Prevalence and molecular characterisation of Shiga toxin-producing Escherichia coli in sheep farms of Sanandaj, Iran

Abstract

Shiga toxin-producing Escherichia coli (STEC) strains have emerged as important foodborne pathogens of global public health concern, causing life-threatening diseases. Animals and their products have been documented as important reservoirs for STECs, especially E. coli O157. The aim of this study was to investigate STECs from healthy and diarrhoeic sheep in Sanandaj, Iran. In the current study, a total of 81 sheep faecal samples were taken (22 from diarrhoeic sheep and 59 from healthy sheep). E. coli and subsequently STEC strains was detected according to standard protocol (cultural characterisation and PCR assays). Finally, the frequency of Shiga-toxin producing gene(s) (stx1, stx2), intimin (eaeA) and enterohaemolysin (hlyA) was detected among STEC isolates using duplex PCR. Totally, 42 E. coli were isolated from 81 faecal samples (51.85% contamination). Of these, 34 isolates (80.9%) were identified as STEC patotypes based on Sorbitol-MacConkey (SMAC) medium culturing and also the presence of stx1 and/or stx2. Of these, only 3 isolates (7.1%) were identified as serotype O157:H7 based on PCR assay. In addition, the results showed that STEC bacteria were significantly more prevalent in diarrhoeic samples than in healthy samples (50% vs. 22.1%). Overall, the PCR results showed that 33 (97%), 12 (35.3%) and 8 (23.5%) isolates carried stx1, stx2 and hlyA, respectively. The eaeA gene was not found in any isolate. The number of isolated STEC bacteria in spring (10 isolates) and winter (14 isolates) were significantly higher than those in summer (4 isolates) and autumn (6 isolates) (P=0.039). Also, the number of STEC in diarrhoea samples was significantly higher compared to non-diarrhoea samples (P=0.032). In conclusion, the present study revealed high prevalence rate of STEC including serotype O157:H7 and non-O157:H7 in sheep faeces which highlights the importance of sheep as a reservoir of STEC pathogen in Sanandaj region. Therefore, additional control and preventive measures must be undertaken to control the contamination by this pathogen.