Alejandro Echeverry

Validation of a lactic acid- and citric acid-based antimicrobial product for the reduction of Escherichia coli O157:H7 and Salmonella on beef tips and whole chicken carcasses.

The objectives of this study were to determine the effects of a lactic acid- and citric acid-based antimicrobial product on the reduction of Salmonella on whole broiler carcasses during processing and the reduction of Salmonella and Escherichia coli O157:H7 on beef trim. Freshly harvested broiler carcasses were inoculated with an inoculum of Salmonella strains to yield a 10(5) CFU/ml pathogen load on the surface of the carcass. The beef tips were inoculated as well with an inoculum of either E. coli O157:H7 or Salmonella to yield 10(4) CFU/100 cm(2). After 30 min for attachment, the broiler carcasses were treated with Chicxide applied for 5 s via a spray or immersed in Chicxide for 5, 10, or 20 s. Broiler carcasses were rinsed in poultry rinse bags with 400 ml of Butterfields phosphate buffer in which Salmonella was enumerated from the diluents and Butterfields phosphate. Chicxide significantly reduced Salmonella by 1.3 log CFU/ml with spray treatment and 2.3 log CFU/ml for all dip treatments. Following 30 min of attachment, the beef tips were placed into a spray cabinet with either Beefxide or sterilized water (control) and sprayed at 1 ft/2.5 s chain speed at 40 lb/in(2). The external surface of each beef tip was swabbed (100 cm(2)) to determine pathogen loads. Beefxide significantly reduced E. coli O157:H7 by 1.4 log CFU/100 cm(2) and Salmonella by 1.1 log CFU/100 cm(2) (P < 0.05) compared with the control samples.

Substantial within-Animal Diversity of Salmonella Isolates from Lymph Nodes, Feces, and Hides of Cattle at Slaughter

ABSTRACT Lymph nodes (mandibular, mesenteric, mediastinal, and subiliac; n = 68) and fecal (n = 68) and hide (n = 35) samples were collected from beef carcasses harvested in an abattoir in Mexico. Samples were analyzed for Salmonella, and presumptive colonies were subjected to latex agglutination. Of the isolates recovered, a subset of 91 was characterized by serotyping, pulsed-field gel electrophoresis (PFGE), and antimicrobial susceptibility phenotyping. Salmonella was isolated from 100% (hide), 94.1% (feces), 91.2% (mesenteric), 76.5% (subiliac), 55.9% (mandibular), and 7.4% (mediastinal) of samples. From the 87 typeable isolates, eight Salmonella enterica serotypes, including Kentucky (32.2%), Anatum (29.9%), Reading (17.2%), Meleagridis (12.6%), Cerro (4.6%), Muenster (1.1%), Give (1.1%), and Mbandaka (1.1%), were identified. S. Meleagridis was more likely (P = 0.03) to be recovered from lymph nodes than from feces or hides, whereas S. Kentucky was more likely (P = 0.02) to be recovered from feces and hides than from lymph nodes. The majority (59.3%) of the Salmonella isolates were pansusceptible; however, multidrug resistance was observed in 13.2% of isolates. Typing by PFGE revealed that Salmonella strains generally clustered by serotype, but some serotypes (Anatum, Kentucky, Meleagridis, and Reading) were comprised of multiple PFGE subtypes. Indistinguishable PFGE subtypes and, therefore, serotypes were isolated from multiple sample types, and multiple PFGE subtypes were commonly observed within an animal. Given the overrepresentation of some serotypes within lymph nodes, we hypothesize that certain Salmonella strains may be better at entering the bovine host than other Salmonella strains or that some may be better adapted for survival within lymph nodes. Our data provide insight into the ecology of Salmonella within cohorts of cattle and offer direction for intervention opportunities.

Validation of intervention strategies to control Escherichia coli O157:H7 and Salmonella typhimurium DT 104 in mechanically tenderized and brine-enhanced beef.

After three different outbreaks were linked to the consumption of nonintact meat products contaminated with Escherichia coli O157:H7, the U.S. Department of Agriculture, Food Safety and Inspection Service published notice requiring establishments producing mechanically tenderized and moisture-enhanced beef products to reassess their respective hazard analysis and critical control point system, due to potential risk to the consumers. The purpose of this study was to determine the effectiveness of different intervention strategies (lactic acid, lactic acid bacteria, and acidified sodium chlorite) to control E. coli O157:H7 and Salmonella enterica serotype Typhimurium definitive phage type 104 in mechanically tenderized and brine-enhanced beef strip loins when applied to the steaks prior to packaging and shipment for processing. After the mechanical process, translocation of E. coli O157:H7 and Salmonella Typhimurium DT 104 from the surface into the internal muscles occurred at levels between 2.0 and 4.0 log CFU/g (from an initial inoculation level of 5.0 log) after mechanical tenderization, and at levels of 1.0 to 3.0 log CFU/g after injection, with all the interventions consistently presenting lower microbial counts (P < 0.05) than did the controls. Lactic acid bacteria reduced internal E. coli O157:H7 loads 1.2 to > 2.2 log cycles, while the acidified sodium chlorite and lactic acid reduced them between 0.8 and 3.0 log, respectively. Salmonella Typhimurium DT 104 was also reduced internally after application of all interventions between 0.9 and 2.2 log. The application of antimicrobials to the steaks prior to packaging and shipment on day 0 was effective in reducing internalization of both pathogens in nonintact beef products stored for both 14 and 21 days.

Salmonella and Escherichia coli O157:H7 prevalence in cattle and on carcasses in a vertically integrated feedlot and harvest plant in Mexico.

To determine the prevalence of Salmonella and Escherichia coli O157:H7 in cattle feedlots and the impact of subsequent contamination on carcasses in a Mexican Federal Inspection Type Standards harvest facility, 250 animals were tagged and sampled in each step of the slaughter process. Samples were taken from hides and fecal grabs, and composite samples were taken from three anatomical carcass sites (hindshank, foreshank, and inside round) during the slaughter process, at preevisceration (PE), prior to entering the hot box (PHB), and after 24 h of dry chilling (DC). Additionally, 250 fecal samples were collected from the feedlot (FL), holding pens (HP), and intestinal feces (IF), and water samples were taken from the HP area. E. coli O157:H7 and Salmonella detection were carried out with the BAX System, immunomagnetic separation, and conventional methods. Overall Salmonella prevalence was 52.5%. The highest prevalence (92.4%) was found on hides, followed by feces from the HP (91.0%), FL (55.56%), PE (49.0%), IF (46.8%), and PHB (24.8%), for all sampling periods combined. The lowest prevalence of 6.0% was found after DC. The overall prevalence of E. coli O157:H7 was as follows: 11.7% for hides, 5.2% for IF, 2.7% for FL, 2.0% for HP, 0.8% for PE, 0.4% for PHB, and 0.4% for the cooler. High prevalence of Salmonella in IF and on hides present a significant risk factor for contamination by Salmonella at the different processing steps. These results serve as a warning as to the risks of contamination in meats for these pathogens and the importance of following good manufacturing practices during beef production processes.

Inhibition of Escherichia coli O157:H7 and Clostridium sporogenes in spinach packaged in modified atmospheres after treatment combined with chlorine and lactic acid bacteria.

UNLABELLED Implementation of modified atmospheric packaging (MAP) into retail produce is a less commonly practiced method due to differences among commodities and the potential growth of anaerobes. Pathogens including Escherichia coli O157:H7 have been responsible for spinach outbreaks across the United States. In this study, hurdles, including those currently used with produce safety, such as MAP and chlorine, were combined with lactic acid bacteria (LAB) to inhibit pathogens. Spinach was coinoculated with E. coli O157:H7 and Clostridium sporogenes, a surrogate for C. botulinum, and treated with water or a hurdle that included water, chlorine, and LAB. Spinach from treatments were packaged in air (traditional), oxygen (80% O₂, 20% CO₂), or nitrogen (80% N₂, 20% CO₂) and stored in a retail display case for 9 d at 4 to 7 °C. The hurdle inhibited E. coli O157:H7 and C. sporogenes compared to controls with reductions of 1.43 and 1.10 log (P < 0.05), respectively. The nitrogen atmosphere was outperformed by air and oxygen in the reduction of E. coli O157:H7 (P < 0.05) with a decrease of 0.26 and 0.15 logs. There were no significant differences among the 3 atmospheres on C. sporogenes survival. Relative to these hurdles, we also chose to evaluate the potential benefits of LAB in pathogen control. The survival of LAB in interventions demonstrates implementation of LAB into produce could control pathogens, without damaging produce or altering organoleptic properties. PRACTICAL APPLICATION The goal of our work was to identify methods that could reduce food-borne pathogens in packaged spinach products. Using current industry techniques in combination with unique methods, such as the use of beneficial bacteria, our research identified whether harmful microorganisms could be eliminated. Our data demonstrate that specific packaging conditions with beneficial bacteria can help eliminate or reduce the survival of E. coli O157:H7 and C. sporogenes (a model for C. botulinum) in produce.

Salmonella and Campylobacter baseline in retail ground beef and whole-muscle cuts purchased during 2010 in the United States.

Salmonella enterica and Campylobacter spp. cause a considerable number of human illnesses each year, and the vast majority of cases are foodborne. The purpose of this study was to establish the baseline of Salmonella and Campylobacter in beef products purchased from U.S. retail markets. Sampling was carried out in 38 American cities. Retail raw ground and whole-muscle beef (n = 2,885) samples were purchased and examined for the presence of Salmonella. Samples testing positive for Salmonella were identified with the commercial BAX System, which is a real-time PCR-based system. Of the original samples purchased, 1,185 were selected and tested for the presence of Campylobacter. Positive samples were isolated via direct plating and confirmed via agglutination and biochemical testing. Salmonella was detected in 0.66% of the total samples purchased. The prevalence of Salmonella in ground beef packages was 0.42% for modified atmosphere packaging, 0.63% for chub packaging, and 0.59% for overwrapped packages. Salmonella was detected in 1.02% of whole-muscle cuts. There was no relationship (P = 0.18) between product type (ground or whole muscle) and the percentage of positive samples. Campylobacter was recovered from 9.3% of samples. A greater percentage (17.24%, P < 0.01) of whole-muscle cuts tested positive for Campylobacter compared with ground beef samples (7.35%). Estimating pathogen baselines in U.S. retail beef is essential for allotting resources and directing interventions for pathogen control. These data can be utilized for a more complete understanding of these pathogens and their impact on public health from the consumption of beef products.

Survival of Escherichia coli O157:H7 in bovine feces over time under various temperature conditions.

Although Escherichia coli O157:H7 prevalence estimates in cattle have increased over time due to improvements in detection methods, fecal sample transport conditions from farm to microbiological laboratories for further analysis may be a factor for prevalence underestimation. The objective of this study was to compare and determine the survival characteristics of E. coli O157:H7 in bovine feces under various storage conditions that could be encountered during transport. Fecal pats were inoculated with a four-strain cocktail of antibiotic-resistant E. coli O157:H7 to contain approximately 1 x 10(5) CFU/g. Inoculated and control samples were taken after 0, 24, 48, 120, and 168 h at each storage temperature and examined for presence and numbers of E. coli O157:H7. Each sample was subdivided and placed at each of the four following temperatures: 37, 23, 4.4 degrees C, and in a plastic cooler with refrigerant packs (0, 4, 4, 21, and 23 degrees C at five sampling times, respectively) to simulate transportation conditions. A statistically significant decrease in the population of the pathogen was observed after 48 h in samples held at 37 degrees C (P < 0.01) and after 168 h at 4.4 degrees C (P = 0.02). At 37 degrees C, E. coli O157:H7 was not detected after 48 h, either by direct plating (P < 0.01) or by immunomagnetic separation. Overall, the results of this study showed that E. coli O157:H7 survived without significant detriment in bovine fecal material inside the cooler for up to 168 h. These results indicate that shipment and storage under these conditions before microbiological analysis would be acceptable and should not affect pathogen detection.

Validation of lactic acid bacteria, lactic acid, and acidified sodium chlorite as decontaminating interventions to control Escherichia coli O157:H7 and Salmonella Typhimurium DT 104 in mechanically tenderized and brine-enhanced (nonintact) beef at the purveyor.

After three different outbreaks were linked to the consumption of nonintact meat products contaminated with Escherichia coli O157:H7, the U.S. Food Safety and Inspection Service published notice requiring establishments producing mechanically tenderized and moisture-enhanced beef products to reassess their respective hazard analysis and critical control point systems, due to potential risk to the consumers. The objective of this study was to validate the use of lactic acid bacteria (LAB), acidified sodium chlorite (ASC), and lactic acid (LA) sprays when applied under a simulated purveyor setting as effective interventions to control and reduce E. coli O157:H7 and Salmonella Typhimurium DT 104 in inoculated U.S. Department of Agriculture (USDA) Choice strip loins (longissimus lumborum muscles) pieces intended for either mechanical blade tenderization or injection enhancement with a brine solution after an aging period of 14 or 21 days at 4.4°C under vacuum. After the mechanical process, translocation of E. coli O157:H7 and Salmonella Typhimurium DT 104 from the surface into the internal muscles occurred at levels between 1.00 and 5.72 log CFU/g, compared with controls. LAB and LA reduced internal E. coli O157:H7 loads up to 3.0 log, while ASC reduced the pathogen 1.4 to 2.3 log more than the control (P < 0.05), respectively. Salmonella Typhimurium DT 104 was also reduced internally 1.3 to 2.8, 1.0 to 2.3, and 1.4 to 1.8 log after application of LAB, LA, and ASC, respectively. The application of antimicrobials by purveyors prior to mechanical tenderization or enhancement of steaks should increase the safety of these types of products.

Microbial quality of condensation in fresh and ready-to-eat processing facilities

The objective was to determine the microbial risks associated with condensation in harvest, fabrication, and ready-to-eat (RTE) meat processing environments. A total of 2281 samples were collected before and during operation from areas of visible condensation, overhead pipes, and dripping pans in three plants each season during a one-year period. Significant interactions between season and plant type were observed for nearly all microorganisms, resulting in counts that were generally higher in the summer compared to other seasons. Aerobic plate counts ranged from non-detectable to 3.7log cfu100mL(-1) of condensation. Overall counts were so low that data had to be converted to results100mL(-1). Coliforms and Enterococci were not detectable in most condensation samples. Yeast and mold averaged less than 3.0log cfu100mL(-1) in all samples. Listeria spp. or Salmonella were each detected in only two samples. Condensation, present in harvest, fabrication, and RTE meat processing areas does not appear to contain microbial loads that will contaminate the product.

Diversity and susceptibility of Enterococcus isolated from cattle before and after harvest.

To investigate evidence of cross-contamination and to determine patterns of antimicrobial drug susceptibility of Enterococcus isolates in a commercial cattle processing system, samples were collected from 60 cattle shipped to a commercial abattoir. Enterococcus isolates were recovered from fecal and hide samples collected immediately before shipment from a feedlot to the abattoir, from postexsanguination hide samples at the abattoir, and from carcass samples collected after hide removal (preevisceration) and in the cooler. Of the fecal samples, 53.9% were culture positive for Enterococcus. Of hide samples collected at the feedlot, 77.8% were positive for Enterococcus, significantly lower (P < 0.01) than the proportion of hides that were culture positive at the abattoir (96.1%). For preevisceration carcass samples, Enterococcus was recovered from 58.3% of carcasses. Only 8.3% of the carcasses sampled in the cooler yielded Enterococcus. Resistance among Enterococcus isolates was common regardless of the type or location of sample from which the isolate was recovered. All 279 Enterococcus isolates were resistant to at least one antimicrobial drug, and 179 (64.2%) of these isolates were resistant to at least six drugs. The most common resistance was to chloramphenicol (100% of isolates) followed by flavomycin (90.3%), lincomycin (87.8%), tylosin (78.5%), erythromycin (76.3%), tetracycline (58.9%), quinupristin-dalfopristin (47.7%), bacitracin (17.9), streptomycin (9.0%), ciprofloxacin (1.4%), linezolid (0.7%), and salinomycin (0.4%). Enterococcus isolates also were characterized using pulsed-field gel electrophoresis to evaluate molecular similarities. Similar or indistinguishable electrophoresis patterns were found among isolates recovered at the feedlot and in the plant, providing evidence that feedlot-origin bacterial isolates are being transferred from cattle to carcasses within the processing environment through cross-contamination.