNOMAGNETIC SEPARATION AND MULTIPLEX PCR

 
 
Author(s):  SAMI M.*, FIROUZI R., SHEKARFOROUSH SEYED SHAHRAM
 
* DEPARTMENT OF FOOD HYGIENE AND PUBLIC HEALTH, SCHOOL OF VETERINARY MEDICINE, SHAHID BAHONAR UNIVERSITY OF KERMAN, KERMAN, IRAN
 
Abstract: 

To identify the reservoirs of shiga toxin-producing Escherichia coli 0157, sensitive detection and isolation methods are necessary. The sensitivity of traditional culture methods can be improved significantly by the inclusion of an immunoconcentration step, resulting in less false-negative results. In this study, enrichment procedure and immunomagnetic separation (IMS) were compared for use in conjunction with a multiplex polymerase chain reaction (M-PCR) method for detection of genes stx1, stx2, eaeA and hlyA. A total number of 975 faecal samples were collected iTom26 dairy farms in Shiraz area, Shiraz, southern Iran. The samples were cultured at 370C for 18-24 hrs in modified tryptic soy broth (m-TSB). Each of five enriched samples were pooled and examined in two ways-direct PCR and IMS. The detection limit of the M-PCR protocol for seeded E. coli 0157:H7: ATCC: 43895inm-SB withouts tool was 1.23x 102 CFU/ml, whereas it was 1.23 x 106 CFU/ml with enriched faecal sample. In direct PCR of enriched samples, no positive sample was detected. However, in IMS of enriched samples one specimen was positive. The prevalence of E. coli O157:H7 in faeces of cows in examined farms was 0.51% and the herd prevalence was 3.86%. Isolation of this serotype trom faecal sample indicates that cattle are reservoirs of this pathogen and potentially a source of human infection. This finding is of considerable public health importance.

 
Keyword(s): ESCHERICHIA COLI O157:H7, CATTLE, M-PCR, IMS, SHIRAZ
 
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