scholarly journals Evaluation of the Reduction of Escherichia coli O157:H7 Surrogates in Beef Ribeye Rolls at 54.4°C

2019 ◽  
Vol 3 (2) ◽  
Author(s):  
K. Habib ◽  
J. Henson ◽  
K. Tarrant ◽  
A. G. McKeith

ObjectivesE. coli infections are a primary source of gastroenteritis requiring strict cooking and handling procedures for meat producing companies as directed by the USDA. This study evaluated the reduction of Escherichia coli O157:H7 surrogates in a low temperature cook process at a local medium-large meat processor. This is one of three microbial validation projects; the other projects will investigate Clostidium perfrigens and Salmonella spp. The objective of maintaining meat quality (rare color) throughout cooking processes urges the study of low temperature cook processes to determine their efficacy in microbial control.Materials and MethodsThis study was completed in three replications each consisting of a sample size of n = 25. Four strains of Escherichia coli (American Type Culture Collection, ATCC® BAA-1427, 1428, 1429, and 1431), each approved as a surrogate for E. coli O157:H7, were used in this study. Each surrogate was grown separately. Inoculations of surrogates were prepared utilizing 800 mL of distilled water mixed with 24 g of TSB and inoculated with surrogates. The inoculations were incubated at 37°C for 24 h prior to application. The surrogates were mixed together to make a cocktail just prior to inoculation of meat. Seventy-five (25 per replication) ribeye rolls (IMPS 112-A) were removed from vacuum bags and trimmed. Initial samples were taken to determine initial microbial load prior to inoculation. The pH and temperature were taken in raw meat, after spraying with antimicrobial, and after brining. The pH and temperature of the brine was also recorded. Meat was inoculated with 90 mL of inoculum and was distributed evenly on the surface with a sponge on a stick. The inoculum was allowed to dry for 30 min prior to sampling for inoculation load. Ribeye rolls were then sampled 15 min after going through an antimicrobial spray. Samples for raw meat, initial inoculated meat, and after antimicrobial spray were taken from the surface of the ribeye roll (approximately 100 g). Following sampling, the ribeye rolls were pumped with a brine solution (sugar, salt, and proprietary ingredients) to 15%. The meat was vacuum-packaged in cook-in bags and allowed to sit in a cooler to mimic the longest period of time from packaging until it would be placed in the smokehouse. Ribeye rolls were cooked according to Appendix A at 54.4°C for 112 min, and chilled until the internal temperature was below 4.4°C. Final cooked and chilled samples were taken by cutting a 4 cm steak from the center of the roast. All samples were packaged and sent to Food Safety Net Services (FSNS) for culturing on coliform film. At FSNS 25 g of meat and 225 mL of BPW were stomached, serial dilutions were done and plated on coliform petrifilm and allowed to incubate for 24–48 h. Results were analyzed using the proc GLM procedure of SAS, determining the LSMean and StdError as well as Microsoft Excel.ResultsInitial inoculation loads after inoculation were 6.5 logs and all cooked and chilled samples had less than 1 log. Therefore, the mean log reduction was 5.1 with a standard error of 0.04 from inoculation to post-cook over the three replications.ConclusionThe results suggest that this cook method is sufficient to reduce E. coli O157:H7 in whole-muscle beef ribeye rolls. This information would be beneficial to companies looking to preserve meat quality while utilizing a low temperature cook process.

2001 ◽  
Vol 64 (8) ◽  
pp. 1122-1127 ◽  
Author(s):  
ALEX YEOW-LIM TEO ◽  
SADHANA RAVISHANKAR ◽  
CHARLES E. SIZER

The destructive effect of high pressure (615 MPa) combined with low temperature (15°C) on various strains of Escherichia coli O157:H7 and various serovars of Salmonella in grapefruit, orange, apple, and carrot juices was investigated. The three-strain cocktail of E. coli O157:H7 (SEA13B88, ATCC 43895, and 932) was found to be most sensitive in grapefruit juice (8.34-log reduction) and least in apple juice (0.41-log reductions) when pressurized at 615 MPa for 2 min at 15°C. Correspondingly, no injured survivor was detected in grapefruit and carrot juices under similar treatment conditions. No Salmonella spp. were detected in a 2-min pressure treatment (615 MPa, 15°C) of grapefruit and orange fruit juices. Except for Enteritidis, all four serovars tested in the present study have viability loss of between 3.92- and 5.07-log reductions when pressurized in apple juice at 615 MPa for 2 min at 15°C. No injured cells were recovered from grapefruit and orange juices, whereas the same treatment demonstrated reduction in numbers of Salmonella serovars Agona and Muenchen in apple juices and to a lesser extent with Typhimurium, Agona, and Muenchen in carrot juice. The present study demonstrated that low-temperature, high-pressure treatment has the potential to inactivate E. coli O157:H7 strains and different Salmonella spp. in different fruit juices.


2003 ◽  
Vol 71 (3) ◽  
pp. 1505-1512 ◽  
Author(s):  
Stuart W. Naylor ◽  
J. Christopher Low ◽  
Thomas E. Besser ◽  
Arvind Mahajan ◽  
George J. Gunn ◽  
...  

ABSTRACT Escherichia coli O157:H7 causes bloody diarrhea and potentially fatal systemic sequelae in humans. Cattle are most frequently identified as the primary source of infection, and E. coli O157:H7 generally colonizes the gastrointestinal tracts of cattle without causing disease. In this study, persistence and tropism were assessed for four different E. coli O157:H7 strains. Experimentally infected calves shed the organism for at least 14 days prior to necropsy. For the majority of these animals, as well as for a naturally colonized animal obtained from a commercial beef farm, the highest numbers of E. coli O157:H7 were found in the feces, with negative or significantly lower levels detected in lumen contents taken from the gastrointestinal tract. Detailed examination demonstrated that in these individuals the majority of tissue-associated bacteria were adherent to mucosal epithelium within a defined region extending up to 5 cm proximally from the recto-anal junction. The tissue targeted by E. coli O157:H7 was characterized by a high density of lymphoid follicles. Microcolonies of the bacterium were readily detected on the epithelium of this region by immunofluorescence microscopy. As a consequence of this specific distribution, E. coli O157:H7 was present predominately on the surface of the fecal stool. In contrast, other E. coli serotypes were present at consistent levels throughout the large intestine and were equally distributed in the stool. This is a novel tropism that may enhance dissemination both between animals and from animals to humans. The accessibility of this site may facilitate simple intervention strategies.


2008 ◽  
Vol 71 (10) ◽  
pp. 2082-2086 ◽  
Author(s):  
LUCIANO BENEDUCE ◽  
GIUSEPPE SPANO ◽  
ARI Q. NABI ◽  
FRANCESCO LAMACCHIA ◽  
SALVATORE MASSA ◽  
...  

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.


2011 ◽  
Vol 74 (1) ◽  
pp. 6-12 ◽  
Author(s):  
F. SAVOYE ◽  
P. FENG ◽  
C. ROZAND ◽  
M. BOUVIER ◽  
A. GLEIZAL ◽  
...  

Enterohemorrhagic Escherichia coli O157:H7 is an important pathogen associated with infections caused by consumption of undercooked raw meat. Sensitive and rapid detection methods for E. coli O157:H7 are essential for the meat industry to ensure a safe meat supply. This study was conducted to compare the sensitivity of the VIDAS ultraperformance E. coli test (ECPT UP) with a noncommercial real-time (RT) PCR method and the U.S. Department of Agriculture, Food Safety and Inspection Service (USDA-FSIS) reference method for detecting E. coli O157:H7 in raw ground beef. Optimal enrichment times and the efficacy of testing different types of raw meat, either as individual samples (25 g) or as composites (375 g), were examined. For 25-g samples of each type of raw ground beef tested, 6 h of enrichment was sufficient for both the VIDAS ECPT UP and RT-PCR methods, but for 375-g samples, 24 h of enrichment was required. Both the VIDAS ECPT UP and RT-PCR methods produced results similar to those obtained with the USDA-FSIS reference method after 18 to 24 h of enrichment. The primer specificity of the RT-PCR assay and the highly specific phage ligand used in the VIDAS ECPT UP for target recognition enabled the detection of low levels of E. coli O157:H7 in 25 g of various types of raw ground beef. The tests also allowed the detection of E. coli O157:H7 in composite raw ground beef and trimmings in samples of up to 375 g.


2015 ◽  
Vol 81 (6) ◽  
pp. 2063-2074 ◽  
Author(s):  
Jitendra R. Patel ◽  
Irene Yossa ◽  
Dumitru Macarisin ◽  
Patricia Millner

ABSTRACTThis study investigated the effect of a 30-cm covering of finished compost (FC) on survival ofEscherichia coliO157:H7 andSalmonellaspp. in active static and windrow composting systems. Feedstocks inoculated withE. coliO157:H7 (7.41 log CFU/g) andSalmonella(6.46 log CFU/g) were placed in biosentry tubes (7.5-cm diameter, 30-cm height) at three locations: (i and ii) two opposing sides at the interface between the FC cover layer (where present) and the feedstock material (each positioned approximately 10 cm below the pile's surface) and (iii) an internal location (top) (approximately 30 cm below the surface). On specific sampling days, surviving populations of inoculatedE. coliO157:H7 andSalmonella, genericE. coli, and coliforms in compost samples were determined.Salmonellaspp. were reduced significantly within 24 h in windrow piles and were below the detection limit after 3 and 7 days at internal locations of windrow and static piles containing FC covering, respectively. Likewise,E. coliO157:H7 was undetectable after 1 day in windrow piles covered with finished compost. Use of FC as a covering layer significantly increased the number of days that temperatures in the windrows remained ≥55°C at all locations and in static piles at internal locations. These time-temperature exposures resulted in rapid reduction of inoculated pathogens, and the rate of bacterial reduction was rapid in windrow piles. The sample location significantly influenced the survival of these pathogens at internal locations compared to that at interface locations of piles. Finished compost covering of compost piles aids in the reduction of pathogens during the composting process.


2019 ◽  
Vol 118 (2) ◽  
pp. 023
Author(s):  
Ayelén Moreno ◽  
Claudia Castellari ◽  
Alejandra Yommi ◽  
Sandra Médici ◽  
María Alejandra Pereyra

La superficie de los frutos de kiwi (Actinidia deliciosa var. Hayward) presenta una microbiota natural, que es alterada por las prácticas agrícolas utilizadas por cada productor al momento de la cosecha, el transporte, el almacenamiento y el empaque. Los objetivos de este trabajo fueron determinar la carga microbiana total y la presencia de patógenos (Escherichia coli O157:H7 y Salmonella spp) en kiwis luego de la cosecha y curado, en tres sitios diferentes del Partido de General Pueyrredón, provincia de Buenos Aires, Argentina; y optimizar la desinfección de la fruta entera con NaClO luego del almacenamiento en frío. Se cuantificaron bacterias aerobias mesófilas totales (BAMT), hongos filamentosos (HF) y levaduras (L) y se determinó presencia/ausencia de coliformes totales (CT), Escherichia coli O157:H7 y Salmonella spp. Se detectaron diferencias significativas (p≤0,01) entre las plantaciones, en los recuentos de la microbiota que afecta la calidad y vida útil de la fruta (BAMT y HF), siendo en Batán donde se halló el mayor contenido. Todos los cultivos dieron negativo para E. coli O157:H7 y Salmonella spp. Se seleccionó un método de desinfección con NaClO de concentración 300 ppm que permitió reducir la carga microbiana inicial de BAMT, HF, L y CT de la superficie de la fruta. Los resultados presentan el grado de contaminación ambiental y humana generada al finalizar la cosecha y el curado del kiwi en el Partido de General Pueyrredón, y un método sencillo para reducirla. Se destaca la ausencia de bacterias patógenas perjudiciales para la salud del consumidor.


2019 ◽  
Vol 7 (12) ◽  
pp. 634 ◽  
Author(s):  
Junliang Zhong ◽  
Xihong Zhao

Escherichia coli O157:H7 is one of the most common pathogenic bacteria that pose a threat to food safety. The aim of this study was to investigate the mechanisms of the formation of viable but non-culturable (VBNC) E. coli O157:H7 induced by low temperature (−20 °C) using RNA sequencing (RNA-Seq) transcriptomics analysis. The results of the present investigation revealed the presence of 2298 differentially expressed genes in VBNC cells, accounting for 46.03% of the total number of genes. Additionally, GO function and KEGG pathway enrichment analysis were performed to investigate the functional and related metabolic pathways of the differentially expressed genes. We found that the ion transport, protein synthesis, and protein transmembrane transport activities were significantly improved in the VBNC cells, indicating that E. coli O157:H7 cells synthesized a considerable amount of protein to maintain the levels of their functional metabolic processes and life activities in the VBNC state. In conclusion, we suggest that the increased synthesis of proteins such as SecY, FtsY, and Ffh might indicate that they are the key proteins involved in the improvement of the transmembrane transport activities in VBNC E. coli O157:H7 cells, maintaining their functional metabolism in the VBNC state and enhancing their survival ability under low temperatures.


2001 ◽  
Vol 64 (11) ◽  
pp. 1679-1689 ◽  
Author(s):  
PEGGY P. MAK ◽  
BARBARA H. INGHAM ◽  
STEVEN C. INGHAM

Time and temperature pasteurization conditions common in the Wisconsin cider industry were validated using a six-strain cocktail of Escherichia coli O157:H7 and acid-adapted E. coli O157:H7 in pH- and ∘Brix-adjusted apple cider. Strains employed were linked to outbreaks (ATCC 43894 and 43895, C7927, and USDA-FSIS-380–94) or strains engineered to contain the gene for green fluorescent protein (pGFP ATCC 43894 and pGFP ATCC 43889) for differential enumeration. Survival of Salmonella spp. (CDC 0778, CDC F2833, and CDC HO662) and Listeria monocytogenes (H0222, F8027, and F8369) was also evaluated. Inoculated cider of pH 3.3 or 4.1 and 11 or 14°Brix was heated under conditions ranging from 60°C for 14 s to 71.1°C for 14 s. A 5-log reduction of nonadapted and acid-adapted E. coli O157:H7 was obtained at 68.1°C for 14 s. Lower temperatures, or less time at 68.1°C, did not ensure a 5-log reduction in E. coli O157:H7. A 5-log reduction was obtained at 65.6°C for 14 s for Salmonella spp. L. monocytogenes survived 68.1°C for 14 s, but survivors died in cider within 24 h at 4°C. Laboratory results were validated with a surrogate E. coli using a bench-top plate heat-exchange pasteurizer. Results were further validated using fresh unpasteurized commercial ciders. Consumer acceptance of cider pasteurized at 68.1°C for 14 s (Wisconsin recommendations) and at 71.1°C for 6 s (New York recommendations) was not significantly different. Hence, we conclude that 68.1°C for 14 s is a validated treatment for ensuring adequate destruction of E. coli O157:H7, Salmonella spp., and L. monocytogenes in apple cider.


2019 ◽  
Vol 24 (1) ◽  
pp. 277-294
Author(s):  
Rocio Esperanza Patiño-Burbano ◽  
Ana Karina Carrascal ◽  
Jorge Luis Parra-Arango ◽  
José Luis Rodríguez-Bautista

Raw cow milk is considered one of the most important vehicles for pathogenic bacteria like Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes. These three bacteria are responsible for foodborne diseases. Routine microbiological methods to detect these microorganisms in cow milk can be complicated and time consuming. The aim of this work was to evaluate a method to simultaneously detect Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes in experimentally contaminated cow milk. The assessed method combined a standard microbiological culture step, using a pre-enrichment medium that favors the growth of the three focal microorganisms: SEL broth, followed by a single PCR assay. A total of 43 interference bacterial strains were used to evaluate the method’s specificity. The detection rate for the microbiological method with standard culture media was 10 UFC/mL, and that of the PCR detection, following pre-enrichment in SEL broth, was 10 UFC/mL for S. enterica and L. monocytogenes and between 1 and 5 UFC/mL for E. coli O157:H7. The PCR method showed specificity for the reference strains. Simultaneous detection by multiple PCR using SEL broth was successful for the detection of S. enterica, E. coli O157:H7, and L. monocytogenes in samples of experimentally contaminated cow milk, featuring both a high detection rate and a high specificity. This approach promises to be a feasible routine procedure when testing milk samples in industry and public health control setups.


2005 ◽  
Vol 68 (3) ◽  
pp. 551-556 ◽  
Author(s):  
SUSUMU KAWASAKI ◽  
NAOKO HORIKOSHI ◽  
YUKIO OKADA ◽  
KAZUKO TAKESHITA ◽  
TAKASHI SAMESHIMA ◽  
...  

A multiplex PCR method was developed for simultaneous detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7 in meat samples. DNA detection sensitivity for this method was 103 CFU/ml for each pathogen. When this protocol was used for the detection of each of the above pathogenic bacteria in spiked pork samples, 1 cell per 25 g of inoculated sample could be detected within 30 h. In the samples of naturally contaminated meat, Salmonella spp., L. monocytogenes, and E. coli O157:H7 were detected over the same time period. Excellent agreement was obtained for the results of multiplex PCR and the conventional culture method, which suggests that the multiplex PCR is a reliable and useful method for rapid screening of meat products for Salmonella spp., L. monocytogenes, and E. coli O157:H7 contamination.


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