The Analysis of Field Strains Isolated From Food, Animal and Clinical Sources Uncovers Natural Mutations in Listeria monocytogenes Nisin Resistance Genes.

Nisin is a commonly used bacteriocin for controlling spoilage and pathogenic bacteria in food products. Strains possessing high natural nisin resistance that reduce or increase the potency of this bacteriocin against Listeria monocytogenes have been described. Our study sought to gather more insights into nisin resistance mechanisms in natural L. monocytogenes populations by examining a collection of 356 field strains that were isolated from different foods, food production environments, animals and human infections. A growth curve analysis-based approach was used to access nisin inhibition levels and assign the L. monocytogenes strains into three nisin response phenotypic categories; resistant (66%), intermediate (26%), and sensitive (8%). Using this categorization isolation source, serotype, genetic lineage, clonal complex (CC) and strain-dependent natural variation in nisin phenotypic resistance among L. monocytogenes field strains was revealed. Whole genome sequence analysis and comparison of high nisin resistant and sensitive strains led to the identification of new naturally occurring mutations in nisin response genes associated with increased nisin resistance and sensitivity in this bacterium. Increased nisin resistance was detected in strains harboring RsbUG77S and PBPB3V240F amino acid substitution mutations, which also showed increased detergent stress resistance as well as increased virulence in a zebra fish infection model. On the other hand, increased natural nisin sensitivity was detected among strains with mutations in sigB, vir, and dlt operons that also showed increased lysozyme sensitivity and lower virulence. Overall, our study identified naturally selected mutations involving pbpB3 (lm0441) as well as sigB, vir, and dlt operon genes that are associated with intrinsic nisin resistance in L. monocytogenes field strains recovered from various food and human associated sources. Finally, we show that combining growth parameter-based phenotypic analysis and genome sequencing is an effective approach that can be useful for the identification of novel nisin response associated genetic variants among L. monocytogenes field strains.

Quantitative Risk Assessment of Hemolytic Uremic Syndrome Associated with Consumption of Bulk Milk Sold Directly from Producer to Consumer in South Africa.

This study was conducted to estimate the hemolytic uremic syndrome (HUS) risk associated with consumption of producer-distributor bulk milk (PDBM) contaminated with Shiga toxin-producing Escherichia coli (STEC) in South Africa. Data were obtained from recently completed studies in South Africa taking into account prior collected prevalence data of STEC in raw and pasteurized PDBM and survey information from producer-distributor outlets and households. Inputs for the models were complemented with data from published and unpublished literature. A probabilistic exposure model was developed with Monte Carlo simulation in Excel add-in software using @Risk software. Hazard characterization was based on an exponential dose-response model to calculate the probability of illness from STEC infection in individuals 5 years and younger and individuals older than 5 years. The estimated mean STEC level was 0.12 CFU/mL (95% confidence interval [CI]: 0 to 1.2; σ = 0.34) for raw PDBM and 0.08 CFU/mL (95% CI: 0 to 1; σ = 0.27) for pasteurized PDBM. A higher risk of HUS cases per year was recorded in raw than in pasteurized PDBM and also in individuals younger than 5 years of age. For every 100,000 servings consumed, the expected median numbers of HUS cases per year from raw PDBM were 52 for 5 years and younger and 3.2 for older than 5 years. The median numbers of cases per year for pasteurized PDBM were 47 for 5 years and younger and 2.9 for older than 5 years. Sensitivity analysis revealed that serving volume and time taken to sell PDBM at producer-distributor outlets were the factors with the greatest impact on probability of illness. The models developed in this study are an example of risk assessments for milk produced and marketed from similar scenarios across the globe.

The Impact of Cooling Rate on the Safety of Food Products as Affected by Food Containers.

In recent decades, the demand for ready-to-eat (RTE) food items prepared by the food catering sector has increased together with the value of cook-serve, cook-chill, and cook-freeze food products. The technologies by which foods are cooked, chilled, refrigerated for storage, and reheated before serving are of prime importance to maintain safety. Packaging materials and food containers play an important role in influencing the cooling rate of RTE foods. Food items that are prepared using improper technologies and inappropriate packaging materials may be contaminated with foodborne pathogens. Numerous research studies have shown the impact of deficient cooling technologies on the survival and growth of foodborne pathogens, which may subsequently pose a threat to public health. The operating temperatures and cooling rates of the cooling techniques applied must be appropriate to inhibit the growth of pathogens. Food items must be stored outside the temperature danger zone, which is between 5 and 60 °C, in order to inhibit the growth of these pathogens. The cooling techniques used to prepare potentially hazardous foods, such as cooked meat, rice, and pasta, must be properly applied and controlled to ensure food safety. This paper critically reviews the effects of cooling and its relationship to food containers on the safety of RTE foods produced and sold through the food service industry.

Application of Probabilistic Modeling to Quantify the Reduction Levels of Hepatocellular Carcinoma Risk Attributable to Chronic Aflatoxins Exposure.

Epidemiological studies show a definite connection between areas of high aflatoxin content and a high occurrence of human hepatocellular carcinoma (HCC). Hepatitis B virus in individuals further increases the risk of HCC. The two risk factors are prevalent in rural Kenya and continuously predispose the rural populations to HCC. A quantitative cancer risk assessment therefore quantified the levels at which potential pre- and postharvest interventions reduce the HCC risk attributable to consumption of contaminated maize and groundnuts. The assessment applied a probabilistic model to derive probability distributions of HCC cases and percentage reductions levels of the risk from secondary data. Contaminated maize and groundnuts contributed to 1,847 ± 514 and 158 ± 52 HCC cases per annum, respectively. The total contribution of both foods to the risk was additive as it resulted in 2,000 ± 518 cases per annum. Consumption and contamination levels contributed significantly to the risk whereby lower age groups were most affected. Nonetheless, pre- and postharvest interventions might reduce the risk by 23.0-83.4% and 4.8-95.1%, respectively. Therefore, chronic exposure to aflatoxins increases the HCC risk in rural Kenya, but a significant reduction of the risk can be achieved by applying specific pre- and postharvest interventions.

Pathogenic and commensal Escherichia coli from irrigation water show potential in transmission of extended spectrum and AmpC ß-lactamases determinants to isolates from lettuce.

There are few studies on the presence of extended-spectrum ß-lactamases and AmpC ß-lactamases (ESBL/AmpC) in bacteria that contaminate vegetables. The role of the production environment in ESBL/AmpC gene transmission is poorly understood. The occurrence of ESBL/AmpC in Escherichia coli (n = 46) from lettuce and irrigation water and the role of irrigation water in the transmission of resistant E. coli were studied. The presence of ESBL/AmpC, genetic similarity and phylogeny were typed using genotypic and phenotypic techniques. The frequency of ß-lactamase gene transfer was studied in vitro. ESBLs/AmpC were detected in 35 isolates (76%). Fourteen isolates (30%) produced both ESBLs/AmpC. Prevalence was highest in E. coli from lettuce (90%). Twenty-two isolates (48%) were multi-resistant with between two and five ESBL/AmpC genes. The major ESBL determinant was the CTX-M type (34 isolates). DHA (33% of isolates) were the dominant AmpC ß lactamases. There was a high conjugation efficiency among the isolates, ranging from 3.5 × 10(-2) to 1 × 10(-2) ± 1.4 × 10(-1) transconjugants per recipient. Water isolates showed a significantly higher conjugation frequency than those from lettuce. A high degree of genetic relatedness between E. coli from irrigation water and lettuce indicated possible common ancestry and pathway of transmission.

Lactic acid bacteria and yeasts involved in the fermentation ofamabere amaruranu, a Kenyan fermented milk.

Indigenous fermented milk products contain microbiota composed of technologically important species and strains which are gradually getting lost with new technologies. We investigated the microbial diversity inamabere amaruranu, a traditionally fermented milk product from Kenya. Sixteen samples of the product from different containers were obtained. One hundred and twenty isolates of lactic acid bacteria (LAB) and 67 strains of yeasts were identified using API 50 CH and API 20 C AUX identification kits, respectively. The average pH of all the traditional fermented samples was 4.00 ± 0.93. Lactobacilli, yeasts, and molds as well asEnterobacteriaceae counts from the plastic containers were significantly higher (P < 0.05) than those from gourd.Enterobacteriaceae were below 1.00 ± 1.11 log10 cfu/mL in products from the gourds and 2.17 ± 1.92 log10 cfu/mL from the plastic containers. The LAB species were identified asStreptococcus thermophilus (25%),Lactobacillus plantarum (20%), andLeuconostoc mesenteroides (20%). The predominant yeasts wereSaccharomyces cerevisiae (25%),Trichosporum mucoides (15%),Candida famata (10%), andCandida albicans (10%). The type of vessel used for fermentation had no significant influence on the type of isolated and identified species. The diverse mixture of LAB and yeasts microflora forms a potential consortium for further product innovation inamabere amaruranu and other fermented milk products.