Genomic and Phenotypic Analysis of Bacteriophages Identifies Two Novel Phage Species.

Bacteriophages (phages) are potential alternatives to chemical antimicrobials against pathogens of public health significance. Understanding the diversity and host specificity of phages is important for developing effective phage biocontrol approaches. Here, we assessed the host range, morphology, and genetic diversity of eight phages isolated from a wastewater treatment plant.

How Broad Is Enough: The Host Range of Bacteriophages and Its Impact on the Agri-Food Sector.

Novel bacteriophages (phages) possessing a broad host range are consistently and routinely reported, yet there is presently no consensus on the definition of "broad host range." As phages are increasingly being used in the development of methods for the detection and biocontrol of human pathogens, it is important to address the limitations associated with the host range. For instance, unanticipated host range breadth may result in the detection of nonpathogenic targets, thereby increasing the false-positive rate.

Antibiotic Resistance in Shiga Toxigenic Isolates from Surface Waters and Sediments in a Mixed Use Urban Agricultural Landscape.

Antibiotic resistance (AR) phenotypes and acquired resistance determinants (ARDs) detected by in silico analysis of genome sequences were examined in 55 Shiga toxin-producing (STEC) isolates representing diverse serotypes recovered from surfaces waters and sediments in a mixed use urban/agricultural landscape in British Columbia, Canada. The isolates displayed decreased susceptibility to florfenicol (65.5%), chloramphenicol (7.

Endogenous Metabolites Released by Sanitized Sprouting Alfalfa Seed Inhibit the Growth of Salmonella enterica.

Sprouts are the leading cause of foodborne disease outbreaks globally, mainly because the specialized conditions required to germinate seed sprouts for human consumption contribute to an environment that allows pathogenic bacteria to flourish. To reduce risk of illness, current food safety guidelines in the United States and Canada recommend hypochlorite treatment for seed sanitation. However, many growers and consumers have become wary of the impact of hypochlorite on human health and the environment and are actively seeking less caustic approaches.

Natural Compounds With Antibacterial Activity Against spp. in Powdered Infant Formula: A Review.

Bacteria from the genus are opportunistic foodborne pathogens capable of causing severe infections in neonates, the elderly and immunocompromised adults. The majority of neonatal infections have been linked epidemiologically to dehydrated powdered infant formulas (PIFs), the majority of which are manufactured using processes that do not ensure commercial sterility. Unfortunately, the osmotolerance, desiccation resistance, mild thermotolerance and wide-ranging minimum, optimum and maximum growth temperatures of spp.

Inactivation of on post-harvest cantaloupe and lettuce by a lytic bacteriophage cocktail.

() is a causative agent of multiple outbreaks of foodborne illness associated with fresh produce, including pre-cut melon and leafy vegetables. Current industrial antimicrobial interventions have been shown to reduce microbial populations by <90%. Consequently, bacteriophages have been suggested as an alternative to chemical sanitizers.

Bacteriophage-Insensitive Mutants of Antimicrobial-Resistant are Altered in their Tetracycline Resistance and Virulence in Caco-2 Intestinal Cells.

Bacteriophages have shown promise as therapeutic alternatives to antibiotics for the control of infectious bacteria, including the human pathogen However, the development of effective phage-based applications requires the elucidation of key interactions between phages and target hosts, particularly since host resistance to phage is inevitable. Little is known about the alteration of host phenotypes following the development of resistance to phage. The aim of this study is to evaluate the antibiotic susceptibility and virulence of a isolate following the development of resistance to bacteriophage SI1.

Viable but Nonculturable Escherichia coli O157:H7 and Salmonella enterica in Fresh Produce: Rapid Determination by Loop-Mediated Isothermal Amplification Coupled with a Propidium Monoazide Treatment.

O157:H7 and are leading causes of foodborne outbreaks linked to fresh produce. Both species can enter the "viable but nonculturable" (VBNC) state that precludes detection using conventional culture-based or molecular methods. In this study, we assessed propidium monoazide-quantitative PCR (PMA-qPCR) assays and novel methods combining PMA and loop-mediated isothermal amplification (LAMP) for the detection and quantification of VBNC O157:H7 and in fresh produce.

Disinfection of Alfalfa and Radish Sprouting Seed Using Oxidizing Agents and Treatments Compliant with Organic Food Production Principles.

Abstract: Antimicrobial seed treatments recommended by Canadian guidance for sprouted vegetable production (2,000 ppm of hypochlorite for 15 to 20 min or 6 to 10% hydrogen peroxide for 10 min at room temperature) are not fully compliant with organic production principles. We investigated the effect of a sequential treatment consisting of a 10-min soak at 50°C in water followed by exposure to a 2.0% H2O2 plus 0.

Reaction of Surrogate Serotype O157:H7 and Non-O157 Strains to Nutrient Starvation: Variation in Phenotype and Transcription of Stress Response Genes and Behavior on Lettuce Plants in the Field.

Preharvest contamination with bacteria borne by irrigation water may result in leafy vegetables serving as vehicles for transmission of Shiga toxin-producing (STEC) to humans. The influence of starvation-associated stress on the behavior of non-toxin-producing strains of serotype O157:H7 and serotypes O26, O103, O111, and O145 was examined subsequent to their introduction to the phyllosphere of field-grown romaine lettuce as inocula simulating starved (96 h in sterile deionized water) and nutrient-depleted (24 h broth culture) cells. As with O157:H7, leaf populations of the non-O157 strains declined rapidly during the first 72 h postinoculation, displaying the biphasic decay curve typical of serotype O157:H7 isolates.

Diversity and Host Specificity Revealed by Biological Characterization and Whole Genome Sequencing of Bacteriophages Infecting .

Phages infecting members of the opportunistic human pathogen, , are widespread in natural environments and offer a potential source of agents that could be used for controlling populations of this bacterium; yet, relatively little is known about these phages. Here we describe the isolation and characterization of 45 phages of from disparate geographic locations within British Columbia, Canada. Host-range profiling revealed host-specific patterns of susceptibility and resistance, with several phages identified that have a broad-host range (i.

Effects of wild blueberry (Vaccinium angustifolium) pomace feeding on gut microbiota and blood metabolites in free-range pastured broiler chickens.

There is a need to develop cost-effective approaches to modulate gut microbiota, promote bird health, and prevent infections in pasture-raised broiler chickens. The present study evaluated the efficacy of organic wild blueberry (Vaccinium angustifolium) also called low-bush blueberry pomace (LBBP)-supplemented feed to modulate the chicken gut microbiota, and blood metabolites in order to improve bird health and productivity. Slow-growing broiler chickens were reared on pasture up to 64 D for sampling after 2 wk of treatment during brooding with 0, 1, and 2% LBBP in feed.

Pathogen reduction on mung bean reduction of Escherichia coli O157:H7, Salmonella enterica and Listeria monocytogenes on mung bean using combined thermal and chemical treatments with acetic acid and hydrogen peroxide.

Mung bean sprouts were implicated in several foodborne outbreaks worldwide in recent years. The objectives of this study were to (i) assess the efficacy of individual (mild heat) and combined treatments (mild heat followed by acetic acid or/and hydrogen peroxide) for the inactivation of enteric bacterial pathogens on mung bean intended for sprout production and (ii) determine the impact of the treatments and storage conditions on germination. Mung bean was co-inoculated with Escherichia coli O157:H7, Salmonella enterica and Listeria monocytogenes to achieve initial populations of approximately 5-6 log CFU of each species/g bean.

Characterization of Four Novel Bacteriophages Isolated from British Columbia for Control of Non-typhoidal and on Sprouting Alfalfa Seeds.

Alfalfa sprouts have been linked to numerous North American outbreaks of in recent years. Conventionally, treatments involving chlorine, heat, and irradiation are used for alfalfa seed sanitation. However, such treatments may be highly variable in their efficacy for pathogen control and/or detrimental to sprout quality, therefore negatively perceived by consumers advocating for natural alternatives.

A Syst-OMICS Approach to Ensuring Food Safety and Reducing the Economic Burden of Salmonellosis.

The Syst-OMICS consortium is sequencing 4,500 genomes and building an analysis pipeline for the study of genome evolution, antibiotic resistance and virulence genes. Metadata, including phenotypic as well as genomic data, for isolates of the collection are provided through the Foodborne Syst-OMICS database (SalFoS), at https://salfos.ibis.

Phenotypic and Genotypic Characteristics of Shiga Toxin-Producing Escherichia coli Isolated from Surface Waters and Sediments in a Canadian Urban-Agricultural Landscape.

A hydrophobic grid membrane filtration-Shiga toxin immunoblot method was used to examine the prevalence of Shiga toxin-producing Escherichia coli (STEC) in four watersheds located in the Lower Mainland of British Columbia, Canada, a region characterized by rapid urbanization and intensive agricultural activity. STEC were recovered from 21.6, 23.

Fate of Salmonella enterica in a mixed ingredient salad containing lettuce, cheddar cheese, and cooked chicken meat.

Food service and retail sectors offer consumers a variety of mixed ingredient salads that contain fresh-cut vegetables and other ingredients such as fruits, nuts, cereals, dairy products, cooked seafood, cooked meat, cured meats, or dairy products obtained from external suppliers. Little is known about the behavior of enteric bacterial pathogens in mixed ingredient salads. A model system was developed to examine the fate of Salmonella enterica (inoculum consisting of S.

Microbiological survey of locally grown lettuce sold at farmers' markets in Vancouver, British Columbia.

Increased consumer demand for fresh leafy produce has been paralleled by an increase in outbreaks and illness associated with these foods. Presently, data on the microbiological quality and safety of produce harvested in the Lower Mainland of British Columbia is lacking. Therefore, fresh green, red, and romaine lettuce samples (n = 68) were obtained from five regional farmers' markets in late summer of 2012 and subsequently analyzed to determine total numbers of aerobic bacteria, coliforms, and Escherichia coli.

Exploiting the explosion of information associated with whole genome sequencing to tackle Shiga toxin-producing Escherichia coli (STEC) in global food production systems.

The rates of foodborne disease caused by gastrointestinal pathogens continue to be a concern in both the developed and developing worlds. The growing world population, the increasing complexity of agri-food networks and the wide range of foods now associated with STEC are potential drivers for increased risk of human disease. It is vital that new developments in technology, such as whole genome sequencing (WGS), are effectively utilized to help address the issues associated with these pathogenic microorganisms.

Evaluation of different approaches for modeling Escherichia coli O157:H7 survival on field lettuce.

The ability to predict the behavior of Escherichia coli O157:H7 on contaminated field lettuce is essential for the development of accurate quantitative microbial risk assessments. The survival pattern of the species was assessed from several data sets derived from field-based experiments, which were analyzed by regression analysis fitting one monophasic model (log-linear) and two biphasic (Weibull and Cerf's model) models. Probabilistic models were also simulated with @RISK™, integrating the fitted monophasic and biphasic models in order to analyze their impact on the estimate of the extent of die-off subsequent to a contamination event in the field.

Duplex PCR methods for the molecular detection of Escherichia fergusonii isolates from broiler chickens.

Escherichia fergusonii is an emerging pathogen that has been isolated from a wide range of infections in animals and humans. Primers targeting specific genes, including yliE (encoding a conserved hypothetical protein of the cellulose synthase and regulator of cellulose synthase island), EFER_1569 (encoding a hypothetical protein, putative transcriptional activator for multiple antibiotic resistance), and EFER_3126 (encoding a putative triphosphoribosyl-dephospho-coenzyme A [CoA]), were designed for the detection of E. fergusonii by conventional and real-time PCR methods.

Antibiotic resistance and diversity of Salmonella enterica serovars associated with broiler chickens.

The objective of this study was to analyze the antibiotic resistance phenotype and genotype of Salmonella isolated from broiler production facilities. A total of 193 Salmonella isolates recovered from commercial farms in British Columbia, Canada, were evaluated. Susceptibility to antibiotics was determined with the Sensititre system.

Optimized extraction and characterization of antimicrobial phenolic compounds from mangosteen (Garcinia mangostana L.) cultivation and processing waste.

Background: Applications for antimicrobials derived from the mangosteen (Garcinia mangostana L.) plant are presently restricted by high production costs. Extraction from cultivation or processing waste streams using a solvent-free approach could lessen to permit commercial applications in food processing and preservation.

A framework for developing research protocols for evaluation of microbial hazards and controls during production that pertain to the quality of agricultural water contacting fresh produce that may be consumed raw.

Agricultural water may contact fresh produce during irrigation and/or when crop protection sprays (e.g., cooling to prevent sunburn, frost protection, and agrochemical mixtures) are applied.