scholarly journals Prevalence, molecular characterization and antimicrobial profiles of Enterohaemorrhagic E. coli O157 isolated from ruminants slaughtered in Al Ain, United Arab Emirates

2020 ◽  
Author(s):  
Dawood Al-Ajmi ◽  
Shafeeq Rahman ◽  
Sharmila Banu
Keyword(s):  
United Arab Emirates ◽  
Shiga Toxin ◽  
Meat Products ◽  
Immunomagnetic Separation ◽  
Latex Agglutination ◽  
Human Infection ◽  
Human Consumption ◽  
Fecal Samples ◽  
Sheep And Goats ◽  
E Coli

Abstract Background: Shiga toxin-producing Escherichia coli (STEC) are associated with major food illness around the world. E.coli O157, has been widely reported as the most common STEC serogroup, and has emerged as an important enteric pathogen. Further, cattle have been identified as a major E. coli O157:H7 reservoir for human infection; however, the ecology of this organism in camels, sheep and goats is less understood. The current study aims to evaluate the prevalence of E. coli serotype O157 in feces of cattle, camels, sheep and goats slaughtered in United Arab Emirates (UAE) for meat consumption. This study was carried out on fecal samples of healthy cattle (n = 137), camels (n = 140), sheep (n = 141) and goats (n = 150) during the period of September 2017 to August 2018. We have used the traditional sensitive immunomagnetic separation technique (IMS) coupled with a culture plating method for detection of E. coli O157. Non-sorbitol fermenting colonies were assessed via the latex agglutination test and the positive cultures were subjected to PCR for detection of attaching and effacing genes (eaeA), hemolysin A (hlyA) and Shiga toxin-producing genes (stx1 and stx2) and genes specific for E. coli O157:H7 (rfb O157, uid A and flic H7). All E. coli O157 isolates were analyzed for their susceptibility pattern toward 20 select antibiotics.Results: E. coli O157 was present in the fecal samples of goats, camels and cattle at 2%, 3.3%, and 1.6%, respectively. In sheep we failed to detect any E. coli O157 strains. The most prevalent E.coli O157 gene identified across all species’ isolates was stx2, while stx1 was not detected in any of the samples. ­After testing samples from camels, goats and cattle, Cefotaxime (100%), Chloramphenicol (100%), Ciprofloxacin (100%), Norfloxacin (100%) and Polymixin B (100%) showed susceptibility showed susceptibility to all E.coli O157 isolates.Conclusion: This is the first study, to our knowledge, to report on the prevalence of E. coli O157 in the slaughter animals in UAE and clearly demonstrates the presence of these pathogens in slaughtered animals, which could possibly contaminate the meat products intended for human consumption.

scholarly journals Occurrence, virulence genes, and antimicrobial profiles of Escherichia coli O157 isolated from ruminants slaughtered in Al Ain, United Arab Emirates

2020 ◽  
Author(s):  
Dawood Al-Ajmi ◽  
Shafeeq Rahman ◽  
Sharmila Banu
Keyword(s):  
Escherichia Coli ◽  
United Arab Emirates ◽  
Shiga Toxin ◽  
Meat Products ◽  
Immunomagnetic Separation ◽  
Latex Agglutination ◽  
Human Consumption ◽  
Fecal Samples ◽  
Chain Reactions ◽  
E Coli

Abstract Background: Shiga toxin-producing Escherichia coli (STEC) is a major source of food-borne illness around the world. E. coli O157 has been widely reported as the most common STEC serogroup and has emerged as an important enteric pathogen. Cattle, in particular have been identified as a major E. coli O157:H7 reservoir of human infections; however, the prevalence of this organism in camels, sheep, and goats is less understood. The aim of this study was to evaluate the occurrence and concentration of E. coli serotype O157 in the feces of healthy camels (n = 140), cattle (n =137), sheep (n = 141) and goats (n = 150) slaughtered in United Arab Emirates (UAE) for meat consumption between September 2017 and August 2018. We used immunomagnetic separation coupled with a culture-plating method to detect E. coli O157. Non-sorbitol fermenting colonies were assessed via latex-agglutination testing, and positive cultures were analyzed by performing polymerase chain reactions to detect genes encoding attaching and effacing protein (eaeA), hemolysin A (hlyA, also known as ehxA) and Shiga toxin (stx1 and stx2), and E. coli O157:H7 specific genes (rfb O157, uidA, and fliC). All E. coli O157 isolates were analyzed for their susceptibility to 20 selected antimicrobials.Results: E. coli O157 was observed in camels, goats, and cattle fecal samples at abundances of 4.3%, 2%, and 1.46%, respectively, but it was undetectable in sheep feces. The most prevalent E. coli O157 gene in all STEC isolates was stx2;, whereas, stx1 was not detected in any of the samples. The fecal samples from camels, goats, and cattle harbored E. coli O157 isolates that were 100% susceptible to cefotaxime, chloramphenicol, ciprofloxacin, norfloxacin, and polymyxin B.Conclusion: To our knowledge, this is the first report on the occurrence of E. coli O157 in slaughter animals in the UAE. Our results clearly demonstrate the presence of E. coli O157 in slaughtered animals, which could possibly contaminate meat products intended for human consumption.

scholarly journals Prevalence, Virulence Genes and Antimicrobial Profiles of Escherichia Coli O157:H7 Isolated from Healthy Cattle

2021 ◽  
Author(s):  
Ghassan Tayh ◽  
Salma Mariem Boubaker ◽  
Rym Ben Khedher ◽  
Mounir Jbeli ◽  
Faten Ben Chehida ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Latex Agglutination ◽  
Human Consumption ◽  
Toxin Gene ◽  
Fecal Samples ◽  
E Coli ◽  
Food Borne ◽  
Healthy Cattle ◽  
Amoxicillin Clavulanic Acid

Abstract Background: Shiga toxin-producing Escherichia coli (STEC) O157:H7 is associated with intestinal infection in human and considered a main cause of food-borne diseases. It was isolated from animals, human and food. The aim of the study was to assess the incidence of E. coli O157:H7 in fecal samples of healthy cattle collected in slaughterhouses (n=160) and from farms (n=100).Methods: E. coli isolates were detected on MacConkey agar. A total of 236 E. coli isolates were recovered from fecal samples of healthy cattle. We used sorbitol MacConkey to detect non-sorbitol fermenting colonies that were examined for the presence of O157 antigen by latex agglutination, and positive bacteria were screened for the existence of stx1, stx2, eaeA and ehxA by PCR as well as rfbEO157 and fliCH7 genes specific for serotype O157. All isolates were examined for the susceptibility against 21antibiotics discs.Results: Of the 236 E. coli isolates, 4.2% (10/236) were positive for STEC O157:H7. Shiga toxin gene (stx2) was present in 70% of isolates, stx1 and ehxA were confirmed in 60% of the isolates, whereas eae was identified in two isolates. Other virulence factors screened (fimH, sfa/focDE, cdt3, traT, iutA and hly) were present among the 10 isolates. All E. coli O157:H7 isolates were sensitive to amoxicillin/clavulanic acid, cefotaxime, cefepime, aztreonam, colistin and sulfamethoxazole/trimethoprim. All isolates belong to the phylo-group E.Conclusion: This is the first study of the incidence of E. coli O157:H7 in cattle in Tunisia. Our finding proves the existence of STEC O157:H7 in healthy animals producing food for human consumption which could be a source of human contamination.

National Survey of Shiga Toxin–Producing Escherichia coli Serotypes O26, O45, O103, O111, O121, O145, and O157 in Australian Beef Cattle Feces

2016 ◽  
Vol 79 (11) ◽  
pp. 1868-1874 ◽  
Author(s):  
GLEN E. MELLOR ◽  
NARELLE FEGAN ◽  
LESLEY L. DUFFY ◽  
KATE E. McMILLAN ◽  
DAVID JORDAN ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Beef Cattle ◽  
Shiga Toxin ◽  
Immunomagnetic Separation ◽  
The United States ◽  
Pcr Analysis ◽  
Fecal Samples ◽  
E Coli ◽  
Beef Products ◽  
Low Prevalence

ABSTRACT Escherichia coli O157 and six non-O157 Shiga toxin–producing E. coli (STEC) serotypes (O26, O45, O103, O111, O121, and O145, colloquially referred to as the “big 6”) have been classified as adulterants of raw nonintact beef products in the United States. While beef cattle are a known reservoir for the prototype STEC serotype, E. coli O157, less is known about the dissemination of non-O157 STEC serotypes in Australian cattle. In the present study, 1,500 fecal samples were collected at slaughter from adult (n =628) and young (n =286) beef cattle, adult (n =128) and young (n =143) dairy cattle, and veal calves (n = 315) across 31 Australian export-registered processing establishments. Fecal samples were enriched and tested for E. coli O157 and the big 6 STEC serotypes using BAX System PCR and immunomagnetic separation methods. Pathogenic STEC (pSTEC; isolates that possess stx, eae, and an O antigen marker for O157 or a big 6 serotype) were isolated from 115 samples (7.7%), of which 100 (6.7%) contained E. coli O157 and 19 (1.3%) contained a big 6 serotype. Four of the 115 samples contained multiple pSTEC serotypes. Among samples confirmed for big 6 pSTEC, 15 (1%) contained E. coli O26 and 4 (0.3%) contained E. coli O111. pSTEC of serotypes O45, O103, O121, and O145 were not isolated from any sample, even though genes indicative of E. coli belonging to these serotypes were detected by PCR. Analysis of animal classes revealed a higher pSTEC prevalence in younger animals, including veal (12.7%), young beef (9.8%), and young dairy (7.0%), than in adult animals, including adult beef (5.1%) and adult dairy (3.9%). This study is the largest of its kind undertaken in Australia. In contrast to E. coli O157 and consistent with previous findings, this study reports a relatively low prevalence of big 6 pSTEC serotypes in Australian cattle populations.

Prevalence of non-O157 Shiga Toxin-producing E. Coli in Children and Calves in Al- Muthanna Province, Iraq

2021 ◽  
Vol 14 (2) ◽  
pp. 78-90
Author(s):  
Ahmed Jarad ◽  
Kh. Al- Jeboori
Keyword(s):  
Shiga Toxin ◽  
Latex Agglutination ◽  
Macconkey Agar ◽  
Biochemical Tests ◽  
Bacteriological Study ◽  
E Coli ◽  
Additional Information ◽  
Single Colony ◽  
Big Six ◽  
Reliable Isolation

The present study focus on non-O157 Shiga toxin-producing E. Coli (STEC), included a bacteriological study was subjected to provide additional information for non-O157 STEC prevalence in children and calves. Isolation by using selective culturing media (CHROMagar STEC and CHROMagar O157) from 127 children suffering from diarrhea and 133 calves in Al- Muthanna province. Characterization depends on culturing positive colony on MacConkey agar and Levin’s Eosin Methylene blue agar, staining single colony from the growth by gram stain, biochemical tests; Indole, the Methyl Red, Voges-Proskauer, Citrate test, Oxidase, Catalase, Urease, Motility, Kligler Iron and Api-20E, were done to confirm a diagnosis of non-O157 STEC, The reliable isolation as non-O157 STEC serotyping by specific latex agglutination test for the target non-O157 STEC (big six) serogroup (O26, O45, O103, O111, O121 and O145). The current study showed the prevalence of non-O157 STEC was 20 of out 127 (15.73%) in samples collected from children and 27 / 133 (20.30%) in calves samples in conclusion the Non-O157 STEC is an important cause of diarrhea in children, and calves; finally, the calves play an important reservoir for Non-O157 STEC.

scholarly journals Prevalence and molecular detection of shiga toxin producing Escherichia coli from diarrheic cattle

2016 ◽  
Vol 14 (1) ◽  
pp. 63-68 ◽  
Author(s):  
MM Akter ◽  
S Majumder ◽  
KH MNH Nazir ◽  
M Rahman
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Molecular Detection ◽  
Chain Reaction ◽  
Specific Primers ◽  
Fecal Samples ◽  
E Coli ◽  
Uremic Syndrome ◽  
Polymerase Chain ◽  
Positive Isolate

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

scholarly journals Comparison between human and animal isolates of Shiga toxin-producing Escherichia coli O157 from Australia

2002 ◽  
Vol 128 (3) ◽  
pp. 357-362 ◽  
Author(s):  
N. FEGAN ◽  
P. DESMARCHELIER
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Human Infection ◽  
Pulsed Field Gel Electrophoresis ◽  
Toxin Gene ◽  
Escherichia Coli O157 ◽  
Animal Population ◽  
E Coli ◽  
Virulence Markers ◽  
Animal Isolates

There is very little human disease associated with enterohaemorrhagic Escherichia coli O157 in Australia even though these organisms are present in the animal population. A group of Australian isolates of E. coli O157:H7 and O157:H- from human and animal sources were tested for the presence of virulence markers and compared by XbaI DNA macrorestriction analysis using pulsed-field gel electrophoresis (PFGE). Each of 102 isolates tested contained the gene eae which encodes the E. coli attaching and effacing factor and all but one carried the enterohaemolysin gene, ehxA, found on the EHEC plasmid. The most common Shiga toxin gene carried was stx2c, either alone (16%) or in combination with stx1 (74%) or stx2 (3%). PFGE grouped the isolates based on H serotype and some clusters were source specific. Australian E. coli O157:H7 and H- isolates from human, animal and meat sources carry all the virulence markers associated with EHEC disease in humans therefore other factors must be responsible for the low rates of human infection in Australia.

Comparison of Rapid Enzyme-Linked Immunosorbent Assay and Immunomagnetic Separation Methods for Detection of Escherichia coli O157 in Fecal, Hide, Carcass, and Ground Beef Samples

2007 ◽  
Vol 70 (10) ◽  
pp. 2230-2234 ◽  
Author(s):  
T. W. THOMPSON ◽  
T. P. STEPHENS ◽  
G. H. LONERAGAN ◽  
M. F. MILLER ◽  
M. M. BRASHEARS
Keyword(s):  
Escherichia Coli ◽  
Ground Beef ◽  
Immunomagnetic Separation ◽  
Separation Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Escherichia Coli O157 ◽  
Perfect Agreement ◽  
Fecal Samples ◽  
E Coli ◽  
Beef Products

Rapid enzyme-linked immunosorbent assays (ELISAs) are approved for detection of Escherichia coli O157 in beef products. However, these kits have also been used in the industry to detect this pathogen on hides or in feces of cattle, although this use has not been validated. The objective of this study was to compare commercially available ELISAs (E. coli Now, Reveal, and VIP) with immunomagnetic separation along with selective media to detect E. coli O157 on hides, in feces, and in medium- and low-level-inoculated ground beef and carcasses (simulated by using briskets) samples. Naturally infected hide and fecal samples were subjected to both the immunomagnetic separation method and ELISAs for the detection of E. coli O157. Additionally, E. coli O157 inoculated and noninoculated ground beef and beef briskets were used to simulate meat and carcass samples. When comparing the detection results from the ELISAs (E. coli Now, Reveal, and VIP) to the immunomagnetic separation method, poor agreement was observed for fecal samples (kappa = 0.10, 0.02, and 0.03 for E. coli Now, Reveal, and VIP, respectively), and fair-to-moderate agreement was observed for hide samples (kappa = 0.30, 0.51, and 0.29 for E. coli Now, Reveal, and VIP, respectively). However, there was near-perfect agreement between the immunomagnetic separation method and ELISAs for ground beef (kappa = 1, 1, and 0.80 for E. coli Now, Reveal, and VIP, respectively) and brisket (kappa = 1, 1, and 1 for E. coli Now, Reveal, and VIP, respectively) samples. Assuming immunomagnetic separation is the best available method, these data suggest that the ELISAs are not useful in detecting E. coli O157 from hide or fecal samples. However, when ELISAs are used on ground beef and beef brisket samples they can be used with a high degree of confidence.

scholarly journals 449 Late-Breaking: Impact of feeding a propriety yeast-based synbiotic product on fecal shedding of top-7 Shiga toxin-producing Escherichia coli in feedlot cattle

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 141-141
Author(s):  
Raghavendra Amachawadi ◽  
Xiaorong Shi ◽  
LeighAnn George ◽  
Miles Theurer ◽  
Twig Marston ◽  
...  
Keyword(s):  
Shiga Toxin ◽  
Virulence Genes ◽  
Multivariable Analysis ◽  
Feedlot Cattle ◽  
Control Group ◽  
Fecal Samples ◽  
Fecal Shedding ◽  
E Coli ◽  
Multiplex Pcr Assay ◽  
The Impact

Abstract Shiga toxin-producing E. coli (STEC) belonging to serogroups O26, O45, O111, O103, O121, O145, and O157, called ‘top-7’, are major foodborne pathogens. Cattle are a major reservoir, in which STEC colonize the hindgut and are shed in the feces, which is a major source of contamination of food. Our objective was to evaluate the impact of a proprietary yeast-based synbiotic product (prebiotic and probiotic; Alltech, Inc., Nicholasville, KY) on fecal shedding of top-7 STEC in feedlot cattle. Twenty existing pens, housing 40–112 steers per pen, with an estimated 60 to 90 days to slaughter, were randomly assigned to a control group or a treatment group that received 22 g of the synbiotic product per steer per day, as a top dress, in a finishing diet. Twenty pen-floor fecal samples were collected from each pen on days 0, 21, 42, and 54. Fecal samples were enriched and subjected to a multiplex PCR assay targeting serogroup-specific genes for the top-7 STEC and three major virulence genes, stx1 (Shiga toxin 1), stx2 (Shiga toxin 2), and eae (intimin). Bivariate descriptive statistics for the major serogroups and virulence genes were assessed prior to multivariable analysis using mixed effects logistic regression. The overall prevalence of the top-7 serogroups were 44.5% of O26, 41.3% of O157, 15.1% of O103, 13.7% of O45, 7.8% of O121, and 0.6% of O111. The overall prevalence of stx1, stx2, and eae were 43.9%, 70.8%, and 49%, respectively. E. coli O26, O157, and O45 had a significant treatment and sampling day interaction (P < 0.0001). On d 42, fecal samples from treated group had lower prevalence (P < 0.01) of O26, O103, and O45 compared to the control group. In conclusion, the in-feed administration of the synbiotic product appears to reduce fecal shedding of certain top-7 STEC serogroups in the feedlot cattle.

Virulence Characterization and Molecular Subtyping of Typical and Atypical Escherichia coli O157:H7 and O157:H(−) Isolated from Fecal Samples and Beef Carcasses in Mexico

2015 ◽  
Vol 78 (2) ◽  
pp. 264-272 ◽  
Author(s):  
CLAUDIA NARVÁEZ-BRAVO ◽  
ALEJANDRO ECHEVERRY ◽  
MARKUS F. MILLER ◽  
ARGENIS RODAS-GONZÁLEZ ◽  
M. TODD BRASHEARS ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Genes ◽  
Immunomagnetic Separation ◽  
Latex Agglutination ◽  
Escherichia Coli O157 ◽  
High Genetic Diversity ◽  
E Coli ◽  
System A ◽  
Metabolic Genes ◽  
Beef Carcasses

The objective of the study was to characterize virulence genes and subtype Escherichia coli O157:H7 and O157:H(−) isolates obtained from a vertically integrated feedlot slaughter plant in Mexico. A total of 1,695 samples were collected from feedlots, holding pens, colon contents, hides, and carcasses. E. coli O157:H7 detection and confirmation was carried out using conventional microbiology techniques, immunomagnetic separation, latex agglutination, and the BAX system. A total of 97 E. coli O157 strains were recovered and screened for key virulence and metabolic genes using multiplex and conventional PCR. Eighty-eight (91.72%) of the strains carried stx2, eae, and ehxA genes. Ten isolates (8.25%) were atypical sorbitol-fermenting strains, and nine were negative for the flicH7 gene and lacked eae, stx1, stx2, and ehxA. One sorbitol-positive strain carried stx2, eae, tir, toxB, and iha genes but was negative for stx1 and ehxA. Pulsed-field gel electrophoresis (PFGE) analysis yielded 49 different PFGE subtypes, showing a high genetic diversity; however, the majority of the typical isolates were closely related (80 to 90% cutoff). Atypical O157 isolates were not closely related within them or to typical E. coli O157:H7 isolates. Identical PFGE subtypes were found in samples obtained from colon contents, feedlots, holding pens, and carcasses. Isolation of a sorbitol-fermenting E. coli O157 positive for a number of virulence genes is a novel finding in Mexico. These data showed that genetically similar strains of E. coli O157:H7 can be found at various stages of beef production and highlights the importance of preventing cross-contamination at the pre- and postharvest stages of processing.

Pulsed-Field Gel Electrophoresis Characterization of Shiga Toxin–Producing Escherichia coli O157 from Hides of Cattle at Slaughter

2002 ◽  
Vol 65 (7) ◽  
pp. 1172-1176 ◽  
Author(s):  
S. M. AVERY ◽  
A. SMALL ◽  
C.-A. REID ◽  
S. BUNCIC
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Gel Electrophoresis ◽  
Immunomagnetic Separation ◽  
Pulsed Field Gel Electrophoresis ◽  
Escherichia Coli O157 ◽  
Adult Cattle ◽  
Pulsed Field ◽  
E Coli ◽  
High Prevalence

Contamination of the brisket areas of the hides of healthy adult cattle with Shiga toxin–producing Escherichia coli O157 at slaughter in England was studied. In total, 73 cattle consignments comprising 584 animals delivered to one abattoir over 3 days during 1 week in July 2001 were studied: 26 cattle consignments arriving on Monday, 32 consignments arriving on Wednesday, and 15 consignments arriving on Friday. Consignment sizes ranged from 1 to 23 animals, with a mean consignment size of 8. The hide of the first animal to be slaughtered in each consignment was sampled by using a sponge swab moistened with 0.85% saline to rub an unmeasured brisket (ventral) area (ca. 30 by 30 cm). The process of isolating E. coli O157 from the swabs consisted of enrichment, screening with immunoprecipitation assay kits, and immunomagnetic separation. E. coli O157 was found on 24 of 73 (32.9%) cattle hides examined, and 21 of these 24 isolates produced Shiga toxins. The 24 E. coli O157 isolates produced six different pulsed-field gel electrophoresis profiles, and 18 (75%) of the isolates were of one prevalent clone. The high prevalence of one E. coli O157 clone on the hides of cattle at slaughter could be due to a high prevalence of that clone on the 18 farms involved (not investigated in the current study), in the postfarm transport or lairage environments, or both. Since the lairage environment, but not the farm of origin or the postfarm transport vehicle, was a factor common to all 18 cattle consignments, it could have played an important role in spreading the prevalent E. coli O157 clone to the cattle hides. Lairage pen floors and the stunning box floor were identified as the most probable sites along the unloading-to-slaughter route at which the brisket areas of cattle hides could become contaminated.

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