One Health approach for prioritization of potential foodborne pathogens: Risk-ranking, Delphi survey, and criteria evaluation pre- and post-COVID-19 pandemic.

Frequent foodborne illnesses with unknown causative agents highlight the need to explore zoonotic potential foodborne pathogens (PFPs). An effective PFP prioritization tool is indispensable, especially after experiencing the recent pandemic caused by zoonotic SARS-CoV-2. Risk information on pathogens (excluding 30 known foodborne pathogens) provided by governmental and international organizations was reviewed to generate a list of PFPs. Risk-ranking of PFPs was conducted based on a literature review of food poisoning or detection cases, and the ranks were determined with a decision tree. PFPs were prioritized by infectious disease (ID), veterinary medicine (VET), and food safety (FS) experts through a pre- and postpandemic Delphi survey, and key criteria in their decisions were illuminated. Among 339 PFPs, 32 rank-1 PFPs were involved in the foodborne outbreak(s). Discrepancies in opinions on prioritization between experts in different fields deepened after the pandemic. Only VET and FS experts valued the plausibility of foodborne transmission in evaluating bacteria and viruses, and a significant correlation between their selection of PFPs was found (p < .05). The impact of the pandemic induced all fields to focus more on human transmission and severity/fatality in prioritizing viruses, and only FS experts emphasized the plausibility of foodborne transmission after the pandemic. In contrast to prioritizing bacteria or viruses, ID and VET experts are unusually focused on foodborne transmission when prioritizing parasites. Criteria of consensus deduced by interdisciplinary experts with different interests and the criteria directly related to foodborne transmission should be acknowledged for adequate PFP prioritization.

Microbial succession during button mushroom (Agaricus bisporus) production evaluated via high-throughput sequencing.

Button mushrooms (Agaricus bisporus), are one of the most widely consumed mushrooms in the world. However, changes within its microbial community as it relates to the use of different raw materials and cultivation methods, as well as potential points of microbial contamination throughout the production process have not been investigated extensively. In the present study, button mushroom cultivation was investigated in each of the four stages (raw materials, composting (phase I, Ⅱ, and Ⅲ), casing, and harvesting), and samples (n = 186) from mushrooms and their related environments were collected from four distinct mushroom-growing farms (A-D) in Korea. Shifts within the bacterial consortium during mushroom production were characterized with 16 S rRNA amplicon sequencing. The succession of bacterial communities on each farm was dependent on the raw material incorporated, aeration, and the farm environment. The dominant phyla of the compost stack at the four farms were Pseudomonadota (56.7%) in farm A, Pseudomonadota (43.3%) in farm B, Bacteroidota (46.0%) in farm C, and Bacillota (62.8%) in farm D. During the Phase Ⅰ, highly heat-resistant microbes, such as those from the phylum Deinococcota (0.6-65.5%) and the families Bacillaceae (1.7-36.3%), Thermaceae (0.1-65.5%), and Limnochordaceae (0.3-30.5%) greatly proliferated. The microbial diversity within compost samples exhibited a marked decline as a result of the proliferation of thermophilic bacteria. In the spawning step, there were considerable increases in Xanthomonadaceae in the pasteurized composts of farms C and D - both of which employed an aeration system. In the harvesting phase, beta diversity correlated strongly between the casing soil layer and pre-harvest mushrooms, as well as between gloves and packaged mushrooms. The results suggest that gloves may be a major source of cross-contamination for packaged mushrooms, highlighting the need for enhanced hygienic practices during the harvesting phase to ensure product safety. These findings contribute to the current understanding of the influence of environmental and adjacent microbiomes on mushroom products to benefit the mushroom industry and relevant stakeholders by ensuring quality production.

Genotypes of Blastocystis sp. among elderly health checkup people in South Korea with a questionnaire on risk factors.

Blastocystis sp. is a common zoonotic intestinal parasite of humans and animals, and has been classified into at least 17 distinct subtypes. Despite its potential impact on public health, the prevalence and subtype distribution of Blastocystis sp. have seldom been the study subject in South Korea. To determine the prevalence and subtype distribution of Blastocystis sp. and to obtain information on risk factors, we performed a cross-sectional study targeting elderly health checkup people, who visited Seoul Western Branch of the Korea Association of Health Promotion (KAHP) in October 2019. Stool samples were collected from 293 participants consisting of 128 males and 165 females with a mean age of 64.7 years (from 50 to 88 years) with a questionnaire on potential risk factors. All the samples were tested by PCR targeting the SSU rRNA gene of Blastocystis sp., and nucleotide sequences of positive samples were used to identify the subtypes. The overall prevalence of Blastocystis sp. was 9.2% (27/293). Among the positive samples, subtype 3 was predominant (59%; 16/27), followed by subtype 1 (41%; 11/27). No other subtypes were detected. In the univariable analysis, the age, sex, presence of digestive symptoms, source of drinking water, and history of drug intake were not significantly associated with Blastocystis sp. infection. Two parameters, including the Enterococcus hirae bacterial infection and the frequency of intake of cooked or boiled vegetables less than twice a week, showed statistical significance. However, the multivariable regression analysis revealed that only the latter parameter was statistically significant. The results suggested that subtypes 3 and 1 are the 2 major genotypes of Blastocystis sp. among elderly people in South Korea, and low frequency of consuming cooked or boiled vegetables is a potential risk factor.

Recent Advances in the Application of Antibacterial Complexes Using Essential Oils.

Although antibacterial spectrum of essential oils (EOs) has been analyzed along with consumers' needs on natural biocides, singular treatments generally require high concentration of EOs and long-term exposures to eliminate target bacteria. To overcome these limitations, antibacterial complex has been developed and this review analyzed previous reports regarding the combined antibacterial effects of EOs. Since unexpectable combined effects (synergism or antagonism) can be derived from the treatment of antibacterial complex, synergistic and antagonistic combinations have been identified to improve the treatment efficiency and to avoid the overestimation of bactericidal efficacy, respectively. Although antibacterial mechanism of EOs is not yet clearly revealed, mode of action regarding synergistic effects especially for the elimination of pathogens by using low quantity of EOs with short-term exposure was reported. Whereas comprehensive analysis on previous literatures for EO-based disinfectant products implies that the composition of constituents in antibacterial complexes is variable and thus analyzing the impact of constituting substances (e.g., surfactant, emulsifier) on antibacterial effects is further needed. This review provides practical information regarding advances in the EO-based combined treatment technologies and highlights the importance of following researches on the interaction of constituents in antibacterial complex to clarify the mechanisms of antibacterial synergism and/or antagonism.

Short-Term Antifungal Treatments of Caprylic Acid with Carvacrol or Thymol Induce Synergistic 6-Log Reduction of Pathogenic Candida albicans by Cell Membrane Disruption and Efflux Pump Inhibition.

BACKGROUND/AIMS: Although naturally-derived antifungals have been investigated for their ability to inactivate Candida albicans, which is a major cause of candidiasis, they have shown a less than 3 log reduction in C. albicans or required treatment times of longer than 3 h. Thus, the naturally-derived antifungals used in previous studies could not substantially eradicate C. albicans within a short period of time. METHODS: To improve the fungicidal effects of naturallyderived antifungals against C. albicans within short time periods, we developed composites showing antifungal synergism using caprylic acid (CA), carvacrol (CAR) and thymol (THM) for 1-10 min at 22/37°C. Using flow cytometry, we examined the mode of action for the synergism of these compounds on membrane integrity and efflux pump activity. RESULTS: Whereas the maximum reduction by individual treatments was 0.6 log CFU/ml, CA + CAR/THM (all 1.5 mM) eliminated all pathogens (> 6.8 log reduction) after 1 min at 37°C and after 10 min at 22°C. The flow cytometry results showed that exposure to CA damaged the membranes in 15.7-36.5% of cells and inhibited efflux pumps in 15.4-31.3% of cells. Treatments with CAR/THM slightly affected cell membranes (in 1.8-6.9% of cells) but damaged efflux pumps in 14.4-29.6% of cells. However, the combined treatments clearly disrupted membranes (> 83.1% of cells) and pumps (> 95.0% of cells). The mechanism of this synergism may involve membrane damage by CA, which facilitates the entry of antifungals into the cytoplasm, and the inhibition of efflux pumps by CA, CAR or THM, causing their accumulation within cells and, leading to cell death. CONCLUSION: Antifungal composites (CA + CAR/THM) showing synergism (i.e., an additional 6 log reduction) within minutes at room/body temperature can be used to treat candidiasis and improve the microbiological safety of facilities contaminated with fungi as a novel alternative to synthetic antifungals.

Health Functionality and Quality Control of Laver (Porphyra, Pyropia): Current Issues and Future Perspectives as an Edible Seaweed.

The growing interest in laver as a food product and as a source of substances beneficial to health has led to global consumer demand for laver produced in a limited area of northeastern Asia. Here we review research into the benefits of laver consumption and discuss future perspectives on the improvement of laver product quality. Variation in nutritional/functional values among product types (raw and processed (dried, roasted, or seasoned) laver) makes product-specific nutritional analysis a prerequisite for accurate prediction of health benefits. The effects of drying, roasting, and seasoning on the contents of both beneficial and harmful substances highlight the importance of managing laver processing conditions. Most research into health benefits has focused on substances present at high concentrations in laver (porphyran, Vitamin B12, taurine), with assessment of the expected effects of laver consumption. Mitigation of chemical/microbiological risks and the adoption of novel technologies to exploit under-reported biochemical characteristics of lavers are suggested as key strategies for the further improvement of laver product quality. Comprehensive analysis of the literature regarding laver as a food product and as a source of biomedical compounds highlights the possibilities and challenges for application of laver products.

Underrecognized niche of spore-forming bacilli as a nitrite-producer isolated from the processing lines and end-products of powdered infant formula.

Although nitrite in powdered milk formula (PIF) is a recognized health risk for infants, the presence of nitrite in PIF has only been investigated as a chemical contaminant during the inspection of end-products. The risk posed by microbial sources of nitrite during the PIF manufacturing process has not been considered. This is the first study to report the taxonomy and physiological characteristics of nitrite-producing bacteria isolated from PIF processing environments. All isolates identified as nitrite-producers (133 out of 501 strains collected over four years) from work-in-process and end-products of PIF were spore-forming bacilli. Nitrite-producing metabolism under PIF processing conditions was found in not only thermophilic isolates (3 Bacillus, 60 Geobacillus from 63 strains; 100%) but also in mesophilic isolates (65 Bacillus, 1 Anoxybacillus from 70 strains; 65.7%). Geobacillus was the only highly heat-resistant sporeformer and vigorous nitrite-producer exhibiting dramatic increases in nitrite over short periods of incubation (a maximum value within 3 h). High conversions of nitrate to nitrite (up to 88.8%) was also observed, highlighting bacteria as a key source of nitrite in PIF processing lines. Further research into the diversity of metabolic activity observed in this study can facilitate specialized management of nitrite-producers in PIF processing lines.

Prevalence of pathogenic Arcobacter species in South Korea: Comparison of two protocols for isolating the bacteria from foods and examination of nine putative virulence genes.

Contamination of foodstuffs by potentially enteropathogenic Arcobacter spp. is becoming a concern worldwide. However, few studies have examined virulence-associated genes in isolates of Arcobacter spp. from food. Here, we investigated the prevalence of three pathogenic Arcobacter species, A. butzleri, A. cryaerophilus, and A. skirrowii, in chicken, pork, and leafy green vegetables (n = 323) in South Korea. Samples were examined using two different protocols selected from a literature review: Acrobacter selective broth (ASB) II + Arcobacter selective medium (ASM) II (protocol A), and ASB II + modified charcoal cefoperazone deoxycholate agar supplemented with CAT (protocol B). Overall, Arcobacter spp. were detected in 45.8% of food samples, and the recovery rate of protocol B (37.8%) was significantly higher than that of protocol A (30.7%) (p < 0.05). Refrigerated chicken gizzard samples showed the highest detection rate (100%), followed by refrigerated chicken wing (79.5%), intestine (77.3%), neck skin (63.3%), pork (55.6%), frozen chicken legs (5.0%), and leafy green vegetables (4.4%) (p < 0.05). All isolates from chicken and leafy green vegetables were identified as A. butzleri, whereas A. cryaerophilus and A. skirrowii were mainly detected in pork. Most samples (95.8%) harbored more than one of nine putative virulence factors (cadF, ciaB, cj1349, hecA, hecB, mviN, pldA, irgA, and tlyA), and 91.3% harbored more than two. Isolates harboring all nine putative virulence genes were obtained from 1.9% of samples: five pork and one chicken. This study provides comprehensive and de facto evidence regarding prevalence of an emerging pathogen, Arcobacter spp., in various foods, along with their virulence potential. The results justify further research with respect to their role in food safety.

Response surface modeling of reductions in uropathogenic Escherichia coli biofilms on silicone by cranberry extract, caprylic acid, and thymol.

A response surface methodology was used to build a model to predict reductions in uropathogenic Escherichia coli biofilms in response to three compounds: cranberry extract [CB] at 3.0-9.0%, and caprylic acid [CAR] and thymol [TM] at 0.01%-0.05%. The predictive model for microbial reduction had a high regression coefficient (R2 = 0.9988), and the accuracy of the model was verified (R2 = 0.9527). Values of CAR, TM, and the quadratic term CAR2 were the most significant (P < 0.0001) for bacterial reduction. Interactions between CB and CAR, and TM and CB, also affected bacterial reduction. The optimum conditions (a 5.8 log10 reduction) determined by ridge analysis were 8.3% CB +0.04% CAR +0.04% TM at 37 °C for 1 min. The model could be used to predict the most cost-efficient amounts of antimicrobial agents for anti-urinary tract infection products such as catheter lock solution and antimicrobial coatings for catheters.

Microbial diversities and potential hazards of Korean turbid rice wines (makgeolli): Multivariate analyses.

A number of makgeolli (Korean traditional turbid rice wine) products are commercially available in various forms. To date, there has been no comprehensive investigation of these products. Here, we collected samples of almost all of the makgeolli products that are currently commercially available (n = 167), recorded their manufacturing variables, and examined physiochemical parameters and microbial communities, using quantitative and qualitative methods. The aerobic plate count (APC) and counts of lactic acid bacteria (LAB), acetic acid bacteria (AAB), fungi, total coliforms, and Bacillus cereus were obtained, and the presence of Escherichia coli and eight foodborne pathogens was also examined. The data obtained were segmented and analyzed based on multiple variables associated with the manufacturing characteristics. Despite high ethanol contents (up to 16.0%) and high acidities (pH 3.3-4.5), the rice wine products examined here had diverse and abundant microbiotas (mean values: APC, 5.3; LAB,4.4; AAB,1.5; fungi, 3.8 log CFU/ml). In particular, LAB and fungi, which are used as co-starter cultures during rice wine manufacturing, accounted for the majority of the microbiotas. Bivariate analyses revealed significant positive correlations between the individual micro-organism counts (Pearson correlation coefficient = 0.668-0.947). Among the manufacturing variables considered in this study, only the pasteurization status had a significant effect on the microbial communities of rice wine products (p < 0.05). When examining the presence of foodborne pathogens, B. cereus was isolated from some of the rice wine products (58.1%) at low levels (<100 CFU/ml), and its detection rate was not significantly lower in the pasteurized products than the raw products. Overall, the results presented here provide a comprehensive overview of the microbiotas of commercially available turbid rice wines and their relationships to manufacturing variables. These data will help to direct future studies focusing on rice wine quality and safety control measures.

Current Interventions for Controlling Pathogenic Escherichia coli.

This review examined scientific reports and articles published from 2007 to 2016 regarding the major environmental sources of pathogenic Escherichia coli and the routes by which they enter the human gastrointestinal tract. The literature describes novel techniques used to combat pathogenic E. coli transmitted to humans from livestock and agricultural products, food-contact surfaces in processing environments, and food products themselves. Although prevention before contamination is always the best "intervention," many studies aim to identify novel chemical, physical, and biological techniques that inactivate or eliminate pathogenic E. coli cells from breeding livestock, growing crops, and manufactured food products. Such intervention strategies target each stage of the food chain from the perspective of "Farm to Table food safety" and aim to manage major reservoirs of pathogenic E. coli throughout the entire process. Issues related to, and recent trends in, food production must address not only the safety of the food itself but also the safety of those who consume it. Thus, research aims to discover new "natural" antimicrobial agents and to develop "multiple hurdle technology" or other novel technologies that preserve food quality. In addition, this review examines the practical application of recent technologies from the perspective of product quality and safety. It provides comprehensive insight into intervention measures used to ensure food safety, specifically those aimed at pathogenic E. coli.

Sodium Chloride Does Not Ensure Microbiological Safety of Foods: Cases and Solutions.

Addition of salt or salt-containing water to food is one of the oldest and most effective preservation methods in history; indeed, salt-cured foods are generally recognized as microbiologically safe due to their high salinity. However, a number of microbiological risks remain. The microbiological hazards and risks associated with salt-cured foods must be addressed more in-depth as they are likely to be underestimated by previous studies. This review examined a number of scientific reports and articles about the microbiological safety of salt-cured foods, which included salted, brined, pickled, and/or marinated vegetables, meat, and seafood. The following subjects are covered in order: (1) clinical cases and outbreaks attributed to salt-cured foods; (2) the prevalence of foodborne pathogens in such foods; (3) the molecular, physiological, and virulent responses of the pathogens to the presence of NaCl in both laboratory media and food matrices; (4) the survival and fate of microorganisms in salt-cured foods (in the presence/absence of additional processes); and (5) the interaction between NaCl and other stressors in food processes (e.g., acidification, antimicrobials, drying, and heating). The review provides a comprehensive overview of potentially hazardous pathogens associated with salt-cured foods and suggests further research into effective intervention techniques that will reduce their levels in the food chain.

Changes in microbial composition and the prevalence of foodborne pathogens in crab marinated in soy sauce produced by six manufacturing plants.

BACKGROUND: The present study examined the changes in microbiological composition during the production process of crab marinated in soy sauce, potential microbial hazards, potential contamination routes and effective critical control points. Crab and soy sauce samples were obtained from six different manufacturing plants at different stages, and their microbiological content was comprehensively assessed by quantitative and qualitative analyses. RESULTS: The results revealed the following: (1) the final products contained 4.0 log colony-forming units (CFU) g-1 aerobic plate counts (APCs) and 1.1 log CFU g-1 coliforms, which may have been introduced from the raw materials (the level of APCs in raw crab and soy sauce mixed with other ingredients was 3.8 log CFU g-1 and 4.0 log CFU mL-1 respectively); (2) marination of crab in soy sauce may allow cross-contamination by coliforms; (3) only Bacillus cereus and Staphylococcus aureus were qualitatively detected in samples at different stages of manufacture (detection rate of 28 and 5.6% respectively), and these bacteria may impact the microbiological quality and safety of crab marinated in soy sauce; and (4) bacterial counts were either maintained or increased during the manufacturing process (suggesting that no particular step can be targeted to reduce bacterial counts). CONCLUSION: Proper management of raw materials and the marination process are effective critical control points, and alternative interventions may be needed to control bacterial quantity. The results provide important basic information about the production of crab marinated in soy sauce and may facilitate effective implementation of sanitary management practices in related industries and research fields. © 2016 Society of Chemical Industry.

Phytic Acid and Sodium Chloride Show Marked Synergistic Bactericidal Effects against Nonadapted and Acid-Adapted Escherichia coli O157:H7 Strains.

The synergistic antimicrobial effects of phytic acid (PA), a natural extract from rice bran, plus sodium chloride against Escherichia coli O157:H7 were examined. Exposure to NaCl alone at concentrations up to 36% (wt/wt) for 5 min did not reduce bacterial populations. The bactericidal effects of PA alone were much greater than those of other organic acids (acetic, citric, lactic, and malic acids) under the same experimental conditions (P < 0.05). Combining PA and NaCl under conditions that yielded negligible effects when each was used alone led to marked synergistic effects. For example, whereas 0.4% PA or 3 or 4% NaCl alone had little or no effect on cell viability, combining the two completely inactivated both nonadapted and acid-adapted cells, reducing their numbers to unrecoverable levels (>7-log CFU/ml reduction). Flow cytometry confirmed that PA disrupted the cell membrane to a greater extent than did other organic acids, although the cells remained viable. The combination of PA and NaCl induced complete disintegration of the cell membrane. By comparison, none of the other organic acids acted synergistically with NaCl, and neither did NaCl-HCl solutions at the same pH values as the test solutions of PA plus NaCl. These results suggest that PA has great potential as an effective bacterial membrane-permeabilizing agent, and we show that the combination is a promising alternative to conventional chemical disinfectants. These findings provide new insight into the utility of natural compounds as novel antimicrobial agents and increase our understanding of the mechanisms underlying the antibacterial activity of PA.

Estimation of Sensory Pork Loin Tenderness Using Warner-Bratzler Shear Force and Texture Profile Analysis Measurements.

This study investigated the degree to which instrumental measurements explain the variation in pork loin tenderness as assessed by the sensory evaluation of trained panelists. Warner-Bratzler shear force (WBS) had a significant relationship with the sensory tenderness variables, such as softness, initial tenderness, chewiness, and rate of breakdown. In a regression analysis, WBS could account variations in these sensory variables, though only to a limited proportion of variation. On the other hand, three parameters from texture profile analysis (TPA)-hardness, gumminess, and chewiness-were significantly correlated with all sensory evaluation variables. In particular, from the result of stepwise regression analysis, TPA hardness alone explained over 15% of variation in all sensory evaluation variables, with the exception of perceptible residue. Based on these results, TPA analysis was found to be better than WBS measurement, with the TPA parameter hardness likely to prove particularly useful, in terms of predicting pork loin tenderness as rated by trained panelists. However, sensory evaluation should be conducted to investigate practical pork tenderness perceived by consumer, because both instrumental measurements could explain only a small portion (less than 20%) of the variability in sensory evaluation.

Enhanced protection of pathogenic Escherichia coli ingested by a soil nematode Caenorhabditis elegans against sanitizer treatments.

We employed Caenorhabditis elegans as a model to study the effectiveness of sanitizers in killing pathogenic Escherichia coli strains ingested by free-living nematodes. Adult worms that had fed on six pathogenic E. coli strains (highly persistent in the nematode intestine) were treated with three chemical solutions. In planktonic cells, none of the H2O2 and acetic acid treatments influenced the survival of the pathogenic E. coli strains, whereas sodium hypochlorite critically decreased the viability of the strains. Importantly, the survival of the E. coli strains was dramatically increased by persistence in the C. elegans gut under 0.1% sodium hypochlorite, and several strains could survive at a concentration of 0.5%. In addition, all pathogenic E. coli strains in the C. elegans gut survived on the lettuce for 5 days even though they were washed with 0.1% sodium hypochlorite. Taken together, our results indicate that pathogenic E. coli ingested by C. elegans may be protected against washing treatment with commercial sanitizers on raw food materials.

Novel hypothesis for infant methemoglobinemia: Survival and metabolism of nitrite-producers from vegetables under gastrointestinal stress and intestinal adhesion.

Infants have digestive environments that are more favorable for microbial proliferation and subsequent endogenous nitrite production than those of adults, but direct evidence of this has been lacking. In this study, we propose a novel epidemiology of infant methemoglobinemia by demonstrating the risk posed by nitrite-producers in the gastrointestinal tract. Nitrite-producers from vegetables (n = 323) were exposed to stress factors of the gastrointestinal environment (gastric pH, intestinal bile salts, anaerobic atmosphere) reflecting 4 different postnatal age periods (Neonate, ≤1 month; Infant A, 1-3 months; Infant B, 3-6 months; Infant C, 6-12 months). "High-risk" strains with a nitrate-to-nitrite conversion rate of ≥1.3 %, the minimum rate corresponding to nitrite overproduction, under the Neonate stress condition were analyzed for intestinal adhesion. Among all the phyla, Pseudomonadota achieved the highest survival (P < 0.05; survival rate of 51.3-71.8 %). Possible cross-protection against bile resistance due to acid shock was observed for all the phyla. All the high-risk strains exhibited moderate autoaggregation (14.0-36.4 %), whereas only a few exhibited satisfactory surface hydrophobicity (>40 %). The Pantoea agglomerans strain strongly adhered to Caco-2 cells (7.4 ± 1.1 %). This study showed the ability of the Pantoea, Enterobacter, and Klebsiella strains to survive under gastrointestinal stress for ≤12 months, to excessively produce nitrite under neonatal stress conditions, and to settle in the human intestine. To our knowledge, this is the first study to reveal the role of the natural flora of vegetables in the epidemiology of infant methemoglobinemia through a multilateral approach.

Prevalence and phylogenetic traits of nitrite-producing bacteria in raw ingredients and processed baby foods: Potential sources of foodborne infant methemoglobinemia.

Nitrite, which has been mainly regarded as a chemical hazard, can induce infant methemoglobinemia. As for nitrite as a product of microbial metabolism, the contribution of the oral or gut microbiome has mostly received attention, whereas the role of nitrite-producing bacteria (NPBs) in food has been less elucidated. In this study, mesophilic NPBs were isolated from food samples (n = 320) composed of raw ingredients for weaning foods (n = 160; beetroot, broccoli, carrot, lettuce, rice powder, spinach, sweet potato, and honey) and processed baby foods (n = 160; cereal snack, cheese, yogurt, powdered infant formula, sorghum syrup, vegetable fruit juice, and weaning food). The phylogenetic diversity of the NPB strains was analyzed via 16S rRNA sequencing. All 15 food items harbored NPBs, with a prevalence of 71.9 % and 34.4 % for the raw ingredients and processed foods, respectively. The NPBs isolated from the foods were identified as Actinomycetota (Actinomycetes), Bacteroidota (Flavobacteriia, Sphingobacteriia), Bacillota (Bacilli), or Pseudomonadota (Alpha-, Beta-, and Gammaproteobacteria). Among the raw and processed foods, beetroot (85.0 %) and powdered infant formula (70.0 %) showed had the highest NPB prevalence (P > 0.05). Bacillota predominated in both types of food. The contamination source of Pseudomonadota, which was another major phylum present in the raw ingredients, was presumed to be the soil and endophytes in the seeds, whereas that of Bacillota was the manufacturing equipment used with the raw ingredients. Common species for probiotics, such as Lacticaseibacillus, Leuconostoc, Enterococcus, and Bacillus, were isolated and identified as NPBs. To our knowledge, this is the first study to reveal the taxonomical diversity and omnipresence of NPBs in food for babies. The results of this study highlight the importance of food-mediated microbiological risks of infant methemoglobinemia which are yet underrecognized.

A rapid and simple screening method of Cronobacter spp. in cell suspension and tofu.

BACKGROUND: Cronobacter spp. causes illnesses such as neonatal meningitis, sepsis and necrotising enterocolitis that can be fatal in infants and neonates. Thus it is of great concern to the food industry, which requires a simple and miniaturised method for the rapid detection of Cronobacter spp. in food. RESULTS: A simple and rapid method was developed for the quantitative estimation of Cronobacter spp. Cell suspension cultures and tofu inoculated with Cronobacter spp. as well as a mixed culture (Cronobacter spp. and other micro-organisms) were serially diluted twofold on microtitre plates. Kim and Rhee broth, containing salicin, was added to each well. After 16-18 h of incubation the colour of broth in the wells changed from purple to yellow owing to salicin fermentation by Cronobacter spp. Bacterial counts were predicted by analysing the end points (the last well number in a series of positive wells). The coefficients of determination (r(2)) between the conventional plating method and the current method had high degrees of correlation (0.954-0.963), suggesting that the latter method offers a reasonable quantification of Cronobacter spp. CONCLUSION: The simple method developed in this study could be used for the efficient and rapid screening of Cronobacter spp. in the food industry.

Characterization of ready-to-eat fish surface as a potential source of contamination of Vibrio parahaemolyticus biofilms.

The worldwide consumption of ready-to-eat seafood products has steadily increased due to a range of health benefits. However, depending on the handling or cutting process of raw fish, ready-to-eat sashimi can be exposed to microbiological risks that can lead to foodborne infection by marine pathogens. Since surface characteristics are key factors for microbial adhesion and biofilm formation, the present study aims to determine the correlation between raw fish skin properties and Vibrio parahaemolyticus biofilm formation. We analyzed V. parahaemolyticus biofilms (ATCC 17802; initially inoculated ca. 2 or 4 log CFU/cm2) on fish skin (gizzard shad, pomfret, red snapper, and mackerel; fish species served as sashimi without peeling the skin) formed under simulated marine environments (incubating in artificial seawater with rocking motion at 30 °C, the maximum temperature of seasonal seawater) for 24 h. The characteristics of fish skin were determined using confocal laser scanning microscopy/scanning electron microscopy. V. parahaemolyticus showed higher biofilm counts on fish skins than on stainless steel, which was used as a control (P < 0.05). In particular, V. parahaemolyticus formed biofilms with significantly higher levels of bacterial populations on gizzard shad and pomfret (ca. 5 log CFU/cm2; P < 0.05), highlighting the relationship between the biofilm formation level and the characteristics of gizzard shad and pomfret skins. The surface roughness of fish skins, including the main roughness parameters (Ra, Rq, and Rz), influenced the attachment of V. parahaemolyticus (P < 0.05). Additionally, images of V. parahaemolyticus biofilms suggested that different topographical profiles of fish species (e.g., mucus, unique structural features, etc.) could cause V. parahaemolyticus to exhibit different biofilm phenotypes, such as sticking to or entangling on the fish skin surface. The major findings of this study provide various phenotypic adhesions of V. parahaemolyticus to fish skin in considerations of the potential hazard for the consumption of ready-to-eat sashimi served with its skin.