Isolation and identification of Escherichia coli O157:H7 from ground beef hamburgers in Khuzestan Province, Iran (original) (raw)
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Detection of E. coli O157:H7 in Meat Using Polymerase Chain Reaction Method and Culture Method
International Journal of Basic Science in Medicine
Introduction: Escherichia coli O157:H7, as a pathogenic agent, can be transmitted through the foods including meat, meat products, dairy products, vegetables and water. The World Health Organization has recommended that all countries in the world, especially developing countries, should consider the investigation of E. coli O157:H7 as a research priority. The aim of this study was to determine the frequency of E. coli O157:H7 in meat of cow, sheep, goat, and camel in Kerman province of Iran using culture and polymerase chain reaction (PCR) methods. Methods: In this study, 280 meat samples consisting of sheep (90 specimens), cow (80 specimens), goat (60 specimens) and camel (50 specimens) meats were randomly separated from carcasses from April to July 2018. After the sampling, microbial culture was performed on the samples. Then, suspected E. coli O157:H7 colonies were evaluated by PCR assay. Results: Out of the 280 samples, 73 samples (26%) were contaminated with E. coli. based on b...
Detection of E.COLI O157:H7 in Meat and Its Products by Conventional and Real Time PCR Techniques
Journal of Food and Dairy Sciences
A total of 68 samples including (raw minced meat, Luncheon, Pasterma, and sandwiches of cooked beef, cooked liver, cooked sausage, kofta and burger) were randomly collected from butchers, take away meals' markets and street vendors in different regions of Great Cairo (Cairo, Giza and Shobra El-Khema). The evaluation of microbial load of the collected samples revealed that, no samples contained Total Bacterial Count (TBC) more than the permissible limits, 26.5%, 10.3%, 10.3%, 5.9%, 35.4% and 86.8% of the examined samples contained Total Fungal Count (TFC), Total Coliform Count (TCC), E.coli (Faecal coliform count), Staphylococcus count, Salmonella and Bacillus cereus counts (BC) more than the permissible limits, respectively. The E.coli (Faecal coliform) positive samples (7) together with another 7 samples contained the highest cfu/g of TCC were selected to be tested with Standard (St) as well as Real Time (RT) PCR techniques to detect the presence of E.coli serotype O157:H7. The result of St Polymerase Chain Reaction (PCR) assay indicated positive results (specific bands) in 78.6% of the examined samples. On the other hand, by using specific RT PCR Kit for the detection and differentiation between E.coli O157:H7 and other E.coli serotypes, the obtained results revealed that, all St PCR positive samples showed a separate plateau differed from that obtained from the E.coli O157:H7 positive control sample, indicating that the present serotype (E.coli O55:H7 as clarified by the kit user manual) has nearly the same primer sequence as E.coli O157:H7 which led to false positive result using St PCR technique. This result indicated that, RT PCR is considered as an important, specific and accurate method for the detection and identification of food poisoning organisms.
PubMed, 2012
Enterohemorrhagic Escherichia coli (EHEC) of the O157:H7 serotype is a worldwide zoonotic pathogen responsible for the majority of severe cases of human EHEC disease. The aim of the present study was to investigate the prevalence of E. coli O157: H7/NM in raw meat samples from two provinces of Iran. During a period from March 2010 to March 2011. Two hundred and ninety five raw meat samples were collected from beef (n= 85), camel, (n= 50), sheep (n= 62), goat (n= 60), and water buffalo (n=38). Fourteen (4.7%) of the 295 samples were positive for E. coli O157. The highest prevalence of E. coli O157 was found in beef samples (8.2%), followed by water buffalo (5.3%), sheep (4.8%), camel (2.0%), and goat (1.7%). Of fourteen E. coli O157 isolates, only one was determined to be serotype O157: H7 while 13 were determined as serotype O157: NM. Of the 14 E. coli O157:H7/NM isolates, one, four, two, and one strains were positive for stx1, stx2, eaeA and ehlyA genes, respectively. The prevalence of this organism varied between seasons with the highest prevalence of E. coli O157 occurring in summer (9.3%). The results of this study showed that beef and water buffalo meat are a significant source for human EHEC E. coli O157:H7/NM infection in Iran. The data reported in this study provides some useful baseline in formation for future research such as molecular or epidemiologic works.
Pakistan Journal of Nutrition, 2005
A seven month period from the beginning October to the end of April 2004, a total of 126 ground beef samples were analyzed to determine the incidence of Escherichia coli (E. coli) O157:H7. Among the sampling months, the incidence of EHEC serotypes were only observed in April. Of the 126 ground beef samples, only one ground beef sample was positive for E. coli O157:H7, having a prevalence of 0.79 %. Five samples were found positive for E. coli O157 serotype, having a prevalence of 3.96 %. The results of this study reveal that the occurrence of EHEC serotypes in ground beef in the Kars (Eastern Turkey) seem to be seasonal and the level of E. coli O157 was significantly higher as compared to other studies in Turkey, but it is similar to studies performed in many countries around the world. Of the tested eighteen antibiotics, resistance towards three or more antibiotics were observed among the all isolates.
Meat Science, 2010
The present study was conducted to investigate the presence of Escherichia coli O157 and O157:H7 strains and to detect the presence of the stx1, stx2, and eaeA genes in isolates derived from 200 samples (100 samples from fresh ground beef and 100 samples from raw meatball). The samples were purchased from the Samsun Province in Turkey, over a period of 1 year. Enrichment-based immunomagnetic separation and multiplex polymerase chain reaction were applied for these analyses. E. coli O157 was detected in five of the 200 (2.5%) samples tested (one isolated from ground beef and four from meatball samples), whereas E. coli O157: H7 was not detected in any sample. During the analysis, eight strains of E. coli O157 were obtained. The genes stx1, stx2, and eaeA were detected in two E. coli O157 isolates obtained from two meatball samples, whereas only the eaeA and the stx2 genes were detected in four E. coli O157 strains that were isolated from one meatball sample. None of the stx1, stx2, and eaeA was detected in the E. coli O157 isolates obtained from the ground beef and the one meatball samples.
Detection of Escherichia coli O157:H7 in the beef marketed in Malaysia
Applied and Environmental Microbiology
Twelve strains of Escherichia coli O157:H7 were isolated from 9 of 25 beef samples purchased from retail stores in Malaysia. These strains produced Shiga toxin 2 with or without Shiga toxin 1 and had the eae gene and a 60-MDa plasmid. The antibiograms and the profiles of the arbitrarily primed PCR of the strains were diverse, suggesting that the strains may have originated from diverse sources.
Improvement of the Detection of E. coli O157 in Ground Beef
Escherichia coli 0157:H7 is now recognized as an important human enteropathogen. PCR technology has proven to be very efficient for the detection of E. coli 0157:H7 in incriminated samples. Development of an extraction procedure resulting in 57% ± 6 recovery of E. coli 0157:H7 from seeded ground beef using 0.01 M phosphate buffered saline pH 6.0 with the aid of differential centrifugation. The optimized PCR reaction mixture consisted of 5 µl of 10X PCR mix (final concentrations: 50 mM KCl ,0.01% gelatin, 10 mM Tris-HCl at pH 9.0), 1 mM MgCl , , 1.6 µM each of the two primers, 0.2 mM of dNTPs and Taq polymerase (2.5 units for SLT-1 and 2 5 units for SLT-2) in a final volume of 50 µl. Prior to optimization, the minimum limit of detection for SLT-1 DNA sequence was 200 DNA targets compared to 100 after complete optimization. With SLT-2 DNA sequence, the minimum level of detection prior to optimization was 150 DNA targets compared to 20 after complete optimization. Optimization of the PCR allowed the detection of 10 SLT-1 and 5 SLT-2 targets per one gram of ground beef with the aid of pellet paint and pre-enrichment at 37°C for 5.5 hours.
Determination of Escherichia coli O157:H7 in Chicken Meats Sold in Sanliurfa Region
2016
Escherichia coli O157:H7 has been an important problem of public health in most countries of the world since 1982. This study was therefore aimed to investigate the presence of E. coli O157:H7 in chicken meat samples collected from various markets in Sanliurfa region which was located in Southern Turkey. For this purpose, 155 chicken meat samples were analyzed between September 2005 and February 2006. The samples were plated onto Cefixime Tellurite Supplement and Sorbitol Mac Conkey Agar after enrichment process. Suspected colonies were then analyzed for identification of E. coli O157:H7 as given in materials and methods section. E. coli O157 and E. coli O157:H7 were found in 9 (5.81%) and 3 (1.94%) of the total of 155 samples, respectively. The results showed that control measures should be developed to prevent contamination with this pathogen in chicken meats in this region. To our knowledge, this is the first report of isolation of E. coli O157:H7 from chicken meat samples in Sou...
Meat science, 2010
The present study was conducted to investigate the presence of Escherichia coli O157 and O157:H7 strains and to detect the presence of the stx1, stx2, and eaeA genes in isolates derived from 200 samples (100 samples from fresh ground beef and 100 samples from raw meatball). The samples were purchased from the Samsun Province in Turkey, over a period of 1 year. Enrichment-based immunomagnetic separation and multiplex polymerase chain reaction were applied for these analyses. E. coli O157 was detected in five of the 200 (2.5%) samples tested (one isolated from ground beef and four from meatball samples), whereas E. coli O157: H7 was not detected in any sample. During the analysis, eight strains of E. coli O157 were obtained. The genes stx1, stx2, and eaeA were detected in two E. coli O157 isolates obtained from two meatball samples, whereas only the eaeA and the stx2 genes were detected in four E. coli O157 strains that were isolated from one meatball sample. None of the stx1, stx2, and eaeA was detected in the E. coli O157 isolates obtained from the ground beef and the one meatball samples.
Turkish Journal of Veterinary & Animal Sciences, 2012
This study aimed to determine the prevalence of Escherichia coli O157 in retail red meat and meat products with the Vitek Immunodiagnostic Assay System, including H7 Escherichia coli phage technology (VIDAS ECPT), and a real-time polymerase chain reaction system (LightCycler PCR; LCPCR). A total of 106 red meat and meat product samples were analyzed with VIDAS ECPT and LCPCR. VIDAS ECPT presumptive positive samples were subjected to VIDAS Immuno Concentration E. coli O157, followed by culture and serology. Among the 27 out of 72 (37.50%) red meat samples and 3 out of 34 (8.82%) red meat products that tested positive by VIDAS ECPT, 5.55% and 0.00% were confirmed positive for E. coli O157 (but not H7), respectively. Red meat and red meat product samples were 73.61% and 20.58% positive by LCPCR. The 5.55% prevalence of E. coli O157 in red meats poses a significant risk for consumers and indicates insufficient hygiene management both at the farm and during the slaughtering and meat hand...