Prevalence and characterization of Listeria monocytogenes, Salmonella and Shiga toxin-producing Escherichia coli isolated from small Mexican retail markets of queso fresco.

Queso fresco (QF) is a handmade cheese consumed and produced in Latin America. In Mexico, QF production is associated with a microbiological risk. The aim of the study was to determine the incidence and characterization of Listeria monocytogenes, Salmonella spp., and Shiga toxin-producing Escherichia coli (STEC) in QF from retail markets of the north-western State of Sinaloa, Mexico, and to assess the effect of physicochemical parameters on Listeria presence. A total of 75 QF samples were obtained. L. monocytogenes, E. coli, and coliforms were detected in 9.3, 94, and 100%, respectively. Salmonella was not detected. STEC isolates showed virulence genes. Microbial loads were above the maximum values recommended by the Official Mexican Standards. Physicochemical parameters such as water activity (aw), moisture content, pH, and salinity played a role in Listeria prevalence in QF. Rigorous control in QF made in Culiacan, Mexico is needed to reduce the risk of foodborne pathogens.

Inactivation of Avian Influenza Virus Inoculated into Ground Beef Patties Cooked on a Commercial Open-Flame Gas Grill.

With the emergence of clade 2.3.4.4b H5N1 highly pathogenic avian influenza virus (AIV) infection of dairy cattle and its subsequent detection in raw milk, coupled with recent AIV infections affecting dairy farm workers, experiments were conducted to affirm the safety of cooked ground beef related to AIV because such meat is often derived from cull dairy cows. Specifically, retail ground beef (percent lean:fat = ca. 80:20) was inoculated with a low pathogenic AIV (LPAIV) isolate to an initial level of 5.6 log10 50% egg infectious doses (EID50)  per 300 g patty. The inoculated meat was pressed into patties (ca. 2.54 cm thick, ca. 300 g each) and then held at 4 °C for up to 60 min. In each of the two trials, two patties for each of the following three treatments were cooked on a commercial open-flame gas grill to internal instantaneous temperatures of 48.9 °C (120°F), 62.8 °C (145°F), or 71.1 °C (160°F), but without any dwell time. Cooking inoculated ground beef patties to 48.9 °C (ave. cooking time of ca. 15 min) resulted in a mean reduction of ≥2.5 ± 0.9 log10 EID50 per 300 g of ground beef as assessed via quantification of virus in embryonating chicken eggs (ECEs). Likewise, cooking patties on a gas grill to 62.8 °C (ave. cooking time of ca. 21 min) or to the USDA FSIS recommended minimum internal temperature for ground beef of 71.1 °C (ave. cooking time of ca. 24 min) resulted in a reduction to nondetectable levels from initial levels of ≥5.6 log10 EID50 per 300 g. These data establish that levels of infectious AIV are substantially reduced within inoculated ground beef patties (20% fat) using recommended cooking procedures.

THERMAL INACTIVATION OF CELLS OF SALMONELLA spp. IN POT PIES PREPARED WITH A BEEF, CHICKEN, OR MEAT ALTERNATIVE FILLING, WITH AND WITHOUT GRAVY, DURING COOKING IN A CONVECTION OVEN.

Cooking parameters elaborated in the U.S. Department of Agriculture's Food Safety and Inspection Service Cooking Guideline for Meat and Poultry Products (Appendix A) were evaluated for inactivation of Salmonella spp. in pot pies. To prepare dough for pot pies, flour, butter, sugar, salt, and water were mixed, portioned into balls (65 or 85 g each), flattened (ca. 13 or 15 cm diameter, ca. 0.5 cm thick), and hand pressed into pans (ca. 19.4 cm diameter). Next, a 100-g portion of beef, chicken, or a meat alternative, with or without added gravy (55 g of protein and 45 g of gravy), was inoculated with a cocktail of Salmonella spp. (ca. 6.5 or 7.9 log CFU/g of filling) and distributed onto the pie crust. After covering with a 65-g sheet of dough, pies were heat sealed in nylon polyethylene bags and stored at -20°C for up to 72 h. Frozen pot pies containing a beef or meat alternative filling were cooked in a convection oven to an internal temperature of 57.8°C (136°F) instantaneous, 62.8°C (145°F) and held for 4 min, 67.2°C (153°F) and held for 34 sec, or 71.1°C (160°F) instantaneous; whereas chicken pot pies were cooked to an internal temperature of 57.8°C (136°F) instantaneous, 62.8°C (145°F) and held for 13 min, 67.2°C (153°F) and held for 96 sec, or 73.9°C (165°F) instantaneous. Cells of Salmonella spp. were recovered from uncooked or cooked pot pies by stomaching each pie in peptone water and enumerating pathogen levels via direct plating. Cooking delivered Salmonella spp. reductions of ca. 3.6 to ≥6.3 log CFU/g of pot pie. With few exceptions, when pathogen levels decreased to below detection by direct plating (0.5 log CFU/g of pot pie), cells of Salmonella were not recovered by enrichment. In addition, there were minimal differences in the aw and moisture content of the protein filling before and after cooking, suggesting that enclosing the meat, poultry, or meat alternative filling within a dough wrapping maintains moisture in the filling during the cooking process.

Inactivation of Listeria monocytogenes and Salmonella spp. During Cooking of Country Ham and Fate of L. monocytogenes and Staphylococcus aureus During Storage of Country Ham Slices.

Thermal inactivation studies were undertaken on Listeria monocytogenes and Salmonella spp. inoculated on the surface of country ham. Hams (average = ca. 3.4 ± 0.5 kg each; average = ca. ≥18% shrinkage) were used as provided by the processor (i.e., "salted hams"), desalted in tap water (i.e., "desalted hams"), or dried for an additional period (i.e., "extra-dried hams"). Hams were surface inoculated (ca. 9.5 log CFU/ham) with a multistrain cocktail of L. monocytogenes or Salmonella spp. and cooked within a bag ina circulating water bath to an internal temperature of 130°F (54.4°C) instantaneous, 145°F (62.8°C) and held for 4 min, 153°F (67.2°C) and held for 34 s, or 160°F (71.1°C) instantaneous. Regardless of ham type, all four time and temperature combinations tested herein delivered a ≥6.7-log reduction of cells of L. monocytogenes or Salmonella spp. Differences in product pH, moisture content, or aw did not have an appreciable impact on the thermal inactivation of L. monocytogenes or Salmonella spp. on country ham. In addition, shelf-life studies were undertaken using slices of "salted" country ham that were surface inoculated (ca. 5.5 log CFU/slice) with a multistrain cocktail of L. monocytogenes or Staphylococcus aureus and then stored at 20°C. Levels of S. aureus increased by ca. ≤1.4 log CFU/slice during storage for 90 days, whereas levels of L. monocytogenes remained relatively unchanged (≤0.2 log CFU/slice increase). Our data validated that cooking parameters elaborated in the U.S. Department of Agriculture's Food Safety and Inspection Service Cooking Guideline for Meat and Poultry Products (Revised Appendix A) are sufficient to deliver significant reductions (ca. ≥6.8 log CFU/ham) in levels of L.monocytogenes and Salmonella spp. on country ham. In addition, in the event of postprocessing contamination, country ham may support the outgrowth of S. aureus or survival of L. monocytogenes during storage at 20°C for 90 days.

Fate of Listeria monocytogenes, Salmonella spp., and Shiga Toxin-Producing Escherichia coli on Slices of an All-Beef Soppressata during Storage.

Cells of Listeria monocytogenes, Salmonella spp., or Shiga toxin-producing Escherichia coli (STEC) were inoculated (ca. 4.0 log CFU/slice) onto slices (ca. 4 g each slice) of an all-beef soppressata (ca. pH 5.05 and aw 0.85). The storage of vacuum-sealed slices of inoculated soppressata at 4 °C or 20 °C for 90 days resulted in reductions of all three pathogens by ca. 2.2 to 3.1 or ca. ≥3.3 log CFU/slice, respectively. When pathogen levels decreased to below detection (≤1.18 log CFU/slice) by direct plating, it was possible to recover each of the target pathogens by enrichment, albeit more frequently from slices stored at 4 °C (p < 0.05) compared to 20 °C. In summary, the slices of the commercially produced beef soppressata selected for this study did not provide a favorable environment for either survival or outgrowth of surface-inoculated cells of L. monocytogenes, Salmonella spp., or STEC during storage.

Viability of Listeria monocytogenes and Salmonella spp. on Slices of a German-Style Bologna Containing Blends of Organic Acid Salts During Storage at 4 or 12°C.

Viability of cells of Listeria monocytogenes or Salmonella spp. was quantified on slices of a German-style bologna manufactured by a local butcher to contain no added antimicrobials or to include 0.9% or 1.3% of a blend of potassium acetate and sodium diacetate (K-Ace) or 2.5% of a blend of potassium lactate and sodium diacetate (K-Lac) as ingredients. After slicing (ca. 7.1 cm L by 6.7 cm W, ca. 0.5 cm thick, ca. 22.4 g each), a single slice of bologna was placed into a nylon-polyethylene bag and surface inoculated with 250 µL per side of a five-strain mixture of either cells of L. monocytogenes or Salmonella spp. to achieve an initial level of ca. 3.5-4.0 log CFU/slice. The packages were vacuum-sealed and then stored at 4 or 12°C for 90 and 30 days, respectively. Without antimicrobials added to the formulation, L. monocytogenes numbers increased by ca. 5.4 and 6.0 log CFU/slice at both 4 and 12°C during the entire 90- and 30-day storage period, respectively. Likewise, levels of Salmonella also increased by ca. 6.0 log CFU/slice at 12°C in the absence of added antimicrobials; however, levels of this pathogen decreased by ca. 1.7 log CFU/slice after 90 days at 4°C. With the inclusion of 0.9% or 1.3% K-Ace or 2.5% K-Lac in the bologna formulation, levels of L. monocytogenes decreased by ca. ≤0.7 log CFU/slice after 90 days at 4°C, whereas levels of Salmonella decreased by ca. 1.6-2.3 log CFU/slice. After 30 days at 12°C, levels of L. monocytogenes increased by ca. ≤3.4 log CFU/slice on product containing 0.9% K-Ace or 2.5% K-Lac but remained relatively unchanged on slices formulated with 1.3% K-Ace. For Salmonella, in the presence of 0.9% or 1.3% K-Ace or 2.5% K-Lac, pathogen levels decreased by ca. ≤0.7 log CFU/slice at 12°C after 30 days. Our data validate that the inclusion of K-Ace (0.9% or 1.3%) or K-Lac (2.5%) as ingredients is effective for controlling L. monocytogenes and Salmonella on slices of bologna during refrigerated storage.

Prevalence, characterization and sources of Listeria monocytogenes in blue crab (Callinectus sapidus) meat and blue crab processing plants.

Seven blue crab processing plants were sampled to determine the prevalence and sources of Listeria spp. and Listeria monocytogenes for two years (2006-2007). A total of 488 raw crabs, 624 cooked crab meat (crab meat) and 624 environmental samples were tested by standard methods. Presumptive Listeria spp. were isolated from 19.5% of raw crabs, 10.8% of crab meat, and 69.5% of environmental samples. L. monocytogenes was isolated from 4.5% of raw crabs, 0.2% of crab meat, and 2.1% of environmental samples. Ninety-seven percent of the isolates were resistant to at least one of the ten antibiotics tested. Eight different serotypes were found among 76 L. monocytogenes isolates tested with the most common being 4b, 1/2b and 1/2a. Automated EcoRI ribotyping differentiated 11 ribotypes among the 106 L. monocytogenes isolates. Based on ribotyping analysis, the distribution of the ribotypes in each processing plant had a unique contamination pattern. A total of 92 ApaI and 88 AscI pulsotypes among the 106 L. monocytogenes isolates were found and distinct pulsotypes were observed in raw crab, crab meat and environmental samples. Ribotypes and serotypes recovered from crab processing plants included subtypes that have been associated with listeriosis cases in other food outbreaks. Our findings suggest that molecular methods may provide critical information about sources of L. monocytogenes in crab processing plants and will augment efforts to improve food safety control strategies such as targeting specific sources of contamination and use of aggressive detergents prior to sanitizing.

Viability of Shiga Toxin-Producing Escherichia coli, Salmonella spp., and Listeria monocytogenes during Preparation and Storage of Fuet, a Traditional Dry-Cured Spanish Pork Sausage.

ABSTRACT: The primary objective of this study was to monitor viability of Shiga toxin-producing Escherichia coli (STEC), Salmonella spp., and Listeria monocytogenes during preparation and storage of fuet. Regarding methodology, coarse-ground pork (ca. 35% fat) was mixed with salt (2.5%), dextrose (0.3%), starter culture (ca. 7.0 log CFU/g), celery powder (0.5%), and ground black pepper (0.3%) and then separately inoculated with a multistrain cocktail (ca. 7.0 log CFU/g) of each pathogen. The batter was stuffed into a ca. 42-mm natural swine casing and fermented at 23 ± 2°C and ca. 95% ± 4% relative humidity to ≤pH 5.3 (≤48 h). Sausages were then dried at 12 ± 2°C and ca. 80% ± 4% relative humidity to a water activity (aw) of 0.89 (within 33 days) or aw 0.86 (within 60 days). A portion of each batch of fuet was subjected to high-pressure processing (HPP; 600 MPa for 3 min) before chubs were vacuum packaged and stored for 30 days at 20 ± 2°C. The results revealed that pathogen numbers remained relatively unchanged after fermentation (≤0.35 log CFU/g reduction), whereas reductions of ca. 0.8 to 3.2 log CFU/g were achieved after drying fuet to aw 0.89 or 0.86. Regardless of whether fuet was or was not pressure treated, additional reductions of ca. 2.2 to ≥5.3 log CFU/g after drying were achieved following 30 days of storage at 20°C. For non-HPP-treated fuet dried to aw 0.89 and stored for 30 days at 20°C, total reductions of ≥5.3 log CFU/g in levels of STEC or Salmonella spp. were achieved, whereas levels of L. monocytogenes were reduced by ca. 3.6 log CFU/g. Total reductions of ≥5.3 log CFU/g in levels of all three pathogens were achieved after drying non-HPP-treated fuet to aw 0.86. For fuet dried to aw 0.89 or 0.86, that were pressure treated and then stored for 30 days at 20°C, total reductions of >6.2 log CFU/g in levels of all three pathogens were achieved. In conclusion, the processing parameters tested herein, with or without application of HPP, validated that reductions of ≥2.0 or ≥5.0 log CFU/g in levels of STEC, Salmonella spp., and L. monocytogenes were achieved during preparation and storage of fuet.

Recovery Rate of Cells of the Seven Regulated Serogroups of Shiga Toxin-Producing Escherichia coli from Raw Veal Cutlets, Ground Veal, and Ground Beef from Retail Stores in the Mid-Atlantic Region of the United States.

ABSTRACT: A total of 482 veal cutlet, 555 ground veal, and 540 ground beef samples were purchased from retail establishments in the mid-Atlantic region of the United States over a noncontiguous 2-year period between 2014 and 2017. Samples (325 g each) were individually enriched and screened via real-time PCR for all seven regulated serogroups of Shiga toxin-producing Escherichia coli (STEC). Presumptive STEC-positive samples were subjected to serogroup-specific immunomagnetic separation and plated onto selective media. Up to five isolates typical for STEC from each sample were analyzed via multiplex PCR for both the virulence genes (i.e., eae, stx1 and/or stx2, and ehxA) and serogroup-specific gene(s) for the seven regulated STEC serogroups. The recovery rates of non-O157 STEC from veal cutlets (3.94%, 19 of 482 samples) and ground veal (7.03%, 39 of 555 samples) were significantly higher (P < 0.05) than that from ground beef (0.93%, 5 of 540 samples). In contrast, only a single isolate of STEC O157:H7 was recovered; this isolate originated from 1 (0.18%) of 555 samples of ground veal. Recovery rates for STEC were not associated with state, season, packaging type, or store type (P > 0.05) but were associated with brand and fat content (P < 0.05). Pulsed-field subtyping of the 270 viable and confirmed STEC isolates from the 64 total samples testing positive revealed 78 pulsotypes (50 to 80% similarity) belonging to 39 pulsogroups, with ≥90% similarity among pulsotypes within pulsogroups. Multiple isolates from 43 (67.7%) of 64 samples testing positive had an indistinguishable pulsotype. STEC serotypes O26 and O103 were the most prevalent serogroups in beef and veal, respectively. These findings support related findings from regulatory sampling studies over the past decade and confirm that recovery rates for the regulated STEC serogroups are higher for raw veal than for raw beef samples, as was observed in the present study of meat purchased at food retailers in the mid-Atlantic region of the United States.

Potential for growth of Clostridium perfringens from spores in pork scrapple during cooling.

We conducted stabilization studies to determine the ability of Clostridium perfringens spores to germinate and grow during exponential cooling of a commercial formulation of pork scrapple. Scrapple was inoculated with a mixture of three strains of C. perfringens spores (NTCC 8238, NCTC 8239, and ATCC 10288), vacuum packaged, and reheated (20 min/93.3 degrees C) in a circulating water bath. The cooked samples were cooled (30 s) in an ice bath before being transferred to a programmable water bath to cool through the temperature range of 54.4 degrees C to 7.2 degrees C in 12, 14, or 21 h to simulate deviations from the required cooling time of 6.5 h. After cooling, the samples, in duplicate, were analyzed to determine if growth from spores had occurred. The samples were plated onto tryptose-sulfite-cycloserine agar and incubated anaerobically at 37 degrees C for 48 h before counting the colonies. Minimal growth (less than 1.0 log) was observed during a 12- or 14 h cooling period. However, when the time to achieve 7.2 degrees C was extended to 21 h, C. perfringens spores germinated and grew from an inoculum of approximately 3.0 log(10) to approximately 7.8 log(10) CFU/g. Thus, scrapple must be cooled after cooking to 7.2 degrees C within 6.5 h, but for no more than 14 h, to prevent a food safety hazard from outgrowth of C. perfringens spores during cooling.

Inactivation of Shiga Toxin-Producing Escherichia coli in Refrigerated and Frozen Meatballs Using High Pressure Processing.

High pressure processing (HPP) was evaluated to inactivate Shiga toxin-producing Escherichia coli (STEC) in raw meatballs. Ground meat (>90% lean) was inoculated (ca. 7.0 log CFU/g) with a rifampicin-resistant cocktail of eight STEC strains (O26:H11, O45:H2, O103:H2, O104:H4, O111:H-, O121:H19, O145:NM, and O157:H7). Inoculated ground beef, ground veal, or a mixture of ground beef, pork, and veal were separately mixed with liquid whole eggs and seasonings, shaped by hand into meatballs (40 g each), and stored at -20 or at 4 °C for at least 18 h. Samples were then exposed to 400 or 600 MPa for 0 to 18 min. There were no differences (p > 0.05) in pathogen reduction related to the species of meat used or for meatballs that were refrigerated (0.9 to 2.9 log CFU/g) compared to otherwise similar meatballs that were stored frozen (1.0 to 3.0 log CFU/g) prior to HPP treatment. However, less time was needed to achieve a ≥ 2.0 log CFU/g reduction at 600 MPa (1 to 3 min) compared to 400 MPa (at least 9 min). This work provides new and practically useful information on the use of HPP to inactivate STEC in raw meatballs.

Inactivation of Shiga Toxin-Producing Escherichia coli and Listeria monocytogenes within Plant versus Beef Burgers in Response to High Pressure Processing.

ABSTRACT: We evaluated high pressure processing to lower levels of Shiga toxin-producing Escherichia coli (STEC) and Listeria monocytogenes inoculated into samples of plant or beef burgers. Multistrain cocktails of STEC and L. monocytogenes were separately inoculated (∼7.0 log CFU/g) into plant burgers or ground beef. Refrigerated (i.e., 4°C) or frozen (i.e., -20°C) samples (25 g each) were subsequently exposed to 350 MPa for up to 9 or 18 min or 600 MPa for up to 4.5 or 12 min. When refrigerated plant or beef burger samples were treated at 350 MPa for up to 9 min, levels of STEC were reduced by ca. 0.7 to 1.3 log CFU/g. However, when refrigerated plant or beef burger samples were treated at 350 MPa for up to 9 min, levels of L. monocytogenes remained relatively unchanged (ca. ≤0.3-log CFU/g decrease) in plant burger samples but were reduced by ca. 0.3 to 2.0 log CFU/g in ground beef. When refrigerated plant or beef burger samples were treated at 600 MPa for up to 4.5 min, levels of STEC and L. monocytogenes were reduced by ca. 0.7 to 4.1 and ca. 0.3 to 5.6 log CFU/g, respectively. Similarly, when frozen plant and beef burger samples were treated at 350 MPa up to 18 min, reductions of ca. 1.7 to 3.6 and ca. 0.6 to 3.6 log CFU/g in STEC and L. monocytogenes numbers, respectively, were observed. Exposure of frozen plant or beef burger samples to 600 MPa for up to 12 min resulted in reductions of ca. 2.4 to 4.4 and ca. 1.8 to 3.4 log CFU/g in levels of STEC and L. monocytogenes, respectively. Via empirical observation, pressurization did not adversely affect the color of plant burger samples, whereas appreciable changes in color were observed in pressurized ground beef. These data confirm that time and pressure levels already validated for control of STEC and L. monocytogenes in ground beef will likely be equally effective toward these same pathogens in plant burgers without causing untoward effects on product color.

Viability of Shiga Toxin-Producing Escherichia coli, Salmonella, and Listeria monocytogenes within Plant versus Beef Burgers during Cold Storage and following Pan Frying.

ABSTRACT: The viability of Shiga toxin-producing Escherichia coli (STEC), Salmonella, and Listeria monocytogenes within plant- and beef-based burgers was monitored during storage and cooking. When inoculated (ca. 3.5 log CFU/g) into 15-g portions of plant- or beef-based burgers, levels of STEC and Salmonella decreased slightly (≤0.5-log decrease) in both types of burgers when stored at 4°C, but increased ca. 2.4 and 0.8 log CFU/g, respectively, in plant-based burgers but not beef-based burgers (≤1.2-log decrease), after 21 days at 10°C. For L. monocytogenes, levels increased by ca. 1.3 and 2.6 log CFU/g in plant burgers after 21 days at 4 and 10°C, respectively, whereas pathogen levels decreased slightly (≤0.9-log decrease) in beef burgers during storage at 4 and 10°C. Regarding cooking, burgers (ca. 114 g each) were inoculated with ca. 7.0 log CFU/g STEC, Salmonella, or L. monocytogenes and cooked in a sauté pan. Cooking plant- or beef-based burgers to 62.8°C (145°F), 68.3°C (155°F), or 73.9°C (165°F) delivered reductions ranging from ca. 4.7 to 6.8 log CFU/g for STEC, ca. 4.4 to 7.0 log CFU/g for L. monocytogenes, and ca. 3.5 to 6.7 log CFU/g for Salmonella. In summary, the observation that levels of all three pathogens increased by ca. 1.0 to ca. 2.5 log CFU/g in plant-based burgers when stored at an abusive temperature (10°C) highlights the importance of proper storage (4°C) to lessen risk. However, because all three pathogens responded similarly to heat in plant-based as in beef-based burgers, well-established cooking parameters required to eliminate STEC, Salmonella, or L. monocytogenes from ground beef should be as effective for controlling cells of these same pathogens in a burger made with plant-sourced protein.

Genetic diversity and profiles of genes associated with virulence and stress resistance among isolates from the 2010-2013 interagency Listeria monocytogenes market basket survey.

Whole genome sequencing (WGS) was performed on 201 Listeria monocytogenes isolates recovered from 102 of 27,389 refrigerated ready-to-eat (RTE) food samples purchased at retail in U.S. FoodNet sites as part of the 2010-2013 interagency L. monocytogenes Market Basket Survey (Lm MBS). Core genome multi-locus sequence typing (cgMLST) and in-silico analyses were conducted, and these data were analyzed with metadata for isolates from five food groups: produce, seafood, dairy, meat, and combination foods. Six of 201 isolates, from 3 samples, were subsequently confirmed as L. welshimeri. Three samples contained one isolate per sample; mmong the 96 samples that contained two isolates per sample, 3 samples each contained two different strains and 93 samples each contained duplicate isolates. After 93 duplicate isolates were removed, the remaining 102 isolates were delineated into 29 clonal complexes (CCs) or singletons based on their sequence type. The five most prevalent CCs were CC155, CC1, CC5, CC87, and CC321. The Shannon's diversity index for clones per food group ranged from 1.49 for dairy to 2.32 for produce isolates, which were not significantly different in pairwise comparisons. The most common molecular serogroup as determined by in-silico analysis was IIa (45.6%), followed by IIb (27.2%), IVb (20.4%), and IIc (4.9%). The proportions of isolates within lineages I, II, and III were 48.0%, 50.0% and 2.0%, respectively. Full-length inlA was present in 89.3% of isolates. Listeria pathogenicity island 3 (LIPI-3) and LIPI-4 were found in 51% and 30.6% of lineage I isolates, respectively. Stress survival islet 1 (SSI-1) was present in 34.7% of lineage I isolates, 80.4% of lineage II isolates and the 2 lineage III isolates; SSI-2 was present only in the CC121 isolate. Plasmids were found in 48% of isolates, including 24.5% of lineage I isolates and 72.5% of lineage II isolates. Among the plasmid-carrying isolates, 100% contained at least one cadmium resistance cassette and 89.8% contained bcrABC, involved in quaternary ammonium compound tolerance. Multiple clusters of isolates from different food samples were identified by cgMLST which, along with available metadata, could aid in the investigation of possible cross-contamination and persistence events.

Inactivation of Toxoplasma gondii Bradyzoites after Salt Exposure during Preparation of Dry-Cured Hams.

ABSTRACT: Foodborne pathogens continue to pose a public health risk and can cause serious illness and significant outbreaks of disease in consumers. Toxoplasmosis is a zoonotic disease that occurs worldwide and is caused by the protozoan parasite, Toxoplasma gondii. The consumption of raw or undercooked infected meat, including pork, that contains infectious stages of T. gondii has been regarded as a major route of T. gondii transmission to humans. Given the occasional presence of T. gondii in pork meat, the frequent use of pork for products not intended to be cooked, such as dry-cured ham, presents a potential risk for its transmission to consumers. In this study, we investigated the viability of T. gondii in dry-cured whole hams processed using methods that were previously required for treatment of hams to inactivate Trichinella spiralis in the U.S. Code of Federal Regulations (9 CFR 318.10) and are now described in guidance documents from the U.S. Food Safety and Inspection Service. Infected pork hams were salted and cured for 33 days at 3°C and 85% ± 5% relative humidity (RH) and then were dried for up to 12 months at 12°C and 67.5% ± 2.5% RH. Inactivation of T. gondii was assessed in mouse bioassays and, serologically, by the modified agglutination test (MAT). Results showed that T. gondii bradyzoites were inactivated during the salting and curing step (33 days); no viable T. gondii was detected in the mouse bioassay and no evidence of serological conversion was detected by MAT in any of the mice inoculated with any of the samples tested during the drying step over the 12 months of the experiment. These results demonstrated that the approved protocols for production of dry-cured hams validated herein can inactivate T. gondii and lower the risk to consumers of this product.

Fate of surface-inoculated Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella typhimurium on kippered beef during extended storage at refrigeration and abusive temperatures.

The behavior of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium on kippered beef was evaluated. Individual pieces of the product were separately inoculated on the top and bottom surfaces with each three- to six-strain pathogen cocktail at ca. 6.0 log CFU per piece and stored at 4, 10, 21, or 30 degrees C for up to 28 days in each of two trials. When kippered beef was inoculated with E. coli O157:H7, Salmonella Typhimurium, or L. monocytogenes and stored at 4, 10, 21, or 30 degrees C for up to 28 days, pathogen numbers decreased ca. 0.4 to 0.9, 1.0 to 1.8, 3.0 to > or = 5.25, and > or = 5.0 to 5.25 log CFU per piece, respectively. Average D-values for E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes stored at 4 to 30 degrees C for 28 days were ca. 41 to 4.6, 40.8 to 5.3, and 29.5 to 4.3 days, respectively. As expected, the higher the storage temperature, the greater the level and rate of inactivation for all three pathogens. These data establish that kippered beef does not provide an environment conducive to proliferation of these pathogens.

Validation of cooking times and temperatures for thermal inactivation of Yersinia pestis strains KIM5 and CDC-A1122 in irradiated ground beef.

Irradiated ground beef samples (ca. 3-g portions with ca. 25% fat) inoculated with Yersina pestis strain KIM5 (ca. 6.7 log CFU/g) were heated in a circulating water bath stabilized at 48.9, 50, 52.5, 55, 57.5, or 60 degrees C (120, 122, 126.5, 131, 135.5, and 140 degrees F, respectively). Average D-values were 192.17, 34.38, 17.11, 3.87, 1.32, and 0.56 min, respectively, with a corresponding z-value of 4.67 degrees C (8.41 degrees F). In related experiments, irradiated ground beef patties (ca. 95 g per patty with ca. 25% fat) were inoculated with Y. pestis strains KIMS or CDC-A1122 (ca. 6.0 log CFU/g) and cooked on an open-flame gas grill or on a clam-shell type electric grill to internal target temperatures of 48.9, 60, and 71.1 degrees C (120, 140, and 160 degrees F, respectively). For patties cooked on the gas grill, strain KIM5 populations decreased from ca. 6.24 to 4.32, 3.51, and < or = 0.7 log CFU/g at 48.9, 60, and 71.1 degrees C, respectively, and strain CDC-A1122 populations decreased to 3.46 log CFU/g at 48.9 degrees C and to < or = 0.7 log CFU/g at both 60 and 71.1 degrees C. For patties cooked on the clam-shell grill, strain KIM5 populations decreased from ca. 5.96 to 2.53 log CFU/g at 48.9 degrees C and to < or = 0.7 log CFU/g at 60 or 71.1 degrees C, and strain CDC-A1122 populations decreased from ca. 5.98 to < or = 0.7 log CFU/g at all three cooking temperatures. These data confirm that cooking ground beef on an open-flame gas grill or on a clam-shell type electric grill to the temperatures and times recommended by the U.S. Department of Agriculture and the U.S. Food and Drug Administration Food Code, appreciably lessens the likelihood, severity, and/or magnitude of consumer illness if the ground beef were purposefully contaminated even with relatively high levels of Y. pestis.

Thermal inactivation of Escherichia coli O157:H7 in blade-tenderized beef steaks cooked on a commercial open-flame gas grill.

Beef subprimals were inoculated on the lean side with ca. 4.0 log CFU/g of a cocktail of rifampin-resistant (Rif(r)) Escherichia coli O157:H7 strains and then passed once through a mechanical blade tenderizer with the lean side facing upward. Inoculated subprimals that were not tenderized served as controls. Two core samples were removed from each of three tenderized subprimals and cut into six consecutive segments starting from the inoculated side. A total of six cores were also obtained from control subprimals, but only segment 1 (topmost) was sampled. Levels of E. coli O157:H7 recovered from segment 1 were 3.81 log CFU/g for the control subprimals and 3.36 log CFU/g for tenderized subprimals. The percentage of cells recovered in segment 2 was ca. 25-fold lower than levels recovered from segment 1, but E. coli O157:H7 was recovered from all six segments of the cores obtained from tenderized subprimals. In phase II, lean-side-inoculated (ca. 4.0 log CFU/g), single-pass tenderized subprimals were cut into steaks of various thicknesses (1.91 cm [0.75 in.], 2.54 cm [1.0 in.], and 3.18 cm [1.25 in.]) that were subsequently cooked on a commercial open-flame gas grill to internal temperatures of 48.8 degrees C (120 degrees F), 54.4 degrees C (130 degrees F), and 60 degrees C (140 degrees F). In general, regardless of temperature or thickness, we observed about a 2.6- to 4.2-log CFU/g reduction in pathogen levels following cooking. These data validate that cooking on a commercial gas grill is effective at eliminating relatively low levels of the pathogen that may be distributed throughout a blade-tenderized steak.

Thermal Inactivation of Salmonella in Pâté Made from Chicken Liver.

HIGHLIGHTS: Cooking may reduce the potential risk of salmonellosis associated with liver pâté. A 5-log reduction was achieved when inoculated pâté was cooked to an internal temperature of ≥73.8°C. A 5-log reduction was achieved when pâté was made with inoculated liver fried for >8 min at 140°C. Findings of this study may be useful for establishing cooking guidelines for liver and pâté.

Prevalence, Levels, and Viability of Salmonella in and on Raw Chicken Livers.

We surveyed chicken livers from various sources for the presence and levels of Salmonella. The pathogen was recovered from 148 (59.4%) of 249 chicken livers purchased at retail stores in Delaware, New Jersey, and Pennsylvania over about a 9-month period. Positive samples harbored Salmonella at levels of 6.4 most probable number (MPN)/g to 2.4 log CFU/g. The percentage of Salmonella-positive livers purchased at retail outlets in New Jersey (72%, 59 of 82 livers) was significantly higher (P < 0.05) than the percentage for livers purchased in Delaware (48%, 36 of 75 livers); however, this percentage was not significantly different (P > 0.05) from that for livers purchased in Pennsylvania (57.6%, 53 of 92 livers). The pathogen was also recovered more often (P = 0.019) from livers that were packaged by retailers (81 of 121 livers, 66.9%) than from livers packaged directly by processors (67 of 128 livers, 52.3%). In related studies, 12 (5.8%) of 207 chicken livers harvested from birds on a research farm tested positive for Salmonella at levels of 0.4 to 2.2 MPN/g. The recovery rate of Salmonella was 4.4% (6 of 135 livers) from livers with the gall bladder attached and 8.3% (6 of 72 livers) from livers when the gall bladder was removed at harvest on the research farm. We also quantified the levels of a nine-strain cocktail (ca. 6.5 log CFU/g) of Salmonella strains inoculated externally onto or internally into livers both before and after extended cold storage. Storage for at least 2 days at 4°C or 15 days at -20°C resulted in a decrease of about 1.0 log CFU/g in pathogen levels. Given the relatively high recovery rate (ca. 6.0 to 60.0%) and high (possibly illness causing) levels (0.4 MPN/g to 2.4 CFU/g) of Salmonella associated with chicken livers in the present study, further interventions for processors are needed to lower the prevalence and levels of this pathogen on poultry liver.