Phylogenetic identification of bacterial MazF toxin protein motifs among probiotic strains and foodborne pathogens and potential implications of engineered probiotic intervention in food

BACKGROUND: Toxin-antitoxin (TA) systems are commonly found in bacteria and Archaea, and it is the most common mechanism involved in bacterial programmed cell death or apoptosis. Recently, MazF, the toxin component of the toxin-antitoxin module, has been categorized as an endoribonuclease, or it may have a function similar to that of a RNA interference enzyme. RESULTS: In this paper, with comparative data and phylogenetic analyses, we are able to identify several potential MazF-conserved motifs in limited subsets of foodborne pathogens and probiotic strains and further provide a molecular basis for the development of engineered/synthetic probiotic strains for the mitigation of foodborne illnesses. Our findings also show that some probiotic strains, as fit as many bacterial foodborne pathogens, can be genetically categorized into three major groups based on phylogenetic analysis of MazF. In each group, potential functional motifs are conserved in phylogenetically distant species, including foodborne pathogens and probiotic strains. CONCLUSION: These data provide important knowledge for the identification and computational prediction of functional motifs related to programmed cell death. Potential implications of these findings include the use of engineered probiotic interventions in food or use of a natural probiotic cocktail with specificity for controlling targeted foodborne pathogens

Salmonella - Distribution, Adaptation, Control Measures and Molecular Technologies

The discovery of Salmonella in swine in 1885 marked the beginning of intense efforts to control salmonellae that have continued for the past 127 years. The majority of foodborne outbreaks are caused by only a few of the 2500+ known serovars. While progress has been made on many fronts, salmonellosis has yet to be eliminated in either developed or in developing nations. This work represents the collective contributions of authors from all around the world. Chapters in this book address a wide array of topics related to understanding and controlling this pathogen, including: Salmonella as studied in the environment, air and in food products; virulence and pathogenicity; control by bacteriophages and other antimicrobials; bacterial adaptation; etc

Possible Sources of Listeria monocytogenes Contamination of Fresh-cut Apples and Antimicrobial Interventions During Antibrowning Treatments: A Review

Fresh-cut apples, being rich in antioxidants and other nutrients, have emerged as popular snacks in restaurants, at home, and in school lunch programs, partially due to freshness, convenience, and portion size availability. Two major challenges in processing fresh-cut apples are the browning of cut surfaces and contamination with human pathogens. Regarding human pathogens, contamination by Listeria monocytogenes is a major concern, as evidenced by two outbreaks of whole apples and numerous recalls of fresh-cut apples. Antibrowning agents currently used by the industry have little to no antimicrobial properties. The present review discusses the possible origins of L. monocytogenes in fresh-cut apples, including contaminated whole apples, and contamination via the processing environment and the equipment in fresh-cut facilities. Treatment with antibrowning solutions could possibly be an opportunity for Listeria contamination and represents the last chance to inactivate pathogens. The discussion is focused on the antibrowning treatments where formulations and coatings with antibrowning and antimicrobial properties have been developed and evaluated against Listeria and other microorganisms. In addition, several research needs and considerations are discussed to further reduce the chance of pathogen contamination on fresh-cut apples

Minimum Concentrations of Slow Pyrolysis Paper and Walnut Hull Cyclone Biochars Needed to Inactivate Escherichia coli O157:H7 in Soil

Antimicrobial properties of biochar have been attributed to its ability to inactivate foodborne pathogens in soil, to varying degrees. High concentrations of biochar have reduced E. coli O157:H7 in soil and dairy manure compost, based on alkaline pH. Preliminary studies evaluating 31 different biochars determined that two slow pyrolysis biochars (paper biochar and walnut hull cyclone biochar) were the most effective at inactivating E. coli in soil. A study was conducted to determine the lowest percentages of paper and walnut hull cyclone biochars needed to reduce E. coli O157:H7 in soil. A model soil was adjusted to 17.75% moisture, and the two types of biochar were added at concentrations of 1.0, 1.5, 2.0, 2.5, 3.5, 4.5, 5.5, and 6.5%. Nontoxigenic E. coli O157:H7 were inoculated into soil at 6.84 log CFU/g and stored for up to 6 weeks at 21°C. Mean E. coli O157:H7 counts were 6.01–6.86 log CFU/g at all weeks between 1 and 6 in soil-only positive control samples. Populations in all soil amended with 1.0 and 1.5% of either type of biochar (as well as 2.0% of the walnut hull biochar) resulted in ≤0.68 log reductions at week 6, when compared with positive controls. All other concentrations (i.e., ≥2.0% paper and ≥2.5% walnut hull) inactivated ≥2.7 log at all weeks between 1 and 6 (p < 0.05). At the end of 6 weeks, E. coli O157:H7 declined by 2.84 log in 2.0% paper biochar samples, while concentrations of between 2.5 and 6.5% paper biochar completely inactivated E. coli O157:H7, as determined by spiral plating, at weeks 5 and 6. In contrast, 2.0% walnut hull biochar lowered populations by only 0.38 log at week 6, although 2.5–6.5% concentrations of walnut hull biochar resulted in complete inactivation at all weeks between 3 and 6, as assessed by spiral plating. In summary, ≥2.5% paper or walnut hull biochar reduced ≥5.0 log of E. coli O157:H7 during the 6-week storage period, which we attribute to high soil alkalinity. Amended at a 2.5% concentration, the pH of soil with paper or walnut hull biochar was 10.67 and 10.06, respectively. Results from this study may assist growers in the use of alkaline biochar for inactivating E. coli O157:H7 in soil

Comprehensive Approaches to Molecular Biomarker Discovery for Detection and Identification of Cronobacter spp. (Enterobacter sakazakii) and Salmonella spp. ▿

Cronobacter spp. (formerly Enterobacter sakazakii) and Salmonella spp. are increasingly implicated internationally as important microbiological contaminants in low-moisture food products, including powdered infant formula. Estimates indicate that 40 to 80% of infants infected with Cronobacter sakazakii and/or Salmonella in the United States may not survive the illness. A systematic approach, combining literature-based data mining, comparative genome analysis, and the direct sequencing of PCR products of specific biomarker genes, was used to construct an initial collection of genes to be targeted. These targeted genes, particularly genes encoding virulence factors and genes responsible for unique phenotypes, have the potential to function as biomarker genes for the identification and differentiation of Cronobacter spp. and Salmonella from other food-borne pathogens in low-moisture food products. In this paper, a total of 58 unique Salmonella gene clusters and 126 unique potential Cronobacter biomarkers and putative virulence factors were identified. A chitinase gene, a well-studied virulence factor in fungi, plants, and bacteria, was used to confirm this approach. We found that the chitinase gene has very low sequence variability and/or polymorphism among Cronobacter, Citrobacter, and Salmonella, while differing significantly in other food-borne pathogens, either by sequence blasting or experimental testing, including PCR amplification and direct sequencing. This computational analysis for Cronobacter and Salmonella biomarker identification and the preliminary laboratory studies are only a starting point; thus, PCR and array-based biomarker verification studies of these and other food-borne pathogens are currently being conducted