@article{jayeola_farber_kathariou_2022, title={Induction of the Viable-but-Nonculturable State in Salmonella Contaminating Dried Fruit}, volume={88}, ISSN={["1098-5336"]}, DOI={10.1128/AEM.01733-21}, abstractNote={ Salmonella is a leading foodborne pathogen globally causing numerous outbreaks of foodborne illnesses and remains the leading contributor to deaths attributed to foodborne disease in the United States and other industrialized nations. Therefore, efficient detection methods for Salmonella contaminating food are critical for public health and food safety. }, number={2}, journal={APPLIED AND ENVIRONMENTAL MICROBIOLOGY}, author={Jayeola, Victor and Farber, J. M. and Kathariou, S.}, year={2022}, month={Jan} } @article{jayeola_mcclelland_porwollik_chu_farber_kathariou_2020, title={Identification of Novel Genes Mediating Survival ofSalmonellaon Low-Moisture Foods via Transposon Sequencing Analysis}, volume={11}, ISSN={["1664-302X"]}, DOI={10.3389/fmicb.2020.00726}, abstractNote={Salmonella enterica is the leading foodborne pathogen associated with outbreaks involving low-moisture foods (LMFs). However, the genes involved in Salmonella’s long-term survival on LMFs remain poorly characterized. In this study, in-shell pistachios were inoculated with Tn5-based mutant libraries of S. Enteritidis P125109, S. Typhimurium 14028s, and S. Newport C4.2 at approximate 108 CFU/g and stored at 25°C. Transposon sequencing analysis (Tn-seq) was then employed to determine the relative abundance of each Tn5 insertion site immediately after inoculation (T0), after drying (T1), and at 120 days (T120). In S. Enteritidis, S. Typhimurium, and S. Newport mutant libraries, the relative abundance of 51, 80, and 101 Tn5 insertion sites, respectively, was significantly lower at T1 compared to T0, while in libraries of S. Enteritidis and S. Typhimurium the relative abundance of 42 and 68 Tn5 insertion sites, respectively, was significantly lower at T120 compared to T1. Tn5 insertion sites with reduced relative abundance in this competition assay were localized in DNA repair, lipopolysaccharide biosynthesis and stringent response genes. Twelve genes among those under strong negative selection in the competition assay were selected for further study. Whole gene deletion mutants in ten of these genes, sspA, barA, uvrB, damX, rfbD, uvrY, lrhA, yifE, rbsR, and ompR, were impaired for individual survival on pistachios. The findings highlight the value of combined mutagenesis and sequencing to identify novel genes important for the survival of Salmonella in low-moisture foods.}, journal={FRONTIERS IN MICROBIOLOGY}, author={Jayeola, Victor and McClelland, Michael and Porwollik, Steffen and Chu, Weiping and Farber, Jeffrey and Kathariou, Sophia}, year={2020}, month={May} } @article{jayeola_parsons_gorski_kathariou_2019, title={Validation of an ampicillin selection protocol to enrich for mutants of Listeria monocytogenes unable to replicate on fresh produce}, volume={366}, ISSN={["1574-6968"]}, DOI={10.1093/femsle/fnz076}, abstractNote={ABSTRACT Several outbreaks of listeriosis have implicated fresh produce but genetic factors required for growth of Listeria monocytogenes on produce remain poorly characterized. Based on the fact that β-lactam antibiotics only kill bacterial cells that are growing, we hypothesized that ampicillin selection can enrich for L. monocytogenes mutants unable to grow on produce. For validation, we examined relative recovery of L. monocytogenes strain 2011L-2858 and its cold-sensitive mutant L1E4 following inoculation of cantaloupe rind fragments with 1:1 mixture of the strains and incubation at 4°C with or without ampicillin. Listeria monocytogenes from rind fragments inoculated with the mixed cultures and incubated in the presence of ampicillin were used to inoculate fresh rind fragments for a second round of enrichment. In the presence of ampicillin, the proportion of L1E4 increased from 55% on day 0 to 78% on day 14, with higher recovery (85% after 14 days) in the second round of enrichment. These data suggested that L1E4 was enriched on cantaloupe rind fragments while growing cells of the wildtype were killed by ampicillin. Application of this protocol to transposon mutant libraries from three L. monocytogenes strains yielded several mutants unable to grow on cantaloupe. Thus, ampicillin selection can facilitate discovery of genes essential for growth of L. monocytogenes on fresh produce.}, number={7}, journal={FEMS MICROBIOLOGY LETTERS}, author={Jayeola, Victor and Parsons, C. and Gorski, L. and Kathariou, S.}, year={2019}, month={Apr} } @article{parsons_lee_jayeola_kathariou_2017, title={Novel Cadmium Resistance Determinant in Listeria monocytogenes}, volume={83}, ISSN={["1098-5336"]}, DOI={10.1128/aem.02580-16}, abstractNote={ABSTRACT Listeria monocytogenes is a foodborne pathogen that can cause severe disease (listeriosis) in susceptible individuals. It is ubiquitous in the environment and often exhibits resistance to heavy metals. One of the determinants that enables Listeria to tolerate exposure to cadmium is the cadAC efflux system, with CadA being a P-type ATPase. Three different cadA genes (designated cadA1 to cadA3 ) were previously characterized in L. monocytogenes . A novel putative cadmium resistance gene ( cadA4 ) was recently identified through whole-genome sequencing, but experimental confirmation for its involvement in cadmium resistance is lacking. In this study, we characterized cadA4 in L. monocytogenes strain F8027, a cadmium-resistant strain of serotype 4b. By screening a mariner-based transposon library of this strain, we identified a mutant with reduced tolerance to cadmium and that harbored a single transposon insertion in cadA4 . The tolerance to cadmium was restored by genetic complementation with the cadmium resistance cassette ( cadA4C ), and enhanced cadmium tolerance was conferred to two unrelated cadmium-sensitive strains via heterologous complementation with cadA4C . Cadmium exposure induced cadA4 expression, even at noninhibitory levels. Virulence assessments in the Galleria mellonella model suggested that a functional cadA4 suppressed virulence, potentially promoting commensal colonization of the insect larvae. Biofilm assays suggested that cadA4 inactivation reduced biofilm formation. These data not only confirm cadA4 as a novel cadmium resistance determinant in L. monocytogenes but also provide evidence for roles in virulence and biofilm formation. IMPORTANCE Listeria monocytogenes is an intracellular foodborne pathogen causing the disease listeriosis, which is responsible for numerous hospitalizations and deaths every year. Among the adaptations that enable the survival of Listeria in the environment are the abilities to persist in biofilms, grow in the cold, and tolerate toxic compounds, such as heavy metals. Here, we characterized a novel determinant that was recently identified on a larger mobile genetic island through whole-genome sequencing. This gene ( cadA4 ) was found to be responsible for cadmium detoxification and to be a divergent member of the Cad family of cadmium efflux pumps. Virulence assessments in a Galleria mellonella model suggested that cadA4 may suppress virulence. Additionally, cadA4 may be involved in the ability of Listeria to form biofilms. Beyond the role in cadmium detoxification, the involvement of cadA4 in other cellular functions potentially explains its retention and wide distribution in L. monocytogenes . }, number={5}, journal={APPLIED AND ENVIRONMENTAL MICROBIOLOGY}, author={Parsons, Cameron and Lee, Sangmi and Jayeola, Victor and Kathariou, Sophia}, year={2017}, month={Mar} } @article{martinez_osborne_jayeola_katic_kathariou_2016, title={Capacity of Listeria monocytogenes Strains from the 2011 Cantaloupe Outbreak To Adhere, Survive, and Grow on Cantaloupe}, volume={79}, ISSN={["1944-9097"]}, DOI={10.4315/0362-028x.jfp-15-498}, abstractNote={The 2011 listeriosis outbreak attributed to whole cantaloupe involved several genetically distinct strains of serotypes 1/2a and 1/2b that had not been previously reported in invasive listeriosis outbreaks. Here we investigated the potential of strains from the 2011 cantaloupe outbreak to adhere, survive, and grow on cantaloupe rind and flesh and in juice extracted from cantaloupe at different temperatures (4, 8, and 25°C). All strains were able to adhere and grow, with ∼10-fold increases after 7 days at 4 or 8°C and after 24 h at 25°C, with a propensity for more growth on rind than on flesh or in extract. No significant differences in growth potential were noted among the different strains or between them and unrelated strains from other listeriosis outbreaks involving celery, deli meats, or hot dogs. Similarly to the cantaloupe outbreak strains, these other strains exhibited greater propensity for growth on rind than on flesh or in extract. Rinsing of cantaloupe fragments in sterile water resulted in temporary reductions of the populations by 50- to 100-fold, suggesting the potential of such washing to reduce risk if the produce is promptly consumed. The absence of marked differences in adherence or growth between the cantaloupe outbreak strains and strains from other outbreaks highlights the need to further characterize the 2011 cantaloupe outbreak strains and elucidate potential biological attributes that contributed to their implication in the outbreak.}, number={5}, journal={JOURNAL OF FOOD PROTECTION}, author={Martinez, Mira Rakic and Osborne, Jason and Jayeola, Victor Oladimeji and Katic, Vera and Kathariou, Sophia}, year={2016}, month={May}, pages={757–763} }