Modeling the Temperature Effect on the Growth of Uropathogenic Escherichia coli in Sous-Vide Chicken Breast.

Uropathogenic Escherichia coli (UPEC) is known to cause 65-75% of human urinary tract infection (UTI) cases. Poultry meat is a reservoir of UPEC, which is suspected to cause foodborne UTIs. In the present study, we aimed to determine the growth potential of UPEC in ready-to-eat chicken breasts prepared by sous-vide processing. Four reference strains isolated from the urine of UTI patients (Bioresource Collection and Research Center [BCRC] 10,675, 15,480, 15,483, and 17,383) were tested by polymerase chain reaction assay for related genes to identify their phylogenetic type and UPEC specificity. A cocktail of these UPEC strains was inoculated into sous-vide cooked chicken breast at 103-4 colony-forming unit (CFU)/g and stored at 4°C, 10°C, 15°C, 20°C, 30°C, and 40°C. Changes in the populations of UPEC during storage were analyzed by a one-step kinetic analysis method using the U.S. Department of Agriculture [USDA] Integrated Pathogen Modeling Program-Global Fit [IPMP-Global Fit]. The results showed that the combination of the no lag phase primary model and the Huang square-root secondary model fitted well with the growth curves to obtain the appropriate kinetic parameters. This combination for predicting UPEC growth kinetics was further validated using it to study additional growth curves at 25°C and 37°C, which showed that the root mean square error, bias factor, and accuracy factor were 0.49-0.59 (log CFU/g), 0.941-0.984, and 1.056-1.063, respectively. In conclusion, the models developed in this study are acceptable and can be used to predict the growth of UPEC in sous-vide chicken breast.

Surveillance of Vibrio parahaemolyticus pathogens recovered from ready-to-eat foods.

This study examined the occurrence of V. parahaemolyticus from ready-to-eat (RTE) food in Delta State, Nigeria. It also characterized antibiotic resistance and virulence gene profile patterns to determine the associated health risk hazard. Food samples total of 380 were collected randomly and assessed for V. parahaemolyticus. V. parahaemolyticus isolates were characterized for their virulence and antibiogram potentials using a phenotypic and polymerase chain reaction (PCR) approach. A total of 42 (11.1%) samples were contaminated with V. parahaemolyticus. In 17/42 (40.5%) of the V. parahaemolyticus-positive samples, the densities were < 10 MPN/g. However, 19/42 (45.2%) and 6/42 (14.3%) of the samples had densities of 10 - 102 and > 102 MPN/g, respectively. A total of 67 V. parahaemolyticus isolates were identified using PCR; 54(80.6%) isolates were multidrug resistant. A total of 22 (32.8%), 39 (58.2%), and 67 (100%) of the V. parahaemolyticus harbored the tdh, trh, and tlh toxin genes, respectively. The T3SS1 gene (vcrD1) was detected in 67 (100%) of the isolates. The T3SS2α genes which were vcrD2, vopB2, and vopT were detected in 21 (31.3%), 11 (16.4%) and 30 (44.8%) of the isolates respectively. Some of the V. parahaemolytics strains harbored the orf8 gene 20 (29.9%), and a combination of orf8 + tdh genes 12 (17.9%), categorized as pandemic strains. The antibiotic resistance genes detected in this study include blaTEM 33 (49.3), tetM 19 (28.4), cmlA 32(47.8) and sul1 14 (20.9). The concentration levels and prevalence of V. parahaemolyticus in RTE foods indicate contamination of ready-to-eat foods, particularly street foods consumed in the Delta State of Nigeria, threatening public health and consumer safety.

Characterization of virulence factors of Salmonella isolated from human stools and street food in urban areas of Burkina Faso.

BACKGROUND: This study was undertaken to identify and functionally characterize virulence genes from Salmonella isolates in street food and stool cultures. From February 2017 to May 2018, clinical and food Salmonella strains were isolated in three regions in Burkina Faso. Salmonella was serotyped according to the White-Kauffmann-Le Minor method, and polymerase chain reaction (PCR) was used to detec invA, spvR, spvC, fimA and stn virulence genes commonly associated with salmonellosis in Sub-Saharan Africa. RESULTS: A total of 106 Salmonella isolates (77 human stools; 14 sandwiches) was analyzed using a serological identification with an O-group test reagent. The presence of Salmonella was confirmed in 86% (91/106) of the samples were reactive (OMA-positive/OMB-positive). Salmonella serogroup O:4,5 was the most common serogroup detected (40%; 36/91). Salmonella Enteritidis and Typhimurium represented 5.5% (5/91) and 3.3% (3/91), respectively and were identified only from clinical isolates. Furthermore, 14 serotypes of Salmonella (12/91 human strains and 2/15 sandwich strains) were evocative of Kentucky/Bargny serotype. For the genetic profile, 66% (70/106) of the Salmonella had invA and stn genes; 77.4% (82/106) had the fimA gene. The spvR gene was found in 36.8% (39/106) of the isolates while 48.1% (51/106) had the spvC gene. Among the identified Salmonella Enteritidis and Salmonella Typhimurium isolated from stools, the virulence genes detected were invA (3/5) versus (2/3), fimA (4/5) versus (3/3), stn (3/5) versus (2/3), spvR (4/5) versus (2/3) and spvC (3/5) versus (2/3), respectively. CONCLUSION: This study reports the prevalence of Salmonella serotypes and virulence genes in clinical isolates and in street foods. It shows that food could be a significant source of Salmonella transmission to humans. Our results could help decision-making by the Burkina Faso health authority in the fight against street food-related diseases, in particular by training restaurateurs in food hygiene.

Epidemiological investigations identified an outbreak of Shiga toxin-producing Escherichia coli serotype O26:H11 associated with pre-packed sandwiches.

In October 2019, public health surveillance systems in Scotland identified an increase in the number of reported infections of Shiga toxin-producing Escherichia coli (STEC) O26:H11 involving bloody diarrhoea. Ultimately, across the United Kingdom (UK) 32 cases of STEC O26:H11 stx1a were identified, with the median age of 27 years and 64% were male; six cases were hospitalised. Among food exposures there was an association with consuming pre-packed sandwiches purchased at outlets belonging to a national food chain franchise (food outlet A) [odds ratio (OR) = 183.89, P < 0.001]. The common ingredient identified as a component of the majority of the sandwiches sold at food outlet A was a mixed salad of Apollo and Iceberg lettuce and spinach leaves. Microbiological testing of food and environmental samples were negative for STEC O26:H11, although STEC O36:H19 was isolated from a mixed salad sample taken from premises owned by food outlet A. Contamination of fresh produce is often due to a transient event and detection of the aetiological agent in food that has a short-shelf life is challenging. Robust, statistically significant epidemiological analysis should be sufficient evidence to direct timely and targeted on-farm investigations. A shift in focus from testing the microbiological quality of the produce to investigating the processes and practices through the supply chain and sampling the farm environment is recommended.

Antibiotic resistance in the pathogenic foodborne bacteria isolated from raw kebab and hamburger: phenotypic and genotypic study.

BACKGROUND: In recent years, interest in the consumption of ready-to-eat (RTE) food products has been increased in many countries. However, RTE products particularly those prepared by meat may be potential vehicles of antibiotic-resistance foodborne pathogens. Considering kebab and hamburger are the most popular RTE meat products in Iran, this study aimed to investigate the prevalence and antimicrobial resistance of common foodborne pathogens (Escherichia coli, Salmonella spp., Staphylococcus aureus, and Listeria monocytogenes) in raw kebab and hamburger samples collected from fast-food centers and restaurants. Therefore, total bacterial count (TBC), as well as the prevalence rates and antibiogram patterns of foodborne pathogens in the samples were investigated. Also, the presence of antibiotic-resistance genes (blaSHV, blaTEM, blaZ, and mecA) was studied in the isolates by PCR. RESULTS: The mean value of TBC in raw kebab and hamburger samples was 6.72 ± 0.68 log CFU/g and 6.64 ± 0.66 log CFU/g, respectively. E. coli had the highest prevalence rate among the investigated pathogenic bacteria in kebab (70%) and hamburger samples (48%). Salmonella spp., L. monocytogenes, and S. aureus were also recovered from 58, 50, and 36% of kebab samples, respectively. The contamination of hamburger samples was detected to S. aureus (22%), L. monocytogenes (22%), and Salmonella spp. (10%). In the antimicrobial susceptibility tests, all isolates exhibited high rates of antibiotic resistance, particularly against amoxicillin, penicillin, and cefalexin (79.66-100%). The blaTEM was the most common resistant gene in the isolates of E. coli (52.54%) and Salmonella spp. (44.11%). Fourteen isolates (23.72%) of E. coli and 10 isolates (29.41%) of Salmonella spp. were positive for blaSHV. Also, 16 isolates (55.17%) of S. aureus and 10 isolates (27.27%) of L. monocytogenes were positive for mecA gene. CONCLUSIONS: The findings of this study showed that raw kebab and hamburger are potential carriers of antibiotic-resistance pathogenic bacteria, which can be a serious threat to public health.

High prevalence of multidrug-resistant Escherichia coli and first detection of IncHI2/IncX4-plasmid carrying mcr-1 E. coli in retail ready-to-eat foods in China.

Antibiotic-resistant bacteria in food pose an important threat to public health. Multidrug-resistant strains in ready-to-eat (RTE) foods can be transferred to humans through diet, which increases their health risk. This study systematically investigated antibiotic resistance and antibiotic resistance genes in E. coli isolated from retail RTE foods and characterized plasmid-mediated colistin-resistant E. coli strains. A total of 1118 RTE food samples were collected from markets in 39 cities in China, and 126 E. coli strains, >95% of which were multidrug-resistant, were isolated. The isolates showed a high prevalence of resistance to tetracycline (95.24%), ampicillin (82.54%), trimethoprim-sulfamethoxazole (77.78%), nalidixic acid (74.60%), cephalothin (72.22%), chloramphenicol (66.67%), and streptomycin (53.97%). Twenty-two extended-spectrum ß-lactamase (ESBL)-producing E. coli and four colistin-resistant E. coli were identified. The resistance genes TEM, CTX-M, tetA, sul2, strA/strB, aadA, and qnrS were the most frequently detected. CTX-M-55 and CTX-M-14 were the predominant CTX-M types. All the four colistin-resistant E. coli isolates were positive for mcr-1. The mcr-1 gene can be transferred to E. coli C600 through conjugation and transformation. Whole-genome sequencing revealed that the mcr-1 genes were found in IncX4 and IncHI2 plasmids. To the best of our knowledge, this is the first report of IncHI2/IncX4 plasmid-bearing mcr-1-positive E. coli strains in RTE foods sold in markets, and the first report of the isolation of the international epidemic E. coli clone ST101 and mcr-1-carrying ESBL-producing E. coli from RTE foods. These results provide valuable information for assessing antibiotic-resistant E. coli infections and controlling antibiotic-resistant E. coli.

Quantitative risk assessment of Listeria monocytogenes in ready-to-eat (RTE) cooked meat products sliced at retail stores in Greece.

A quantitative microbial risk assessment (QMRA) model predicting the listeriosis risk related to the consumption of Ready- To- Eat (RTE) cooked meat products sliced at retail stores in Greece was developed. The probability of illness per serving assessed for 87 products available in the Greek market was found highly related to the nitrite concentration; products having a lower concentration showed a higher risk per serving. The predicted 95th percentiles of the annual listeriosis cases totaled 33 of which 13 cases were <65 years old and 20 cases ≥65 years old. The highest number of cases was predicted for mortadella, smoked turkey, boiled turkey and parizer, which were the most frequently consumed product categories. Two scenarios for assessing potential interventions to reduce the risk were tested: setting a use-by date of 14 days (these products have no use-by date based on current European Union legislation) and improving the temperature control during domestic storage. The two scenarios resulted in a decrease of the 95th and 99th percentiles of the total annual cases by 97% and 88%, respectively.

Lactic fermentation of cooked, comminuted mussel, Perna canaliculus.

The endogenous microflora of mussels, filter feeders, can include pathogens with resulting food safety concerns. The aim was to develop a cook-then-ferment technology to extend shelf life and safety of a ready-to-eat mussels. Only after cooking to destroy the mussel's endogenous microflora could an edible product be made as determined by pH decline after fermentation and the fate of common pathogens. Perna canaliculus was bought live at retail on many dates. Fermentation was with commercial lactic cultures incubated under vacuum at 30 °C for four days. Using one culture containing Pediococcus pentosaceus and Staphylococcus carnosus as a model, pH typically declined to 4.5 to 4.7, and common pathogens, Staphylococcus aureus, Salmonella and Vibrio parahaemolyticus were absent or reduced to acceptable levels. The fate of Listeria monocytogenes was studied with five cultures. These were variably effective at inhibition with one clear success, Chr Hansen's T-SC-150 containing a specific strain of Lactobacillus sakei, and flavour-generating Staphylococcus carnosus. This culture's efficacy was confirmed with sterile extracts of LAB challenging L. monocytogenes in vitro. This culture was also the most rapid fermenter by pH fall. Cook-then-ferment technology may be applied to other novel foods to minimise a disruptive endogenous microflora.

Presence of Listeria monocytogenes in Ready-to-Eat Meat Products Sold at Retail Stores in Costa Rica and Analysis of Contributing Factors.

ABSTRACT: Listeria monocytogenes is a pathogenic bacterium associated with ready-to-eat (RTE) meat products sold at the retail level. The objective of this research was to determine the prevalence of L. monocytogenes in RTE meat products sold at retail in Costa Rica and to study the factors associated with the levels of contamination; analyzed factors include hygienic practices within stores (cutting techniques and microbial contamination of products) and the behavior of the isolates (persistence against antimicrobials and transfer potential). A total of 190 samples of RTE meat products were collected and analyzed for the presence of coliforms and Listeria spp. Isolates of L. monocytogenes were then evaluated in terms of resistance to disinfectants (quaternary ammonium compound [QAC] and chlorine) and their transfer potential from food contact surfaces (knife and cutting boards). Overall Listeria spp. prevalence was 37.4% (71 of 190); Listeria innocua was present in 32.1% (61 of 190) of the products, and L. monocytogenes was found in just 2.6% (5 of 190) of the samples. Most contaminated samples were cut with a knife at the moment of purchase (44.2%). When analyzing practices within the stores, it was observed that L. monocytogenes transfer from inoculated knife to salchichón was higher for samples cut at the beginning of the experiment. In addition, L. monocytogenes transfer from inoculated cutting boards was independent of the number of slices but contamination from plastic was higher than wood. Regarding L. monocytogenes resistance to disinfectants, average reductions of 2.6 ± 1.1 log CFU/mL were detected after 6 min of exposure to 200 ppm of chlorine; however, chlorine resistance varied among the strains. Prevalence of L. monocytogenes in RTE meat products sold at retail could be associated with handling practices within the stores; further studies are necessary to estimate the impact of these practices on the overall risk for consumers.

Development and validation of a regression model for Listeria monocytogenes growth in roast beefs.

Food business operators are responsible for food safety and assessment of shelf lives for their ready-to-eat products. For assisting them, a customized software based on predictive models, ListWare, is being developed. The aim of this study was to develop and validate a predictive model for the growth of Listeria monocytogenes in sliced roast beef. A challenge study was performed comprising 51 different combinations of variables. The growth curves followed the Baranyi and Roberts model with no clear lag phase and specific growth rates in the range <0.005-0.110 hr-1. A linear regression model was developed based on 528 observations and had an adjusted R-square of 0.80. The significant predictors were storage temperature, sodium lactate, interactions between sodium acetate and temperature, and MAP packaging and temperature. The model was validated in four laboratories in three countries. For conditions where the model predicted up to + log 2 cfu/g Listeria concentration, the observed concentrations were true or below the predicted concentration in 90% of the cases. For the remaining 10%, the roast beef was coated with spices and therefore different from the others. The model will be implemented in ListWare web-application for calculation of "Listeria shelf life".

Multidrug-, methicillin-, and vancomycin-resistant Staphylococcus aureus isolated from ready-to-eat meat sandwiches: An ongoing food and public health concern.

Methicillin-resistant S. aureus (MRSA) and their antimicrobial resistance pose exacerbating global health threats and endangering everyone. Thus, the prevalence, molecular characterization of virulence genes, and antimicrobial resistance patterns of strains isolated from 225 beef burger and hot dog sandwiches vended in Mansoura city, Egypt were determined. 83.1% of the sandwiches tested were contaminated with coagulase-positive S. aureus, with a mean count of 4 × 103 CFU/g. Genes encoding mecA, α-hemolysin, staphylococcal enterotoxins, and toxic shock syndrome toxin-1 were detected in 22.6%, 96.3%, 61.1%, and 0% of the strains isolated, respectively. Of the 190 coagulase-positive strains, 43 (22.6%) were confirmed as MRSA. Among them, 4 strains (2.1%) were vancomycin-resistant S. aureus (VRSA) and resistant to all antimicrobials tested. Interestingly, all isolates were resistant to at least one of the antimicrobials tested, with 75.2% being multi-drug resistant (MDR) and an average multiple antimicrobial resistance (MAR) index of 0.503. Not less important, 100%, 96.3%, 90.5%, 79.5%, 73.7%, 62.6%, and 48.9% of isolates were resistant to Kanamycin, Nalidixic acid, Cefotaxime, Sulphamethoxazole-Trimethoprim, Penicillin G, Tetracycline, and Cephalothin, respectively. The potential hazard of MDR-, MRSA-, and VRSA-contaminated sandwiches may be an indication of the presence of what is more dangerous. Hence, strict hygienic measures and good standards of food handler's personal hygiene to prevent transmission of these pathogens to consumers are imperative.

Survival and growth of Listeria monocytogenes and Staphylococcus aureus in ready-to-eat Mediterranean vegetable salads: Impact of storage temperature and food matrix.

This study aimed to evaluate the survival or growth behavior of L. monocytogenes and S. aureus in ready-to-eat vegetable salads including Arabic salad (tomato-cucumber salad) with or without 0.5% (w/w) salt and 1% (v/w) lemon juice, tahini salad, coleslaw salad, toum sauce salad (aioli) stored at 4, 10, and 24 °C for 5 d. L. monocytogenes showed greater growth or survival in all types of salads at all temperatures tested. L. monocytogenes grew in Arabic salad without additives at 10 °C and tahini salad at 4 and 10 °C. L. monocytogenes survived all preparation and storage conditions used in the current study and its numbers were detectable in all types of salads tested, and with a maximum reduction of 3.0 log CFU/g, except for toum sauce salad at 24 °C, the cells were not detected using enrichment broth. In addition, S. aureus survived in Arabic salad with or without additives and in tahini salad with a maximum reduction of 2.3 log CFU/g. However, S. aureus numbers sharply decreased in coleslaw and toum sauce salad and at 5 d they reached undetectable levels (≥2 log CFU/g) in coleslaw at 24 °C and in toum sauce at 10 and 24 °C. Addition of lemon juice and salt to Arabic salad significantly reduced the numbers of both pathogens.

Evolution of fungal community associated with ready-to-eat pineapple during storage under different temperature conditions.

The international market of fresh-cut products has witnessed dramatic growth in recent years, stimulated by consumer's demand for healthy, nutritious and convenient foods. One of the main challenging issues for the quality and safety of these products is the potential microbial spoilage that can significantly reduce their shelf-life. The complete identification of fresh-cut product microbiota together with the evaluation of environmental factors impact on microbial composition is of primary importance. We therefore assessed the fungal communities associated with the spoilage of ready-to-eat (RTE) pineapple using a metagenetic amplicon sequencing approach, based on the ITS2 region. Our results revealed a significant variability on fungal species composition between the different batches of RTE pineapple. The initial microbiota composition was the main influencing factor and determined the progress of spoilage. Temperature and storage time were the secondary factors influencing spoilage and their impact was depending on the initial prevalent fungal species, which showed different responses to the various modifications. Our results strongly suggest that further large-scale sampling of RTE pineapple production should be conducted in order to assess the full biodiversity range of fungal community involved in the spoilage process and for unravelling the impact of important environmental factors shaping the initial microbiota.

Prevalence, virulence genes, and genetic diversity of Bacillus cereus isolated from convenience food.

BACKGROUND: In times when there is a growing interest in ready-to-eat food (RTEF), the presence of potentially pathogenic bacteria, including the toxigenic psychrotolerant bacilli from the B. cereus group, on this type of carrier may pose a real threat to the health of consumers. A significant part of RTEF is represented by vegetable products and food products made with them. The increased production of convenience foods has resulted in their international turnover growing. When coupled with a rising percentage of persons from risk groups (YOPI), including the elderly or immunocompromised, this may mean increased health risks posed by the so-called "novel pathogens", like the toxigenic psychrotolerant B. cereus sensu lato. METHODS: Food samples were analyzed for the presence and count of putative B. cereus according to the Polish Standard PN-EN ISO 7932:2005. All genetic analyses were conducted using a qualitative real-time PCR. RESULTS: The presence of B. cereus sensu lato was confirmed in 130 out of the 192 samples of convenience foods, at contamination levels ranging from 1.65 to 3.32 log CFU/g. Among the strains confirmed to belong to the B. cereus group, 23 were identified as emetic B. cereus. The analysis of each strain's ability to grow at temp. 4-10°C demonstrated that 4.9% and 12.7% of the isolates were able to grow at 4°C and 6°C, respectively. In turn, 15.2% were able to grow at 8°C, and 36.3% at 10°C. None of the psychrotrophic strains possessed genes typical of B. weihenstephanensis. The group of psychrotrophic B. cereus included potentially toxigenic strains being carriers of genes that determine the synthesis of the following toxins: NHE, HBL, CytK, and cereulide. Some of them were potent enough to produce more than one toxin. CONCLUSIONS: The analyses conducted in this study demonstrate that the psychrotolerant strains of B. cereus (including the toxigenic ones) are frequent microbiological contaminants of RTEF products offered in retail. The presence of emetic strains from the B. cereus group, which are able to grow in a wide range of temperatures and produce enterotoxins and enzymes with the characteristics of toxins, in ready-to-eat foods may pose a real threat to consumer health.

Bacterial contaminants and their antibiotic susceptibility patterns in ready-to-eat foods vended in Ogun state, Nigeria.

Contamination of ready-to-eat (RTE) foods by pathogenic bacteria may predispose consumers to foodborne diseases. This study investigated the presence of bacterial contaminants and their antibiotic susceptibility patterns in three locally processed RTE foods (eko, fufu and zobo) vended in urban markets in Ogun state, Nigeria. Bacteria isolated from a total of 120 RTE food samples were identified by 16S rRNA gene phylogeny while susceptibility patterns to eight classes of antibiotics were determined by the disc diffusion method. Species belonging to the genera Acinetobacter and Enterobacter were recovered from all RTE food types investigated, Klebsiella and Staphylococcus were recovered from eko and fufu samples, while those of Shigella were recovered from eko samples. Enterobacter hormaechei was the most prevalent species in all three RTE food types. Precisely 99% of 149 isolates were multidrug-resistant, suggesting a high risk for RTE food handlers and consumers. Co-resistance to ampicillin and cephalothin was the most frequently observed resistance phenotype. Results demonstrate that improved hygiene practices by food processors and vendors are urgently required during RTE processing and retail. Also, adequate food safety guidelines, regulation and enforcement by relevant government agencies are needed to improve the safety of RTE foods and ensure the protection of consumer health.

Microbiological safety of ready-to-eat foods in low- and middle-income countries: A comprehensive 10-year (2009 to 2018) review.

Ready-to-eat foods (RTEs) are foods consumed without any further processing. They are widely consumed as choice meals especially by school-aged children and the fast-paced working class in most low- and middle-income countries (LMICs), where they contribute substantially to the dietary intake. Depending on the type of processing and packaging material, RTEs could be industrially or traditionally processed. Typically, RTE vendors are of low literacy level, as such, they lack knowledge about good hygiene and food handling practices. In addition, RTEs are often vended in outdoor environments such that they are exposed to several contaminants of microbial origin. Depending on the quantity and type of food contaminant, consumption of contaminated RTEs may result in foodborne diseases and several other adverse health effects in humans. This could constitute major hurdles to growth and development in LMICs. Therefore, this review focuses on providing comprehensive and recent occurrence and impact data on the frequently encountered contaminants of microbial origin published in LMICs within the last decade (2009 to 2018). We have also suggested viable food safety solutions for preventing and controlling the food contamination and promoting consumer health.

Listeria monocytogenes environmental sampling program in ready-to-eat processing facilities: A practical approach.

Listeria monocytogenes is a foodborne pathogen that is frequently found in the environment. It can easily enter food processing environments and contaminate food, potentially causing public health issues. Food business operators (FBOs) are responsible for the control of L. monocytogenes in the food processing environment, particularly in facilities producing ready-to-eat food. The design and implementation of an effective environmental monitoring program (EMP) for L. monocytogenes is an integral part of controlling L. monocytogenes. An effective EMP, including all aspects from sampling, to analysis, to data interpretation, to implementation of corrective actions (including food disposition), is a tool that will help with identification and control of L. monocytogenes contamination. It should be used in conjunction with end product testing, not as a replacement for it. An EMP should be specifically designed for a particular facility on a case-by-case risk-based approach, by a food safety team within the facility. It should be reviewed regularly (at least every 6 months) and verified for its effectiveness. The control of L. monocytogenes in the food industry involves the full commitment of management and of all personnel involved with the safety of foods placed on the market, thus reducing the risk of listeriosis to consumers. Several regulatory and guidance documents provide recommendations for designing aspects of an effective L. monocytogenes EMP. However, a comprehensive review of the key components of an EMP in a single document is lacking. The objective of the present review is to provide FBOs with a practical guide to design, implementation, and verification of an EMP tailored by the food safety team for each food business.

Prevalence of ß-Lactam Drug-Resistance Genes in Escherichia coli Contaminating Ready-to-Eat Lettuce.

Thirty-four Escherichia coli isolates from 91 ready-to-eat lettuce packages, obtained from local supermarkets in Northern California, were genotyped by multilocus sequence typing, tested for susceptibility to antimicrobial agents, and screened for ß-lactamase genes. We found 15 distinct sequence types (STs). Six of these genotypes (ST1198, ST2625, ST2432, ST2819, ST4600, and ST5143) have been reported as pathogens found in human samples. Twenty-six (76%) E. coli isolates were resistant to ampicillin, 17 (50%) to ampicillin/sulbactam, 8 (23%) to cefoxitin, and 7 (20%) to cefuroxime. blaCTX-M was the most prevalent ß-lactamase gene, identified in eight (23%) isolates. We identified a class A broad-spectrum ß-lactamase SED-1 gene, blaSED, reported by others in Citrobacter sedlakii isolated from bile of a patient. This study found that fresh lettuce carries ß-lactam drug-resistant E. coli, which might serve as a reservoir for drug-resistance genes that could potentially be transmitted to pathogens that cause human infections.

Metagenomic characterization of bacterial community and antibiotic resistance genes in representative ready-to-eat food in southern China.

Ready-to-eat (RTE) foods have been considered to be reservoirs of antibiotic resistance bacteria, which constitute direct threat to human health, but the potential microbiological risks of RTE foods remain largely unexplored. In this study, the metagenomic approach was employed to characterize the comprehensive profiles of bacterial community and antibiotic resistance gene (ARG) in 18 RTE food samples (8 RTE meat, 7 RTE vegetables and 3 RTE fruit) in southern China. In total, the most abundant phyla in RTE foods were Proteobacteria, Firmicutes, Cyanobacteria, Bacteroidetes and Actinobacteria. 204 ARG subtypes belonging to 18 ARG types were detected with an abundance range between 2.81 × 10-5 and 7.7 × 10-1 copy of ARG per copy of 16S rRNA gene. Multidrug-resistant genes were the most predominant ARG type in the RTE foods. Chloramphenicol, macrolide-lincosamide-streptogramin, multidrug resistance, aminoglycoside, bacitracin, tetracycline and ß-lactam resistance genes were dominant, which were also associated with antibiotics used extensively in human medicine or veterinary medicine/promoters. Variation partitioning analysis indicated that the join effect of bacterial community and mobile genetic elements (MGEs) played an important role in the resistome alteration. This study further deepens the comprehensive understanding of antibiotic resistome and the correlations among the antibiotic resistome, microbiota, and MGEs in the RTE foods.

Microbiological safety of ready-to-eat fresh-cut fruits and vegetables sold on the Canadian retail market.

Following implementation of Health Canada's Policy on Listeria monocytogenes in Ready-to-Eat Foods by Canadian food safety authorities in 2011, a four-year study (2012-2016) was carried out to gain baseline information on the occurrence of bacterial pathogens, notably the prevalence and levels of Listeria monocytogenes (L. monocytogenes) in various product types of ready-to-eat (RTE) fresh-cut fruits and fresh-cut vegetables sold at retail in Canada. A total of 10,070 pre-packaged samples, including 4691 fresh-cut fruit and 5379 fresh-cut vegetable samples were collected from retail stores across Canada and analyzed for bacterial pathogens and generic Escherichia coli (E. coli). Salmonella species (spp.), E. coli O157:H7, Shigella and Campylobacter were not detected in any of the tested samples. L. monocytogenes was identified in 0.51% (95% CI [0.34, 0.76]) of the fresh-cut fruit and in 0.24% (95% CI [0.14, 0.41]) of the fresh-cut vegetable samples. Of the 37 L. monocytogenes positive samples identified, levels of L. monocytogenes <5 CFU/g, 5-<100 CFU/g, and ≥100 CFU/g were found in 67.6% (25/37), 24.3% (9/37) and 8.1% (3/37) of the samples, respectively. The results of this study indicate that the vast majority of fresh-cut fruits and vegetables sold on the Canadian retail market are safe for consumption. However, contamination by L. monocytogenes can infrequently occur in fresh-cut fruits and vegetables, with certain types of fresh-cut fruits (i.e., melons, apples) and vegetables (i.e., mushrooms, cauliflower) being more likely to be contaminated than others. Safe handling practices are recommended for producers, retailers and consumers including storage at refrigerated temperatures.