Prevalence and Molecular Characterization of Escherichia coli O157:H7 in Bulk Tank Milk and Fecal Samples from Cull Cows: A 12-Month Survey of Dairy Farms in East Tennessee

2002 ◽  
Vol 65 (5) ◽  
pp. 752-759 ◽  
Author(s):  
S. E. MURINDA ◽  
L. T. NGUYEN ◽  
S. J. IVEY ◽  
B. E. GILLESPIE ◽  
R. A. ALMEIDA ◽  
...  
Keyword(s):  
Shiga Toxin ◽  
Dairy Farms ◽  
Escherichia Coli O157 ◽  
Bacterial Isolates ◽  
Macconkey Agar ◽  
Bulk Tank Milk ◽  
Fecal Samples ◽  
E Coli ◽  
Milk Samples ◽  
Bulk Tank

A study on the prevalence of Escherichia coli O157:H7 was conducted on 30 dairy farms in east Tennessee between May 2000 and April 2001. This pathogen was isolated from 8 of 30 (26.7%) dairy farms at various sampling times. A total of 415 fecal samples from cull dairy cows and 268 bulk tank milk samples were analyzed. Overall, 10 of 683 (1.46%) samples (2 of 268 [0.75%] milk samples and 8 of 415 [1.93%] fecal samples) tested positive for E. coli O157:H7. Food and Drug Administration Bacteriological Analytical Manual protocols were used for the conventional isolation and confirmation of E. coli O157:H7. Samples were shake cultured (150 rpm) at 42°C for 24 h in tryptic soy broth containing 2 mg of novobiocin per liter. White colonies isolated on cefixime-tellurite sorbitol MacConkey agar plates were evaluated for fluorescence on sorbitol MacConkey agar supplemented with 0.025 g of methylumbelliferyl-β-d-glucuronide per liter. Nonfluorescing white colonies were biochemically typed and serologically confirmed. Multiplex polymerase chain reaction profiles of E. coli O157: H7 isolates indicated the presence of common virulence factors (Shiga toxin, enterohemolysin, and intimin) of Shiga toxin–producing E. coli, suggesting the potential human pathogenicity of bacterial isolates. Pulsed-field gel electrophoresis profiles of SpeI and XbaI restriction enzyme–digested genomic DNA were used to establish relatedness among bacterial isolates. Data from this study indicate that both cull dairy cows and bulk tank milk pose a potential hazard with regard to human foodborne illness. It is therefore imperative to develop on-farm and preharvest pathogen reduction programs to control the carriage of E. coli O157:H7 pathogens.

scholarly journals Detection of shiga toxin producing Escherichia coli isolated from children and cattle using PCR technique

2010 ◽  
Vol 9 (3) ◽  
pp. 76
Author(s):  
H. N. A'aiz, And F. A. Abdulla A. H. Al- Hama
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Dna Marker ◽  
Hospitalized Children ◽  
Bacterial Isolates ◽  
Macconkey Agar ◽  
Biochemical Tests ◽  
Fecal Samples ◽  
E Coli ◽  
Stx2 Gene

This study was undertaken to detect STEC isolates, gene(Stx2) in Escherichia coli isolatesand characterize them by biochemical tests , enterohemolysin production and PCR.During aperiod of seven months (November 2007 to May 2008), a total of 280 fecal samples werecollected from 120 hospitalized children suffering from diarrhea and 160 cattle fecal samples .Feces specimens were screened for the presence of NSF E. coli and STEC by cultured onsorbitol MacConkey agar (SMAC).A total of 209 (74.6%) non-sorbitol fermenting (NSF)bacterial isolates were obtained , 69 (57.5%) from children fecal samples and 140 (87.5%) fromcattle feces . Of which 5 (4.16%) NSF E. coli isolated from children fecal samples and 38(23.75%) from cattle feces. NSF isolates were identified as Shiga toxin producing E. coli(STEC), but only 16 (10%) isolates of cattle and 2 (1.6%)isolates of children were PCR-positivefor (Stx2) gene which gave amplification bands at 346 bp using DNA marker in the interpretationof the results. Among 18 STEC studied, a total of 16 (88.8%) isolates expressed enterohemolysinon washing sheep blood agar plates.On the other hand, the study was showed that the sensitivityand specificity of PCR technique in diagnosis of STEC were 41.8% , 100% respectively, incomparison with other tests like biochemical tests, sensitivity and specificity of these tests were(100% , 86.9%) respectively.

Characterization of the microbiota and resistome of bulk tank milk samples from Prince Edward Island dairy farms

2021 ◽  
Author(s):  
Landon M.C. Warder ◽  
Enrique Doster ◽  
Jennifer K. Parker ◽  
Paul S. Morley ◽  
J.T. McClure ◽  
...  
Keyword(s):  
Prince Edward Island ◽  
Dairy Farms ◽  
Bulk Tank Milk ◽  
Milk Samples ◽  
Bulk Tank

Prevalence of shiga toxin-producing Escherichia coli O157, O26 and O111 in milk, meat and faeces of cattle, sheep and pigs slaughtered in Benin

2020 ◽  
Vol 152 ◽  
pp. 15667-15675
Author(s):  
Chakirath Folakè Arikè Salifou ◽  
Cyrille Boko ◽  
Isidore Houaga ◽  
Raoul Agossa ◽  
Isabelle Ogbankotan ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Virulence Genes ◽  
Animal Origin ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Milk Samples ◽  
Food Of Animal Origin ◽  
Cattle Faeces ◽  
Risk Assessment And Management

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

scholarly journals Haptoglobin and serum amyloid A in relation to the somatic cell count in quarter, cow composite and bulk tank milk samples

2007 ◽  
Vol 74 (2) ◽  
pp. 198-203 ◽  
Author(s):  
Maria Åkerstedt ◽  
Karin Persson Waller ◽  
Åse Sternesjö
Keyword(s):  
Somatic Cell ◽  
Cell Count ◽  
Somatic Cell Count ◽  
Serum Amyloid A ◽  
Dairy Farms ◽  
Serum Amyloid ◽  
Bulk Tank Milk ◽  
Amyloid A ◽  
Milk Samples ◽  
Bulk Tank

Milk somatic cell count (SCC) is the gold standard in diagnosis of subclinical mastitis, and is also an important parameter in quality programmes of dairy cooperatives. As routine SCC analysis is usually restricted to central laboratories, much effort has been invested in the search for alternative biomarkers of mastitis and milk quality, including the presence in the milk of the acute phase proteins (APP), haptoglobin (Hp) and serum amyloid A (SAA). The aim of this study was to investigate relationships between Hp, SAA and SCC in quarter, cow composite, and bulk tank milk samples. Cows (n=165), without any clinical signs of disease or abnormalities in the milk or udder, from three different dairy farms, were used. Cow composite milk samples from all cows delivering milk at the sampling occasion were taken once in each herd. In one of the farms, representative quarter milk samples (n=103) from 26 cows were also collected. In addition, bulk tank milk samples from 96 dairy farms were included in the study. Samples were analysed for Hp, SAA and SCC, and relationships between the parameters were evaluated at quarter, cow and tank milk levels using Chi-square analysis. Milk samples were categorized according to their SCC, and the presence, or no presence, of SAA and Hp, based on the detection limits of the screening methods (0·3 mg/l and 1·0 mg/l for SAA and Hp, respectively). Hp and SAA were found in milk at quarter, cow composite and bulk tank levels. A large proportion (53%) of the animals had detectable milk concentrations of APP, and SAA was detected more frequently, and at higher concentrations than Hp, regardless of sample type. SAA was detected in as many as 82% of the bulk tank milk samples. Significant relationships were found between Hp, SAA and SCC at quarter and cow composite milk levels, but only between SAA and SCC at bulk tank milk level. Detectable levels of APP were more common at high SCC.

scholarly journals Prevalence of Coxiella burnetii in cows’ and ewes’ bulk tank milk samples from selected dairy farms of Central Italy

2017 ◽  
Vol 16 (4) ◽  
pp. 673-676 ◽  
Author(s):  
Fabrizia Guidi ◽  
Annalisa Petruzzelli ◽  
Floriana Ciarrocchi ◽  
Anna Duranti ◽  
Andrea Valiani ◽  
...  
Keyword(s):  
Coxiella Burnetii ◽  
Central Italy ◽  
Dairy Farms ◽  
Bulk Tank Milk ◽  
Milk Samples ◽  
Bulk Tank

Escherichia coli O157 and non-O157 Shiga Toxin–Producing Escherichia coli in Fecal Samples of Finished Pigs at Slaughter in Switzerland

2006 ◽  
Vol 69 (2) ◽  
pp. 260-266 ◽  
Author(s):  
M. KAUFMANN ◽  
C. ZWEIFEL ◽  
M. BLANCO ◽  
J. E. BLANCO ◽  
J. BLANCO ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Escherichia Coli O157 ◽  
Strongly Correlated ◽  
Fecal Samples ◽  
E Coli ◽  
High Prevalence ◽  
Strain Testing ◽  
Typical Epec ◽  
Uremic Syndrome

Fecal samples from 630 slaughtered finisher pigs were examined by PCR to assess the shedding of Escherichia coli O157 (rfbE) and Shiga toxin–producing E. coli (STEC, stx). The proportion of positive samples was 7.5% for rfbE and 22% for stx. By colony hybridization, 31 E. coli O157 and 45 STEC strains were isolated, and these strains were further characterized by phenotypic and genotypic traits. Among E. coli O157 strains, 30 were sorbitol positive, 30 had an H type other than H7, and none harbored stx genes. Intimin (eae), enterohemolysin (ehxA), EAST1 (astA), and porcine A/E–associated protein (paa) were present in 10, 3, 26, and 6% of strains. Among them, one eae-γ1–positive O157:H7 strain testing positive for ehxA and astA and two eae-α1–positive O157:H45 strains were classified as enteropathogenic E. coli (EPEC). The O157:H45 EPEC harbored the EAF plasmid and the bfpA gene, factors characteristic for typical EPEC. The isolated STEC strains (43 sorbitol positive) belonged to 11 O:H serotypes, including three previously reported in human STEC causing hemolytic uremic syndrome (O9:H−, O26:H−, and O103:H2). All but one strain harbored stx2e. The eae and ehxA genes, which are strongly correlated with human disease, were present in only one O103:H2 strain positive for stx1 and paa, whereas the astA gene was found more frequently (14 strains). High prevalence of STEC was found among finisher pigs, but according to the virulence factors the majority of these strains seem to be of low virulence.

Risk of Escherichia coli O157:H7, Non-O157 Shiga Toxin–Producing Escherichia coli, and Campylobacter spp. in Food Animals and Their Products in Qatar

2015 ◽  
Vol 78 (10) ◽  
pp. 1812-1818 ◽  
Author(s):  
HUSSNI O. MOHAMMED ◽  
KORANA STIPETIC ◽  
AHMED SALEM ◽  
PATRICK McDONOUGH ◽  
YUNG FU CHANG ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Cross Sectional Study ◽  
Campylobacter Coli ◽  
Escherichia Coli O157 ◽  
Cross Sectional ◽  
Fecal Samples ◽  
E Coli ◽  
Cattle Feces ◽  
Campylobacter Spp

Escherichia coli O157:H7, non-O157 E. coli, and Campylobacter spp. are among the top-ranked pathogens that threaten the safety of food supply systems around the world. The associated risks and predisposing factors were investigated in a dynamic animal population using a repeat-cross-sectional study design. Animal and environmental samples were collected from dairy and camel farms, chicken processing plants, and abattoirs and analyzed for the presence of these pathogens using a combination of bacterial enrichment and real-time PCR tests without culture confirmation. Data on putative risk factors were also collected and analyzed. E. coli O157:H7 was detected by PCR at higher levels in sheep and camel feces than in cattle feces (odds ratios [OR], 6.8 and 21.1, respectively). Although the genes indicating E. coli O157:H7 were detected at a relatively higher rate (4.3%) in fecal samples from dairy cattle, they were less common in milk and udder swabs from the same animals (1 and 2%, respectively). Among the food adulterants, E. coli O103 was more common in cattle fecal samples, whereas O26 was more common in sheep feces and O45 in camel feces compared with cattle (OR, 2.6 and 3.1, respectively). The occurrence of E. coli in the targeted populations differed by the type of sample and season of the year. Campylobacter jejuni and Campylobacter coli were more common in sheep and camel feces than in cattle feces. Most of the survey and surveillance of E. coli focused on serogroup O157 as a potential foodborne hazard; however, based on the PCR results, non-O157 Shiga toxin–producing E. coli serotypes appeared to be more common, and efforts should be made to include them in food safety programs.

scholarly journals Detection of Escherichia coli O157:H7 and Shiga toxin 2a gene in pork, pig feces, and clean water at Jagalan slaughterhouse in Surakarta, Central Java Province, Indonesia

2019 ◽  
Vol 12 (10) ◽  
pp. 1584-1590
Author(s):  
Maria Kristiani Epi Goma ◽  
Alvita Indraswari ◽  
Aris Haryanto ◽  
Dyah Ayu Widiasih
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Shiga Toxin ◽  
Water Samples ◽  
Animal Origin ◽  
Clean Water ◽  
Escherichia Coli O157 ◽  
Fecal Samples ◽  
E Coli ◽  
Pork Samples

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.

scholarly journals Molecular Ecology of Listeria monocytogenes: Evidence for a Reservoir in Milking Equipment on a Dairy Farm

2008 ◽  
Vol 75 (5) ◽  
pp. 1315-1323 ◽  
Author(s):  
Alejandra A. Latorre ◽  
Jo Ann S. Van Kessel ◽  
Jeffrey S. Karns ◽  
Michael J. Zurakowski ◽  
Abani K. Pradhan ◽  
...  
Keyword(s):  
New York ◽  
Listeria Monocytogenes ◽  
Environmental Samples ◽  
Molecular Ecology ◽  
New York State ◽  
Dairy Farm ◽  
Bulk Tank Milk ◽  
Fecal Samples ◽  
Milk Samples ◽  
Bulk Tank

ABSTRACT A longitudinal study aimed to detect Listeria monocytogenes on a New York State dairy farm was conducted between February 2004 and July 2007. Fecal samples were collected every 6 months from all lactating cows. Approximately 20 environmental samples were obtained every 3 months. Bulk tank milk samples and in-line milk filter samples were obtained weekly. Samples from milking equipment and the milking parlor environment were obtained in May 2007. Fifty-one of 715 fecal samples (7.1%) and 22 of 303 environmental samples (7.3%) were positive for L. monocytogenes. A total of 73 of 108 in-line milk filter samples (67.6%) and 34 of 172 bulk tank milk samples (19.7%) were positive for L. monocytogenes. Listeria monocytogenes was isolated from 6 of 40 (15%) sampling sites in the milking parlor and milking equipment. In-line milk filter samples had a greater proportion of L. monocytogenes than did bulk tank milk samples (P < 0.05) and samples from other sources (P < 0.05). The proportion of L. monocytogenes-positive samples was greater among bulk tank milk samples than among fecal or environmental samples (P < 0.05). Analysis of 60 isolates by pulsed-field gel electrophoresis (PFGE) yielded 23 PFGE types after digestion with AscI and ApaI endonucleases. Three PFGE types of L. monocytogenes were repeatedly found in longitudinally collected samples from bulk tank milk and in-line milk filters.

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