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Cronobacter is an important foodborne pathogen that can cause severe neonatal meningitis, necrotizing enterocolitis, and bacteremia. Currently, there is limited knowledge of biofilm formation in Cronobacter. In the present study, biofilm formation ability and associated gene expression changes in Cronobacter from cereal related samples was carried out systematically. Our results from 307 Cronobacter isolates analyzed for 48 h showed strong biofilm-forming ability in 14 strains (4.6%), moderate in 47 strains (15.3%), weak in 142 strains (46.2%), and no such ability in the remaining 104 strains (33.9%). Further studies on five strains with strong biofilm-forming ability showed that maximum biofilm formation in Cronobacter occurred after 24 h of cultivation, reaching a peak around 48 h–72 h, reducing gradually thereafter. Kyoto encyclopedia of genes and genomes (KEGG) analysis revealed that differentially expressed genes (DEGs) involved in flagellar assembly, oxidative phosphorylation, ribosome, photosynthesis, O-Antigen nucleotide sugar biosynthesis, citrate cycle (tricarboxylic acid cycle, TCA) and bacterial chemotaxis were enriched in biofilm forming cells. The genes involved these enrichment pathways were mostly downregulated when compared to planktonic cells. Several transcriptional regulator genes such as csrA and bolA , and the cell surface composition regulator gene glgS were significantly upregulated. 12 of 13 (92.3%) selected genes was found to be in agreement with the RNA-Seq of planktonic and biofilm cells by Quantitative real-time PCR analysis, thus increasing confidence in our data. Our research lays a sound theoretical basis for further studies on mechanisms regulating biofilm formation and provides a foundation for development of new food safety measures, clinical disease prevention and control. [Display omitted] • Evaluate biofilm formation ability in 307 Cronobacter from cereal related samples • Elucidate the dynamic process of biofilm formation in selected five Cronobacter • Clarify the transcriptome difference of planktonic and biofilm cells of Cronobacter • Predict the genes and pathways that might affect biofilm formation of Cronobacter [ABSTRACT FROM AUTHOR]

Abstract Cronobacter are opportunistic bacterial pathogens of both infants and adults. We investigated the incidence and distribution of Cronobacter in 1245 samples of cereal and related environments. 39.1% (101/258) rice-related and 46.9% (98/209) wheat-related samples tested positive for Cronobacter , and the positive rate differed notably according to processing method. Cronobacter was found in rice and wheat plants at the tillering, filling and mature stages. Soil, water and swab samples from nearby milling plants were assayed, and results revealed that 6.3% (7/122) of water from paddy fields, 49.1% (28/57) and 62.1% (41/67) of swab samples from rice and wheat flour milling plants were Cronobacter positive. Pulsed-field gel electrophoresis (PFGE) subtyping indicated that some strains had a common profile, which suggested their persistence in the environment, potential transmission routes and cross-contamination in processing. Finally, we surveyed 18 families to evaluate potential risks. None of the families who primarily ate rice cooked with water tested positive for Cronobacter , though of 66.7% families (6/9) whose food staples were produced from wheat flour tested positive. Taken together, our results are important for understanding Cronobacter transmission and will aid in the development of additional control measures to reduce the risk of infection by these opportunistic pathogenic bacteria. Highlights • Cronobacter detection rates notably changed with different processing methods. • Cronobacter was present in different growth stages of rice and wheat. • The presence of Cronobacter in the domestic environment was related to primary staple foods. • Consumption of cereal flour may pose a high risk for Cronobacter infection. [ABSTRACT FROM AUTHOR]

Cronobacter was positive in cereals at a relatively high rate. In the present study, we investigated the occurrence and characteristics of this pathogen systematically in diverse cereals. All sampled food (N = 467) contained Cronobacter with a high positive rate of 54.0%. The enumeration experiment showed the concentration ranged from 0.3 to more than 110 MPN/100 g, and 87.9% of 127 samples were less than 10 MPN/100 g. There were significant differences (p < 0.05) in positive rates for Cronobacter among cereal kernels (40.2%), cereal flour (66.7%), cereal products made from raw cereal flour (87.6%), and cereal products made from flour (ready-to-eat) (17.4%). The dominant Cronobacter species was C. sakazakii and C. dublinensis , followed by C. malonaticus and C. turicensis. Two interesting clusters with more than 90% similarities were identified by pulsed-field gel electrophoresis (PFGE). The C1 cluster (four isolates) indicated these strains were derived from a common source and persisted in the food production environment for an extended time. The C2 cluster (six isolates) indicated the pathogen could be transmitted via cereal processing. Our research provided baseline data for Cronobacter in diverse cereals and was helpful for understanding Cronobacter transmission. The results also indicate that additional control measures should be developed to reduce the risk of infection by these opportunistic pathogenic bacteria. • A high positive rate (54.0%) of Cronobacter was found in diverse cereals. • Cereal flour might pose a risk of Cronobacter infection. • The dominant species are C. sakazakii , C. dublinensis , and C. Malonaticus. • The pathogen could be transmitted via cereal processing. [ABSTRACT FROM AUTHOR]

The common use of gloves in retail practices represents a potential route for cross contamination of foodborne pathogens in fresh-cut produce. Using fresh-cut cantaloupe as a food model, we investigated factors that may influence glove-mediated cross contamination by Listeria monocytogenes and developed mathematical models to illustrate the patterns of transfer during fresh-cutting practices. Contact time (2, 5, 10 s), contact pressure (0.05, 0.18, 0.37 psi), and glove type (nitrile, polyvinyl chloride, polyethylene) did not have a significant effect on transfer of L. monocytogenes from cantaloupe rind to flesh, or from flesh to flesh. However, glove type appeared to affect L. monocytogenes transfer from the stem scar tissue to cantaloupe flesh (P = 0.0371). Transfer from rind pieces that had been washed with water was significantly higher than transfer from pieces that had not been washed (P = 0.0006). Predictive modeling and experimental validation suggested that transfer of L. monocytogenes on cantaloupe flesh persists over 85 pieces through consecutive contacts with a gloved hand. Findings of the study provide new scientific data to aid researchers, retailers, and caterers in safety risk assessments of fresh-cut practices used to prepare cantaloupes and other produce items. • Single-use gloves for preparation of fresh-cut cantaloupes were possible vehicles for sustained transfer of L. monocytogenes. • Multiple factors were investigated for their impact on glove-mediated cross contamination of fresh-cut cantaloupe by L. monocytogenes. • Residue surface moisture significantly facilitated the transfer of L. monocytogenes from cantaloupe rind to flesh via gloves. • Mathematical models were developed to illustrate L. monocytogenes transfer on fresh-cut cantaloupe through gloved hands. [ABSTRACT FROM AUTHOR]