Prevalence and Antibiotic Resistance Profiles of Escherichia coli O157:H7 in Beef Products from Retail Outlets in Gaborone, Botswana

2005 ◽  
Vol 68 (2) ◽  
pp. 403-406 ◽  
Author(s):  
CLIFF A. MAGWIRA ◽  
BERHANU A. GASHE ◽  
ERNEST K. COLLISON
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Escherichia Coli O157 ◽  
Potassium Tellurite ◽  
Selective Enrichment ◽  
E Coli ◽  
Retail Outlets ◽  
Beef Products ◽  
Meat Samples ◽  
Minced Meat

Four hundred meat samples (134 meat cubes, 133 minced meat, 133 fresh sausages) were collected from 15 supermarkets and butcheries in Gaborone, Botswana, between the summer months of October 2002 and March 2003. Samples were assayed for Escherichia coli O157 by selective enrichment in modified E. coli broth containing novobiocin, followed by immuno-magnetic separation and plating onto sorbitol MacConkey agar supplemented with potassium tellurite. The isolates were biochemically and serologically confirmed by API 20E and O157 antisera, respectively. The prevalence rates for E. coli O157 were 5.22% in meat cube samples, 3.76% in minced meat samples, and 2.26% in fresh sausages. The isolates showed single, double, and triple antibiotic resistance. Fifty-three percent of them were resistant to cephalothin. Resistance was also recorded for sulphatriad (33%), colistin sulphate (26%), streptomycin (0.7%), and tetracycline (26%). It is recommended that the cause for antibiotic resistance be investigated using a larger number of samples from cattle, especially from ranching areas of the country.

Occurrence of Shiga Toxin–Producing Escherichia coli O157 in Selected Dairy and Meat Products Marketed in the City of Rabat, Morocco

2004 ◽  
Vol 67 (6) ◽  
pp. 1234-1237 ◽  
Author(s):  
N. BENKERROUM ◽  
Y. BOUHLAL ◽  
A. EL ATTAR ◽  
A. MARHABEN
Keyword(s):  
Escherichia Coli ◽  
Meat Products ◽  
Latex Agglutination ◽  
Agglutination Test ◽  
Escherichia Coli O157 ◽  
Selective Enrichment ◽  
E Coli ◽  
Meat Samples ◽  
Antigen Antibody ◽  
The City

Samples of meat and dairy products taken from the city of Rabat, Morocco, were examined for the presence of Escherichia coli O157 by the selective enrichment procedure followed by plating on cefixime–tellurite–sorbitol MacConkey agar and a latex agglutination test. The ability of isolates to produce Shiga toxins (ST1 or ST2) was also tested by an agglutination test using sensitized latex. Dairy samples (n = 44) included different products commonly consumed in the country. Meat samples (n = 36) were taken from traditional butchers because these products are generally marketed in this way. Random samples were taken from each product during the period of January through May. Of the 80 samples tested, 8 (10%) harbored E. coli O157. Four dairy and four meat samples were contaminated (9.1 and 11.1%, respectively). Of 10 E. coli O157 isolates from contaminated samples demonstrating true antigen-antibody agglutination, 5 (50%) produced either ST2 alone or ST2 plus ST1. Four of the five strains (80%) were meat isolates and produced ST2 with or without ST1, and the fifth was a dairy isolate producing ST2.

Detection, Occurrence, and Characterization of Escherichia coli O157:H7 from Raw Ewe's Milk in Spain

2006 ◽  
Vol 69 (4) ◽  
pp. 920-924 ◽  
Author(s):  
IRMA CARO ◽  
VICTOR M. FERNÁNDEZ-BARATA ◽  
ANA ALONSO-LLAMAZARES ◽  
M. ROSARIO GARCÍA-ARMESTO
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Escherichia Coli O157 ◽  
Potassium Tellurite ◽  
Selective Enrichment ◽  
E Coli ◽  
Milk Samples ◽  
Castilla Y Leon ◽  
Multiplex Pcr Assay ◽  
Ewe's Milk

A study was carried out in the Castilla y León region of Spain to investigate the presence of Escherichia coli O157:H7 in raw ewe's milk samples collected from several cheese factories during 1 year. All specimens were examined for E. coli O157:H7 by selective enrichment at 41.5 ± 1.0°C, after both 6 and 22 h of incubation, and then immunomagnetically separated and plated on cefixime–potassium tellurite–sorbitol MacConkey agar. No growth was obtained in the enrichment broth after a 6-h incubation. Presumptive colonies obtained after 22 h of incubation were screened by a multiplex PCR assay for the presence of rfbO157 and fliCH7 genes. Of all the ewe's milk samples studied, three were positive for E. coli O157:H7. The E. coli O157:H7 strains that were positive for the rfbO157 and fliCH7 genes were then analyzed by multiplex PCR for the presence of virulence genes (stx1, stx2, ehxA, and eaeA). All E. coli O157:H7 isolates were Shiga toxigenic and harbored additional genes related to virulence (ehxA and eaeA). The predominant Stx toxin type was stx2. These results demonstrate that raw ewe's milk used in cheesemaking may be sporadically contaminated with E. coli O157:H7 strains that are potentially pathogenic for humans.

Thermal Inactivation of Escherichia coli O157: H7, Salmonella senftenberg, and Enzymes with Potential as Time-Temperature Indicators in Ground Beef

1997 ◽  
Vol 60 (5) ◽  
pp. 471-475 ◽  
Author(s):  
ALICIA ORTA-RAMIREZ ◽  
JAMES F. PRICE ◽  
YIH-CHIH HSU ◽  
GIRIDARAN J. VEERAMUTHU ◽  
JAMIE S. CHERRY-MERRITT ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Thermal Inactivation ◽  
Ground Beef ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Triose Phosphate ◽  
Beef Products ◽  
Z Value ◽  
D Values ◽  
Salmonella Senftenberg

The USDA has established processing schedules for beef products based on the destruction of pathogens. Several enzymes have been suggested as potential indicators of heat processing. However, no relationship between the inactivation rates of these enzymes and those of pathogenic microorganisms has been determined. Our objective was to compare the thermal inactivation of Escherichia coli O157:H7 and Salmonella senftenberg to those of endogenous muscle proteins. Inoculated and noninoculated ground beef samples were heated at four temperatures for predetermined intervals of time in thermal-death-time studies. Bacterial counts were determined and enzymes were assayed for residual activity. The D values for E. coli O157:H7 were 46.10, 6.44, 0.43, and 0.12 min at 53, 58, 63, and 68°C, respectively, with a z value of 5.60°C. The D values for S. senftenberg were 53.00, 15.17, 2.08, and 0.22 min at 53, 58, 63, and 68°C, respectively, with a z value of 6.24°C. Apparent D values at 53, 58, 63, and 68°C were 352.93, 26.31, 5.56, and 3.33 min for acid phosphatase; 6968.64, 543.48, 19.61, and 1.40 min for lactate dehydrogenase; and 3870.97, 2678.59, 769.23, and 42.92 min for peroxidase; with z values of 7.41,3.99, and 7.80°C, respectively. Apparent D values at 53, 58, 63, and 66°C were 325.03, 60.07, 3.07, and 1.34 min for phosphoglycerate mutase; 606.72, 89.86, 4.40, and 1.28 min for glyceraldehyde-3-phosphate dehydrogenase; and 153.06, 20.13, 2.25, and 0.74 min for triose phosphate isomerase; with z values of 5.18, 4.71, and 5.56°C, respectively. The temperature dependence of triose phosphate isomerase was similar to those of both E. coli O157 :H7 and S. senftenberg, suggesting that this enzyme could be used as an endogenous time-temperature indicator in beef products.

scholarly journals Effect of Electron Beam Irradiation on Survival of Escherichia coli O157:H7 and Salmonella enterica serovar Thyphimurium in Minced Camel Meat during Refrigerated Storage

2019 ◽  
Author(s):  
A. Amiri ◽  
H. Zandi ◽  
H. Mozaffari Khosravi
Keyword(s):  
Escherichia Coli ◽  
Electron Beam ◽  
Electron Beam Irradiation ◽  
Escherichia Coli O157 ◽  
Refrigerated Storage ◽  
Beam Irradiation ◽  
E Coli ◽  
Serovar Typhimurium ◽  
Camel Meat ◽  
Meat Samples

Background: Electron beam irradiation is one of the effective ways to control foodborne pathogens. We evaluated the effect of electron beam irradiation on survival of Escherichia coli O157:H7 and Salmonella enterica serovar Thyphimurium in minced camel meat during refrigerated storage. Methods: The meat samples were inoculated with E. coli O157:H7 and S. enterica serovar Thyphimurium and then irradiated with doses of 0, 1, 2, 3, and 5 kGy. The samples were stored at 4±1 °C and evaluated microbiologically up to 10 days. Data were analyzed using SPSS software version 18. Results: The microbial loads of minced camel meat samples were significantly reduced (p<0.0001) with increasing the dose of irradiation. The most effective dose was 5 kGy that highly reduced S. enterica serovar Typhimurium, and completely destroyed E. coli O157:H7. However, E. coli O157:H7 was more sensitive to electron beam irradiation than S. enterica serovar Typhimurium. Conclusion: Electron beam irradiation effectively reduced the population of both E. coli O157:H7 and S. enterica serovar Typhimurium in minced camel meat in a dose dependent manner.

scholarly journals High Prevalence of yadA-Positive Yersinia enterocolitica in Pig Tongues and Minced Meat at the Retail Level in Finland

1999 ◽  
Vol 62 (2) ◽  
pp. 123-127 ◽  
Author(s):  
MARIA FREDRIKSSON-AHOMAA ◽  
SEBASTIAN HIELM ◽  
HANNU KORKEALA
Keyword(s):  
Yersinia Enterocolitica ◽  
Culture Method ◽  
Selective Enrichment ◽  
Conventional Culture ◽  
High Prevalence ◽  
Retail Outlets ◽  
Polymerase Chain ◽  
Meat Samples ◽  
Minced Meat ◽  
Conventional Culture Method

Polymerase chain reaction (PCR) was used to determine the prevalence of yadA-positive Yersinia enterocolitica in pig tongues and minced meat at the retail level in Finland and to confirm the yadA-positive Y. enterocolitica isolates recovered from the same samples using the conventional culture method. A total of 51 pig tongues purchased at 12 retail outlets and 255 minced meat samples purchased at 40 retail outlets in the Helsinki area were studied. The prevalence of Y. enterocolitica carrying the yadA gene was 92% in pig tongues and 25% in minced meat using PCR and 78% in tongues and 2% in minced meat with the culture method. The prevalence of yadA-positive tongues was higher (98%) when both PCR- and culture-positive results were included because Y. enterocolitica carrying the yadA gene could also be isolated in three PCR-negative tongue samples. In the minced meat samples, all PCR-negative samples were also culture-negative. With the culture method, 66 of 80 yadA-positive isolates in 38 tongues and all yadA-positive isolates (4) in four minced meat samples were recovered after selective enrichment. A total of 92 isolates of Y. enterocolitica bioserotype 4/O:3 in tongues and 5 isolates in minced meat were found, of which 13% in tongues and 20% in minced meat did not carry the yadA gene.

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.

Comparative Performance Evaluation of Real-Time PCR and Dual-Labeled Fluorescence Resonance Energy Transfer Probe-Based Melt Peak Analysis for the Detection of Escherichia coli O157:H7 in Beef Products

2019 ◽  
Vol 82 (3) ◽  
pp. 507-512
Author(s):  
JOSEPH M. BOSILEVAC ◽  
HARI P. DWIVEDI ◽  
PATRICE CHABLAIN ◽  
MICHAEL ULLERY ◽  
JOSEPH S. BAILEY ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Performance Evaluation ◽  
Energy Transfer ◽  
Real Time Pcr ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Escherichia Coli O157 ◽  
Comparative Performance ◽  
E Coli ◽  
Beef Products

ABSTRACT Contaminated beef and beef products remain a frequent vehicle for the transmission of Escherichia coli O157:H7. The current U.S. Department of Agriculture (USDA) Food Safety and Inspection Service (FSIS) regulatory testing for E. coli O157:H7 uses the method described in the USDA-FSIS Microbiology Laboratory Guidebook (MLG), chapter 5. At times, described presumptive test results are nonconfirmable, suggesting that recent PCR technological advancements and presumed enhanced sensitivity and specificity may offer beneficial changes. Here, we have evaluated the precision and sensitivity of a fluorescence resonance energy transfer–based real-time PCR assay called ECO for the detection of E. coli O157:H7. ECO detects the gene target specific to both E. coli O157:H7 and E. coli O157:non-H7 but distinguishes the two by using a melt curve analysis. A total of 3,113 O157:H7 and O157:non-H7 isolates were used to define this melting temperature–based criteria. The simulated comparative performance evaluation in the spiked beef samples indicated detection of 3 of 3 samples by ECO at &lt;3.3 log CFU/mL, whereas MLG only detected 1 of 3 (&lt;3.3 log CFU/mL). Using modified tryptic soy broth–enriched natural beef and veal product samples (n = 452), the comparative sensitivity, specificity, false-positive rate, and false-negative rate against culture between MLG and ECO were 75 versus 92%, 91 versus 99%, 8.9 versus 0.77%, and 25 versus 8.3%, respectively. Positive predictive value, negative predictive value, and the overall accuracy were found to be 56 versus 94%, 96 versus 98%, and 88 versus 98%, for MLG and ECO, respectively. These data demonstrate that the ECO assay is comparable to MLG detection of E. coli O157:H7 and offers improved sensitivity.

Occurrence and Characterization of Escherichia coli O157 and Other Serotypes in Raw Meat Products in Morocco

2008 ◽  
Vol 71 (10) ◽  
pp. 2082-2086 ◽  
Author(s):  
LUCIANO BENEDUCE ◽  
GIUSEPPE SPANO ◽  
ARI Q. NABI ◽  
FRANCESCO LAMACCHIA ◽  
SALVATORE MASSA ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Susceptibility Testing ◽  
Meat Products ◽  
Molecular Techniques ◽  
Escherichia Coli O157 ◽  
Raw Meat ◽  
E Coli ◽  
Meat Samples ◽  
The City

In this study, 100 raw meat samples were collected from 15 local Moroccan butcheries in five different areas of the city of Rabat during a period of 4 months. Overall, 7 of 15 butcheries from three areas of the city yielded strains of Escherichia coli O157. Single isolates from 9 (9%) of 100 raw meat samples were biochemically and serologically confirmed as E. coli O157. Using molecular techniques, two strains were positive for the Shiga toxin, with two additional strains containing an attaching-effacing gene. All potentially virulent serotypes isolated from these meat samples showed distinct pulsed-field gel electrophoresis profiles. Based on antibiotic susceptibility testing, more than 70% of the isolates were resistant to ampicillin and clavulanic acid–amoxicillin. Moreover, one strain was resistant to more than three antibiotics. Our study represents the first survey of E. coli O157 and related serotypes in raw meat products in Morocco.

Behavior of Escherichia coli O157:H7 during the Manufacture and Aging of Gouda and Stirred-Curd Cheddar Cheeses Manufactured from Raw Milk

2010 ◽  
Vol 73 (12) ◽  
pp. 2217-2224 ◽  
Author(s):  
DENNIS J. D'AMICO ◽  
MARC J. DRUART ◽  
CATHERINE W. DONNELLY
Keyword(s):  
Escherichia Coli ◽  
Population Level ◽  
Raw Milk ◽  
Escherichia Coli O157 ◽  
Milk Whey ◽  
Selective Enrichment ◽  
Point Counts ◽  
E Coli ◽  
Unpasteurized Milk ◽  
Low Levels

This study was conducted to examine the fate of Escherichia coli O157:H7 during the manufacture and aging of Gouda and stirred-curd Cheddar cheeses made from raw milk. Cheeses were manufactured from unpasteurized milk experimentally contaminated with one of three strains of E. coli O157:H7 at an approximate population level of 20 CFU/ml. Samples of milk, whey, curd, and cheese were collected for enumeration of bacteria throughout the manufacturing and aging process. Overall, bacterial counts in both cheese types increased almost 10-fold from initial inoculation levels in milk to approximately 145 CFU/g found in cheeses on day 1. From this point, counts dropped significantly over 60 days to mean levels of 25 and 5 CFU/g in Cheddar and Gouda, respectively. Levels of E. coli O157:H7 fell and stayed below 5 CFU/g after an average of 94 and 108 days in Gouda and Cheddar, respectively, yet remained detectable after selective enrichment for more than 270 days in both cheese types. Changes in pathogen levels observed throughout manufacture and aging did not significantly differ by cheese type. In agreement with results of previous studies, our results suggest that the 60-day aging requirement alone is insufficient to completely eliminate levels of viable E. coli O157:H7 in Gouda or stirred-curd Cheddar cheese manufactured from raw milk contaminated with low levels of this pathogen.

scholarly journals Detex for Detection of Escherichia coli O157 in Raw Ground Beef and Raw Ground Poultry

2001 ◽  
Vol 84 (3) ◽  
pp. 752-760 ◽  
Author(s):  
Yvette M Henry ◽  
Nandini Natrajan ◽  
Wendy F Lauer
Keyword(s):  
Escherichia Coli ◽  
False Positive Rate ◽  
Ground Beef ◽  
Escherichia Coli O157 ◽  
Department Of Agriculture ◽  
E Coli ◽  
Metal Plating ◽  
Positive Rate ◽  
Meat Samples ◽  
Inspection Service

Abstract A method for detection of Escherichia coli O157 in beef and poultry is presented. The method is antibody-based and uses a patented antibody-specific metal-plating procedure for the detection of E. coli O157 in enriched meat samples. Both raw ground beef and raw ground poultry were tested as matrixes for the organism. The sensitivity and specificity of the assay were 98 and 90%, respectively. The accuracy of the assay was 96%. Overall, the method agreement between the E. coli O157 Detex assay and the U.S. Department of Agriculture/Food Safety Inspection Service method was 96%. Sample temperature upon loading of the apparatus was critical to the observed false-positive rate of the system.

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