scholarly journals Isolation of Shiga toxin-producing strains of Escherichia coli from beef and swine carcasses and the characterization of their genes

2018 ◽  
Vol 9 (2) ◽  
pp. 275-780 ◽  
Author(s):  
O. М. Berhilevych ◽  
V. V. Kasianchuk ◽  
O. M. Deriabin ◽  
M. D. Kukhtyn

Escherichia coli is part of the normal microflora of the intestinal tract of humans and warm-blooded animals, but its presence in raw material and food of animal origin is considered as fecal contamination and can be very dangerous for consumers. The determination of the number of E. coli in raw material and food is important because among them can be pathogenic strains. The most dangerous strains are considered enterohemorrhagic E. coli as a causative agent of severe bloody diarrhea and hemorrhagic uremic syndrome in humans through the production of Shiga-toxin, which is the main virulence factor, responsible for disease. The aim of this study was to identify the prevalence of Shiga toxin-producing strains of E. coli (STEC) from swabs of beef and swine carcass in slaughterhouses in Ukraine and characterize their genes, which are responsible for pathogenic properties. A total of 230 samples of swabs from beef (130) and swine (100) carcasses were obtained from 5 slaughterhouses in Ukraine between 2012 and 2015. Samples of swabs from carcasses were randomly selected at the final point of the process after the final washing of the carcass from the following areas: distal hind limb, abdomen (lateral and medial) from swine carcasses, brisket, flank and flank groin areas from beef carcasses. All samples were examined by culture-dependent method, after that each positive isolate of STEC was analyzed by multiplex PCR to detect the stx1, stx2, and eae genes. Out of 230 collected samples, seven (7.2%) were contaminated with STEC. The highest prevalence of STEC was found in swabs from beef carcasses (8.1%) in comparison to swabs from swine carcasses (5.7%). The stx1 gene was the predominant gene detected in all STEC positive samples. The eae gene was found in one of the examined isolates from beef carcass. Three isolates from swabs of beef carcass carried both stx1 and stx2 genes, one isolate showed association between stx1 and eae genes, one isolate was positive for stx1 gene only. In swabs from swine carcasses (2 isolates) stx1 and stx2 genes were presented simultaneously. The results of this study suggested that fresh raw meat could be a potential vehicle for transmission of the Shiga toxin-producing strain of E. coli to humans. This is the first report of STEC prevalence in beef and swine carcasses in Ukraine and these data will be valuable for microbiological risk assessment and help the appropriate services to develop strategies to mitigate health risk.

2020 ◽  
Vol 152 ◽  
pp. 15667-15675
Author(s):  
Chakirath Folakè Arikè Salifou ◽  
Cyrille Boko ◽  
Isidore Houaga ◽  
Raoul Agossa ◽  
Isabelle Ogbankotan ◽  
...  

Objectives: The study aimed to search for E. coli O157 and non-O157 in milk, meat and faeces of cattle, sheep and pigs slaughtered in Cotonou. Methodology and Results: One hundred and Seventy-Five (175) samples including 25 meat, 25 faeces per species and 25 milk from cattle were analysed for E. coli O157; O26 and O111 and the virulence genes were identified by PCR. The SAS software (1998) and the bilateral Z test were used to calculate and compare the identification frequencies. E. coli O157 was identified in 4% of cattle faeces, 4% of sheep faeces, and 20% of beef and, in 20% of milk samples. E. coli O26 was identified in 12% of cattle faeces and, in 8% of beef samples. E. coli O111 was identified at frequencies of 8%, and 12% in faeces of sheep and pigs, respectively. The eae gene was detected in 4% of beef, ovine meat, milk, pig faeces and in sheep faeces. stx1 was detected in 8% of milk, and in 4% of bovine and sheep faeces. The strains possessing the gene were all of E. coli O157 with the exception of one from pig faeces identified as O111. Conclusions and application of findings: The presence of these serogroups of E. coli with virulence genes poses a real food safety problem in Benin. This study findings must be taken into account for risk assessment and management related to the consumption of food of animal origin. Keywords: Benin, E. coli O157, O26, O111, faeces, meat, milk


2021 ◽  
Author(s):  
Million Weldeselassie ◽  
Getachew Gugsa ◽  
Yisehak Tsegaye ◽  
Nesibu Awol ◽  
Ashwani Kumar ◽  
...  

Abstract Background: Escherichia coli O157:H7 is an emerging and major zoonotic foodborne pathogen with an increasing concern for the emergence and spread of antimicrobial-resistant strains which may results in sporadic cases to serious outbreaks in the whole world. Cattle have been identified as a major reservoir of the pathogen. This study aimed to isolate and characterize Shiga toxin-producing E. coli O157:H7 from raw milk, yogurt, and meat of bovine origin and determine their antimicrobial susceptibility pattern. Methods: A cross-sectional study was conducted from December 2014-June 2015 and a total of 284 milk and meat were collected from different sources in Mekelle. The collected samples were analyzed for the presence of E. coli and Shiga toxin-producing E. coli O157:H7 and determination of their antimicrobial susceptibility pattern following the standard bacteriological and molecular techniques and procedures, and antimicrobial sensitivity test. Results: Out of the total 284 samples, 70(24.64%) were positive bacteriologically to E. coli and 14.29% were found to be Shiga toxin-producing E. coli O157:H7. All (100%) E. coli isolates carried the pal and 41.67% eae gene (EHEC). Of these EHEC isolates 40% and 60% were positive for stx1 and stx2, respectively. E. coli isolates were showed the highest level of sensitivity for Gentamycin (91.7%) but the highest level of resistance to Amoxicillin (95.8%). Of the tested isolates, 18(75%) of E. coli showed multiple antimicrobial resistance.Conclusions: The current study revealed the occurrence of Shiga toxin-producing E. coli O157:H7 in foods of bovine origin in the study area. So, there is a chance of acquiring infection via the consumption of raw or undercooked food of bovine origin. Thus, awareness creation should be made on foodborne disease caused by Shiga toxin-producing E. coli O157:H7 with due consideration on the safe handling and consumption of food of animal origin.


2016 ◽  
Vol 14 (1) ◽  
pp. 63-68 ◽  
Author(s):  
MM Akter ◽  
S Majumder ◽  
KH MNH Nazir ◽  
M Rahman

Shiga toxin-producing Escherichia coli (STEC) are zoonotically important pathogen which causes hemorrhagic colitis, diarrhea, and hemolytic uremic syndrome in animals and humans. The present study was designed to isolate and identify the STEC from fecal samples of diarrheic cattle. A total of 35 diarrheic fecal samples were collected from Bangladesh Agricultural University (BAU) Veterinary Teaching Hospital. The samples were primarily examined for the detection of E. coli by cultural, morphological and biochemical characteristics, followed by confirmation of the isolates by Polymerase Chain Reaction (PCR) using gene specific primers. Later, the STEC were identified among the isolated E. coli through detection of Stx-1 and Stx-2 genes using duplex PCR. Out of 35 samples, 25 (71.43%) isolates were confirmed to be associated with E. coli, of which only 7 (28%) isolates were shiga toxin producers, and all of them were positive for Stx-1. However, no Stx-2 positive isolate could be detected. From this study, it may be concluded that cattle can act as a reservoir of STEC which may transmit to human or other animals.J. Bangladesh Agril. Univ. 14(1): 63-68, June 2016


2004 ◽  
Vol 133 (2) ◽  
pp. 359-366 ◽  
Author(s):  
M. ČOBELJIĆ ◽  
B. DIMIĆ ◽  
D. OPAČIĆ ◽  
Z. LEPšANOVIĆ ◽  
V. STOJANOVIĆ ◽  
...  

Faecal samples of 2660 domestic animals from 116 farms and 956 samples of food were examined for the presence of Shiga toxin-producing Escherichia coli (STEC). STEC was recovered from 126 (15·3%) cattle, 135 (11·3%) pigs, 135 (66·8%) sheep, 31 (73·8%) goats, 4 (1%) chicken, and 15 (1·6%) food samples. Of all STEC isolates, 21·5, 25·8 and 15% produced enterohaemolysin, α-haemolysin, and aerobactin respectively, 1·6% displayed localized adherence (LA) to HEp-2 cells, 27·6% were sorbitol negative, and 30% were resistant to antibiotics. Only 14 (3·1%) of the STEC isolates belonged to human infection-associated serogroups (O26, O55, O111, O128 and O157), designated as enterohaemorrhagic E. coli (EHEC). This study revealed that STEC are prevalent in domestic animals, and to a lesser extent in food of animal origin in Serbia, but the absence of a EHEC phenotypic profile (characteristic serogroup, LA, enterohaemolysin production) in most animal STEC strains may explain the low incidence of human STEC infection in this part of the world.


2015 ◽  
Vol 78 (12) ◽  
pp. 2136-2142 ◽  
Author(s):  
BRITTNEY R. SCOTT ◽  
XIANG YANG ◽  
IFIGENIA GEORNARAS ◽  
ROBERT J. DELMORE ◽  
DALE R. WOERNER ◽  
...  

Studies were conducted to (i) determine whether inoculants of nonpathogenic Escherichia coli biotype I effectively served as surrogates for E. coli O157:H7, non-O157 Shiga toxin–producing E. coli, and Salmonella when prerigor beef carcass tissue was treated with a commercially available blend of lactic acid and citric acid (LCA) at a range of industry conditions of concentration, temperature, and pressure; (ii) determine the antimicrobial efficacy of LCA; and (iii) investigate the use of surrogates to validate a hot water and LCA sequential treatment as a carcass spray intervention in a commercial beef harvest plant. In an initial laboratory study, beef brisket tissue samples were left uninoculated or were inoculated (~6 log CFU/cm2) on the adipose side with E. coli O157:H7 (5-strain mixture), non-O157 Shiga toxin–producing E. coli (12-strain mixture), Salmonella (6-strain mixture), or nonpathogenic E. coli (5-strain mixture). Samples were left untreated (control) or were treated with LCA, in a spray cabinet, at one of eight combinations of solution concentration (1.9 and 2.5%), solution temperature (43 and 60°C), and application pressure (15 and 30 lb/in2). In a second study, the E. coli surrogates were inoculated (~6 log CFU/cm2) on beef carcasses in a commercial facility to validate the use of a hot water treatment (92.2 to 92.8°C, 13 to 15 lb/in2) followed by an LCA treatment (1.9%, 50 to 51.7°C, 13 to 15 lb/in2, 10 s). In the in vitro study, surrogate and pathogen bacteria did not differ in their response to the tested LCA treatments. Treatment with LCA reduced (P < 0.05) inoculated populations by 0.9 to 1.5 log CFU/cm2, irrespective of inoculum type. The hot water and LCA sequential treatments evaluated in the commercial facility reduced (P < 0.05) the inoculated nonpathogenic E. coli surrogates on carcasses by 3.7 log CFU/cm2. This study therefore provides the meat industry with data for this sequential multiple hurdle system for the operation parameters described.


2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Geresu Minda Asfaw ◽  
Regassa Shimelis

Escherichia coli O157 : H7 (E. coli O157 : H7) has been found to be the major cause of food-borne diseases and a serious public health problem in the world, with an increasing concern for the emergence and spread of antimicrobial-resistant strains. Hitherto, little is known about the carriage of E. coli O157 : H7 and its antimicrobial susceptibility profile in the food of animal origin in Ethiopia. This study aimed to determine the occurrence and multidrug resistance profile of E. coli O157 : H7 from food of animal origin at different catering establishments in the selected study settings of Arsi Zone. One hundred ninety-two animal origin food items, namely, raw/minced meat (locally known as “Kitfo,” “Kurt,” and “Dulet”), raw milk, egg sandwich, and cream cake samples were collected and processed for microbiological detection of E. coli O157 : H7. Out of 192 samples, 2.1% (4/192) were positive for E. coli O157 : H7. Two E. coli O157 : H7 isolates were obtained from “Dulet” (6.3%) followed by “Kurt” (3.1%, 1/32) and raw milk (3.1%, 1/32), whereas no isolate was obtained from “Kitfo,” egg sandwich, and cream cake samples. Of the 4 E. coli O157 : H7 isolates subjected to 10 panels of antimicrobial discs, 3 (75%) were highly resistant to kanamycin, streptomycin, and nitrofurantoin. Besides, all the isolates displayed multidrug resistance phenotypes, 3 to 5 antimicrobial resistance, amid kanamycin, streptomycin, nitrofurantoin, tetracycline, and chloramphenicol. The occurrence of multidrug-resistant E. coli O157 : H7 isolates from foods of animal origin sampled from different catering establishments reveals that the general sanitary condition of the catering establishments, utensils used, and personnel hygienic practices did not comply with the recommended standards. Thus, this finding calls for urgent attention toward appropriate controls and good hygienic practices in different catering establishments dealing with consuming raw/undercooked foods of animal origin.


2019 ◽  
Vol 12 (10) ◽  
pp. 1584-1590
Author(s):  
Maria Kristiani Epi Goma ◽  
Alvita Indraswari ◽  
Aris Haryanto ◽  
Dyah Ayu Widiasih

Background and Aim: The feasibility assessment of food products on the market becomes one of the milestones of food safety. The quality of food safety of animal origin especially pork need to get attention and more real action from the parties related and concerned. Since pork is also a source of transmission for the contagion of foodborne disease so that the study of the existence of several agents in the pork and its products become the benchmark of safety level. This study aimed to isolate, identify, and detect the Shiga toxin 2a (stx2a) gene from Escherichia coli O157:H7 in pork, pig feces, and clean water in the Jagalan slaughterhouse. Materials and Methods: A total of 70 samples consisting of 32 pork samples, 32 pig fecal samples, and 6 clean water samples were used to isolate and identify E. coli O157:H7 and the stx2a gene. Isolation and identification of E. coli O157:H7 were performed using culture on eosin methylene blue agar and Sorbitol-MacConkey agar media and confirmed molecularly with polymerase chain reaction to amplify the target genes rfbE (317 bp) and fliC (381 bp). The isolates, which were identified as E. coli O157:H7, were investigated for the stx2a gene (553 bp). Results: The results of this study show that of the total collected samples, E. coli O157:H7 was 28.6% in Jagalan slaughterhouse and consisted of 25% of pork samples, 31.25% of pig fecal samples, and 33.3% of clean water samples. The isolates that were identified to be E. coli O157:H7 mostly contained the stx2a gene, which was equal to 75%, and consisted of seven isolates from pork samples, seven isolates from fecal samples, and one isolate from clean water samples. Conclusion: E. coli O157:H7 was found in 28.6% of pork, pig feces, and clean water in Jagalan slaughterhouse and 75% of identified E. coli O157:H7 contained the stx2a gene.


2018 ◽  
Vol 120 (7) ◽  
pp. 1457-1473 ◽  
Author(s):  
Edwin Barrios-Villa ◽  
Gerardo Cortés-Cortés ◽  
Patricia Lozano Zarain ◽  
Sergio Romero-Romero ◽  
Norarizbeth Lara Flores ◽  
...  

Purpose Broad-spectrum cephalosporin resistance is rapidly increasing in Escherichia coli, representing a food safety problem. The purpose of this paper is to characterize eight extended-spectrum-ß-lactamase (ESBL) and acquired AmpC ß-lactamase-producing E. coli isolates and virotypes associated, obtained from chicken and pork food samples in Puebla, Mexico. Design/methodology/approach Samples (36 from chicken and 10 from pork) were cultured on Levine agar plates supplemented with cefotaxime (2 mg/L) for isolation of cefotaxime-resistant (CTXR) E. coli. CTXR-E. coli isolates were detected in 33 of 46 samples (72 percent), and one isolate/sample was characterized (28 from chicken and 5 from pork), for ESBL production, phylogenetic group, sequence typing, resistance and virulence genes by PCR and sequencing. Findings Results showed 16 ESBL-E. coli (35 percent) (12/16 belonging to phylogroup B1) and 8 CMY-2-E. coli (17 percent). ESBL detected were as follows (number of isolates): CTX-M-2 (8); CTX-M-1 (2); CTX-M-15 (1); SHV-2a (4) and TEM-52c (1). In total, 20 different sequence types (STs) were identified among the ESBL- or CMY-2-producing E. coli strains, which included four new ones. The CTX-M-15 β-lactamase was detected in one E. coli ST617-ST10 Cplx-B1 strain that also carried ibeA gene. One CMY-2-positive strain of lineage ST224-B2 was detected and it carried the qnrA1 gene. Originality/value In this study, a ST131-based virotyping scheme for strains from food of animal origin was established since this kind of strains constitutes an important vehicle of virulent ESBL- and CMY-2-producing E. coli isolates, which could be transmitted to humans by direct contact or through the food chain.


Author(s):  
B Shome ◽  
Rakshit Ojha ◽  
Suresh Mendem ◽  
Devi Murugesan ◽  
Gopalakrishna Sivaraman ◽  
...  

Summary: Antibiotic-resistant bacteria are emerging threat to the global public health. Estimates suggests >10 million deaths by 2050 due to AMR alone. Six Indian institute collaborated along with 4 UK universities to address the issue of antimicrobial resistance (AMR) through One Health approach under NEOSTAR project jointly funded by Department of Biotechnology (DBT), Government of India and UK Research and Innovation (UKRI), United Kingdom. To address a question “Dose AMR in livestock contributes to AMR in people? A pilot study was conducted which is a collaborative effort to collect archival microbial isolates of Escherichia coli, Klebsiella spp., Staphylococcus aureus and coagulase negative S. aureus (CoNS). The current study deals with E. coli isolates collected from ICAR-NIVEDI, ICAR-CIFT, ICAR-IVRI, ICAR-RCNEH, IIT-Delhi and GMCH and were subjected to QC analysis at ICAR-NIVEDI’s BSL2+ facility, these isolates were further segregated into 5 components viz., Animal origin (including aquaculture) (n=50) food of animal origin (n=30), Human (community settings) (n=3), Hospital origin (n=15) and Environment origin (n=14). A total of 112 E. coli isolates sequenced for whole genome through Illumina’s HiSeq 2500 platform at Sangers Institute, UK and paired end libraries were generated for all the 112 isolates were assembled and subjected to genome characterization and gene mining through various bioinformatics tools revealed the frequency of resistance, co-resistance, and resistant genes are high and similar across Human-Animal-Aquaculture-Environment continuum in India. This emphasizes the need to collaborate and mitigate antibiotic resistance with a ‘One Health’ approach.


2019 ◽  
Vol 82 (8) ◽  
pp. 1440-1448 ◽  
Author(s):  
ALESSANDRA BARLAAM ◽  
ANTONIO PARISI ◽  
ELISA SPINELLI ◽  
MARTA CARUSO ◽  
PIETRO DI TARANTO ◽  
...  

ABSTRACT Antimicrobial resistance in bacteria represents one of the most important challenges for public health worldwide. Human infections from antimicrobial-resistant bacteria can be transmitted from person to person, via the environment (especially in the hospital environment), or via handling or eating contaminated foods. Colistin is well known as a last-resort antibiotic for the treatment of human infections; a recent study performed in the People's Republic of China has revealed that colistin resistance is also conferred by the plasmid-mediated mcr-1 gene in Escherichia coli. After that discovery, further plasmid-mediated, colistin resistance genes have been detected. However, to date, only reports on E. coli carrying the mcr-1 gene (E. coli mcr-1+) in foodstuff are available. E. coli mcr-1+ has been isolated from food of animal origin and vegetables; this discovery has opened a debate among food safety experts. This review aims to provide a critical overview of the currently available scientific literature on the presence of the plasmid-mediated, colistin resistance gene E. coli mcr-1 in foodstuffs, focusing on the main implications and future perspectives for food safety.


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