Lytic Capacity Survey of Commercial Listeria Phage Against Listeria spp. with Varied Genotypic and Phenotypic Characteristics.

Listeria monocytogenes is regularly isolated from food processing environments and is endemic in some facilities. Bacteriophage have potential as biocontrol strategies for L. monocytogenes. In this study, the lytic capacity of a commercial Listeria phage cocktail was evaluated against a library of 475 Listeria spp. isolates (426 L. monocytogenes and 49 other Listeria spp.) with varied genotypic and phenotypic characteristics. The lytic capacity of the Listeria phages was measured by spot assays where lysis was scored on a scale of 0-3 (0 = no lysis; 1 = slight lysis; 2 = moderate lysis; 3 = confluent lysis). Only 5% of all tested Listeria spp. isolates, including L. monocytogenes, were either moderately or highly susceptible (score 2 or 3) to lysis by Listeria phage when scores were averaged across temperature and phage concentration; 155 of 5700 treatment (multiplicity of infection [MOI] and temperature) and characteristic (genotype, sanitizer tolerance, and attachment capacity) combinations resulted in confluent lysis (score = 3). Odds ratios for susceptibility to lysis were calculated using multinomial logistic regression. The odds of susceptibility to lysis by phage decreased (p < 0.05) if the L. monocytogenes isolate was previously found to persist or if the phage-bacteria culture was incubated at 30°C; neither isolate persistence or temperature was significant (p ≥ 0.05) when all factors were considered. In addition, lytic efficacy varied (p < 0.05) among pulse field gel electrophoresis (PFGE) pulsotypes and may be affected by host MOI (p < 0.05). There was no effect (p > 0.05) of attachment capacity or sanitizer tolerance on phage susceptibility. This study underscores the complexity of using Listeria phage as a biocontrol for Listeria spp. in food processing facilities and highlights that phage susceptibility is most greatly impacted by genotype. Further studies are needed to evaluate these findings within a processing environment.

Evaluating the Safety of Sous-Vide Cooking for Beef Products Inoculated with Single Strains of Salmonella enterica and Escherichia coli O157.

Sous-videcooking is a growing trend among retailers and consumers. Foodborne pathogens may survive the cooking if nonvalidated parameters are used or if pathogens have enhanced thermalresistance. Pathogen inactivation from sous-vide cooking was determined when introduced directly to beef products or via contaminated spices, and with or without a finishing step. Beef products (ground beef, tenderized, and nontenderized steaks) were inoculated with pathogens (Salmonella Montevideo and Escherichia coli O157:NM) in three ways: 1) directly onto the meat 2) ground black pepper incorporated into the recipe 3) ground pepper equilibrated at 30% RH (4 d) prior to incorporation. Beef samples were vacuum-packaged and submerged in a 62.5°C water bath for 120 min. Samples were sampled at 5, 10, 20, and 120 min (recommended from a partner quality study), and a duplicate was grilled to a specific internal temperature (74°C for ground beef, 57°C for steaks) and sampled. Sous-vide cooking reduced pathogen populations by >5 log CFU/g after most treatment times, but less than grilled counterparts (ca. 1-2 log CFU/g difference; p < 0.05).There were no statistically significant differences between inoculation methods, but the tenderization of steaks resulted in significantly lower reductions of pathogens from sous-vide cooking (p < 0.05). Thisresearch challenged sous-vide cooking parameters (120 min, 62.5°C). It showed sous-vide alone lowered pathogens by >4 log CFU/g after most 20-min treatments, but 120-min sous-vide treatments or grilling would be needed for >5-log reductions.Contaminated pepper led to less consistent reductions during the cooking process, yet 2-h sous-vide still achieved a 5-log reduction. Sous-vide cooking instructions must be validated as more products and recipes are marketed.

The Use of Antimicrobial Washes to Inactivate Shiga Toxin-Producing Escherichia coli from In-Shell Pecans and Wash Water Contaminated by Different Inoculation Routes.

In-shell pecans are typically harvested after falling from trees to the ground, presenting a potential route of contamination of foodborne pathogens from soil contact. In-shell pecans are often subjected to various processing or washing steps prior to being shelled. This study determined Shiga toxin-producing Escherichia coli (STEC) reductions after treatment with antimicrobial washes on direct and soil-inoculated in-shell pecans and evaluated the cross-contamination potential of the spent pecan washes after treatment. Pecans were directly and soil-inoculated with an STEC cocktail (O157:H7, O157:NM, O121, O26). Direct inoculation was achieved by spraying the STEC cocktail on the pecans. For soil-inoculation pecans, autoclaved soil was sprayed with the STEC cocktail, homogenized for 2 min, and used to coat in-shell pecans. Inoculated pecans were washed in treatments of 2% lactic acid (LA), 1,000 ppm free chlorine (sodium hypochlorite; NaClO), hot water (HW; 85 ± 2 °C), or ambient water (C [control]; 18 ± 2 °C) for 2, 5, and 10 min and diluted to enumerate STEC populations. After treatments, 100 mL of the spent wash was vacuum filtered through a 0.45-µm membrane and plated on selective agar. HW significantly reduced STEC populations from pecans with and without soil regardless of treatment time (p < 0.05), NaClO reduced STEC populations more than the ambient control wash on directly inoculated pecans, but there were no significant differences between STEC reductions from ambient water (C), LA, and NaClO treatments on soil-inoculated pecans (p > 0.05). Larger STEC populations were enumerated from ambient water wash compared to the antimicrobial washes (p < 0.05). The HW, LA, and NaClO treatments were effective at maintaining the quality of the wash water, with STEC levels being generally at or below the detection limit (<1 CFU/100 mL), while HW was the most effective at reducing STEC from in-shell pecans with and without a soil coating (>5-log CFU/mL reductions).

Efficacy of phage therapy in pigs: systematic review and meta-analysis.

Limits on the use and efficacy of various antibiotics coupled with negative consumer perception of the practice have together spurred substantial research into compounds that could reduce the use antibiotics to control bacterial diseases in pigs. Bacteriophages are often among such potential compounds, and various groups have examined the efficacy of bacteriophages or bacteriophage products in limiting transmission or colonization of targeted bacteria. The study presented here provides a systematic review of such studies followed by a meta-analysis of aggregated data produced by each study. The data set was limited to inputs (n = 19; 576 total observations) from studies where: 1) live pigs were inoculated with a known quantity of challenge bacteria; 2) challenged animals were treated with a known quantity of phages; 3) concentrations of the challenge bacteria were measured in different tissues/fluids following phage treatment; and 4) SD (or SE to allow calculation of SD) was reported. Concentrations of challenge bacteria were significantly lower in phage-treated pigs versus challenged but untreated pigs (P < 0.0001; effect size = -1.06 1log10 colony-forming units [CFU]/g). The effect size of phage treatment was significantly greater (P < 0.05) in samples collected 48 to 96 h following phage treatment versus those collected ≤ 24 h following phage treatment. Likewise, effect size of phage treatment was significantly greater in piglets versus market-weight pigs. Across observations, phage treatment effect sizes were greatest (P < 0.01) in fecal samples versus ileal or cecal samples. Taken together, these data indicate that phage treatment can significantly reduce the concentrations of targeted bacteria in pigs; scenarios exist, however, where phage treatment could predictably be more or less effective.

Prevalence of Salmonella enterica Isolated from Food Contact and Nonfood Contact Surfaces in Cambodian Informal Markets.

ABSTRACT: The lack of hygiene and sanitation practices and insufficient infrastructure in Cambodian informal markets may increase the risk of food contamination, specifically raw vegetables, which in turn may increase the chances of contracting a foodborne disease. The aims of this study in informal markets in Cambodia were (i) to quantify the prevalence of Salmonella enterica based upon differences in season of the year (rainy versus dry), surface types (food contact surfaces versus nonfood contact surfaces), and location of vendors within the market (inside versus outside) and (ii) to characterize S. enterica serotype prevalence. A total of 310 samples were screened for S. enterica prevalence following the U.S. Department of Agriculture guidelines, and results were confirmed by PCR assay. Whole genome sequencing was used to determine the serotype for each isolate in silico using SeqSero 1.0 on draft genomes. A total of 78 samples were confirmed positive for S. enterica. During the dry season, S. enterica was more prevalent on food contact surfaces than on nonfood contact surfaces (estimated probability of detection [confidence interval]: 0.41 [0.25, 0.59] and 0.17 [0.08, 0.32], respectively; P = 0.002), but no differences were apparent in the rainy season. No differences in S. enterica prevalence were found based on location within the market (P = 0.61). Sixteen S. enterica serotypes were detected across multiple surfaces. The most common S. enterica serotypes were Rissen (18 isolates), Hvittingfoss (11), Corvallis (10), Krefeld (8), Weltevreden (6), and Altona (6). Accurate data on the prevalence of S. enterica in informal markets are crucial for the development of effective surveillance and implementation of suitable intervention strategies at the domestic level, thus preventing foodborne illness.

Investigating Salmonella enterica, Escherichia coli, and Coliforms on Fresh Vegetables Sold in Informal Markets in Cambodia†.

ABSTRACT: Vegetables in Cambodia are commonly sold in informal markets lacking food safety standards and controls. Current data on microbial contamination of vegetables in Cambodian informal markets are limited. The purpose of this study was to investigate Salmonella enterica and indicator organisms (Escherichia coli and coliforms) on the surface of fresh vegetables sold in informal markets in Cambodia. Samples of loose-leaf lettuce, tomatoes, and cucumbers were collected from 104 vendors at four informal markets in Battambang and Siem Reap provinces during the rainy and dry seasons. Detection methods for S. enterica were adapted from the U.S. Food and Drug Administration's Bacteriological Analytical Manual. Coliform and E. coli populations were quantified by plating onto E. coli/coliform count plates. S. enterica was most prevalent on lettuce during the dry season (56.5%, 95% confidence interval [CI] [41.0, 70.8]) than during the rainy season (15.4%, 95% CI [7.5, 29.1]), whereas no significant seasonal differences were apparent for tomatoes and cucumbers. Regardless of season, levels of S. enterica were highest on lettuce (5.7 log CFU/g, 95% CI [5.5, 5.9]), relative to cucumbers (4.2 log CFU/g, 95% CI [3.8, 4.6]) and tomatoes (4.3 log CFU/g, 95% CI [4.1, 4.6]). For E. coli, prevalence was higher during the rainy season (34.0%, 95% CI [25.4, 43.8]) than during the dry season (9.1%, 95% CI [4.9, 16.5]), with the highest prevalence estimated on lettuce. Coliform levels on lettuce and tomatoes were greater during the rainy season (6.3 and 5.3 log CFU/g, 95% CI [5.7, 6.8] and [4.7, 5.8], respectively) than during the dry season (5.2 and 3.9 log CFU/g, 95% CI [4.7, 5.7] and [3.4, 4.4], respectively). These results indicate seasonal patterns for microbial prevalence in lettuce and an overall high level of bacterial contamination on raw vegetables sold in Cambodian informal markets.