Sodium Hypochlorite (NaClO) Disturbed Lipid Metabolism in Larval Zebrafish (Danio rerio), as Revealed by Lipidomics and Transcriptomics Analyses.

Sodium hypochlorite (NaClO) has been widely utilized since the initial outbreak of coronavirus disease (COVID-19). The widespread use of NaClO means that it can directly enter aquatic ecosystems through wastewater discharge. In this study, we analyzed the expression of PPAR-γ, FAS, and ACC1, which significantly increased in larval zebrafish following exposure to 300 µg/L NaClO for 7 days. Additionally, we examined the effects of high concentrations of NaClO on zebrafish through non-targeted lipidomics and transcriptomics. A total of 44 characteristic lipid molecules were identified using non-targeted lipidomics; an absolute quantitative analysis revealed that the contents of these subclasses of lipids decreased significantly following exposure to 300 µg/L NaClO for 7 days. The levels of triglyceride (TG), phosphatidylethanolamines (PE), and diglyceride (DG) were particularly affected. Transcriptomic analysis revealed that exposure to 300 µg/L NaClO could significantly disrupt global gene transcription in larval zebrafish. Interestingly, more than 700 differentially expressed genes (DEGs) were identified, primarily associated with lipid metabolism and glycometabolism pathways. Overall, our study provided new insights into the toxicological effects of chlorine-containing disinfectants in aquatic organisms.

Unraveling the spoilage characteristics of refrigerated pork using high-throughput sequencing coupled with UHPLC-MS/MS-based non-targeted metabolomics.

The spoilage of refrigerated pork involves nutrient depletion and the production of spoilage metabolites by spoilage bacteria, yet the microbe-metabolite interactions during this process remain unclear. This study employed 16S rRNA high-throughput sequencing and non-targeted metabolomics based on ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) to reveal the core microbiota and metabolite profiles of pork during refrigeration. A total of 45 potential biomarkers were screened through random forest model analysis. Metabolic pathway analysis indicated that eleven pathways, including biogenic amine metabolism, pentose metabolism, purine metabolism, pyrimidine metabolism, phospholipid metabolism, and fatty acid degradation, were potential mechanisms of pork spoilage. Correlation analysis revealed nine metabolites-histamine, tyramine, tryptamine, D-gluconic acid, UDP-d-glucose, xanthine, glutamine, phosphatidylcholine, and hexadecanoic acid-as spoilage biomarkers, with Pseudomonas, Serratia, and Photobacterium playing significant roles. This study provides new insights into the changes in microbial and metabolic characteristics during the spoilage of refrigerated pork.

Insights into microbial contamination and antibiotic resistome traits in pork wholesale market: An evaluation of the disinfection effect of sodium hypochlorite.

Chlorine and its derivatives, such as sodium hypochlorite (NaClO) and chlorine dioxide, are frequently employed as disinfectants throughout the pork supply chain in China. Nevertheless, the extensive use of NaClO has the potential to cause the creation of 'chlorine-tolerant bacteria' and accelerate the evolution of antibiotic resistance. This study evaluated the efficacy of NaClO disinfection by examining alterations in the microbiome and resistome of a pork wholesale market (PWM), and bacteria isolation and analysis were performed to validate the findings. As expected, the taxonomic compositions of bacteria was significantly different before and after disinfection. Notably, Salmonella enterica (S. enterica), Salmonella bongori (S. bongori), Escherichia coli (E. coli), Klebsiella pneumoniae (K. pneumoniae), and Pseudomonas aeruginosa (P. aeruginosa) were observed on all surfaces, indicating that the application of NaClO disinfection treatment in PWM environments for pathogenic bacteria is limited. Correlations were identified between antibiotic resistance genes (ARGs) associated with aminoglycosides (aph(3'')-I, aph(6')-I), quinolone (qnrB, abaQ), polymyxin (arnA, mcr-4) and disinfectant resistance genes (emrA/BD, mdtA/B/C/E/F). Furthermore, correlations were found between risk Rank I ARGs associated with aminoglycoside (aph(3')-I), tetracycline (tetH), beta_lactam (TEM-171), and disinfectant resistance genes (mdtB/C/E/F, emrA, acrB, qacG). Importantly, we found that Acinetobacter and Salmonella were the main hosts of disinfectant resistance genes. The resistance mechanisms of the ARGs identified in PWM were dominated by antibiotic deactivation (38.7%), antibiotic efflux (27.2%), and antibiotic target protection (14.4%). The proportion of genes encoding efflux pumps in the PWM resistome increased after disinfection. Microbial cultures demonstrated that the traits of microbial contamination and antibiotic resistane were consistent with those observed by metagenomic sequencing. This study highlights the possibility of cross-resistance between NaClO disinfectants and antibiotics, which should not be ignored.

Co-exposure to sodium hypochlorite and cadmium induced locomotor behavior disorder by influencing neurotransmitter secretion and cardiac function in larval zebrafish.

Sodium hypochlorite (NaClO) and cadmium (Cd) are widely co-occurring in natural aquatic environment; however, no study has been conducted on effects of their combined exposure on aquatic organisms. To assess effects of exposure to NaClO and Cd in zebrafish larvae, we designed six treatment groups, as follows: control group, NaClO group (300 µg/L), 1/100 Cd group (48 µg/L), 1/30 Cd group (160 µg/L), NaClO+1/100 Cd group, and NaClO+1/30 Cd group analyzed behavior, neurological function and cardiac function. Results revealed that exposure to 1/30 Cd and NaClO+1/30 Cd caused abnormal embryonic development in larvae by altering body morphology and physiological indicators. Combined exposure to NaClO and 1/30 Cd affected the free-swimming activity and behavior of larvae in response to light-dark transition stimuli. Moreover, exposure to 1/30 Cd or NaClO+1/30 Cd resulted in a significant increase in tyrosine hydroxylase and acetylcholinesterase activities, as well as significant changes of various neurotransmitters. Lastly, exposure to 1/30 Cd or NaClO+1/30 Cd influenced the transcription of cardiac myosin-related genes and disturbed the myocardial contractile function. Altogether, our results suggested that combined exposure to NaClO and Cd induced oxidative damage in larvae, resulting in detrimental effects on nervous system and cardiac function, thus altering their swimming behavior.

Longitudinal metagenomic study reveals the dynamics of fecal antibiotic resistome in pigs throughout the lifetime.

BACKGROUND: The dissemination of antibiotic resistance genes (ARGs) poses a substantial threat to environmental safety and human health. Herein, we present a longitudinal paired study across the swine lifetime from birth to market, coupled with metagenomic sequencing to explore the dynamics of ARGs and their health risk in the swine fecal microbiome. RESULTS: We systematically characterized the composition and distribution of ARGs among the different growth stages. In total, 829 ARG subtypes belonging to 21 different ARG types were detected, in which tetracycline, aminoglycoside, and MLS were the most abundant types. Indeed, 134 core ARG subtypes were shared in all stages and displayed a growth stage-associated pattern. Furthermore, the correlation between ARGs, gut microbiota and mobile genetic elements (MGEs) revealed Escherichia coli represented the main carrier of ARGs. We also found that in most cases, the dominant ARGs could be transmitted to progeny piglets, suggesting the potential ARGs generation transmission. Finally, the evaluation of the antibiotic resistance threats provides us some early warning of those high health risk ARGs. CONCLUSIONS: Collectively, this relatively more comprehensive study provides a primary overview of ARG profile in swine microbiome across the lifetime and highlights the health risk and the intergenerational spread of ARGs in pig farm.

Differences in Bacterial Communities of Retail Raw Pork in Different Market Types in Hangzhou, China.

Pork is widely consumed globally, and pigs' microbiota can potentially harbor foodborne pathogens. Contaminated pork in retail markets poses significant implications for food quality and safety. However, limited studies have compared pork microbiomes in various marketing environments. In this study, we utilized traditional microbial culture methods and high-throughput 16S rRNA sequencing to assess pathogen contamination and bacterial diversity in raw pork samples purchased from farmers' markets and two types of supermarkets (upscale and ordinary) in Hangzhou, China. Traditional microbial plate cultures identified E. coli and Salmonella spp. in 32.1% (27/84) and 15.5% (13/84) of the collected pork samples, respectively. Moreover, 12 out of 13 Salmonella strains were found in farmers' markets. The MIC results indicated a high prevalence of MDR strains, accounting for 51.9% in E. coli and 53.8% in Salmonella. The prevalence of NaClO tolerant strains was 33.3% and 92.3% for E. coli and Salmonella, respectively. Sequencing results indicated significantly higher microbial diversity in farmers' market samples compared to supermarket samples. Farmers' market pork samples exhibited a greater abundance of Acinetobacter, while Pseudomonas and Brochothrix were predominant in supermarket samples. The total abundance of pathogenic and spoilage bacteria was also higher for the farmers' market samples. Cross-contamination during market trading was evident through a high correlation between bacterial abundance in pork from different stalls within the same farmers' market. PICRUSt2 analysis identified significant differences in the average proportions of genes for carbohydrate, energy, and lipid metabolism from the farmers' markets, suggesting an exacerbation of microbial metabolic activity and increased perishability of pork in this environment. In conclusion, this study revealed variations in the characteristics of raw pork bacterial contamination across different types of retail stores, as well as differences in the composition and diversity of their respective bacterial communities.

Aminobenzaldehyde convelently modified graphitic carbon nitride photocatalyst through Schiff base reaction: Regulating electronic structure and improving visible-light-driven photocatalytic activity for moxifloxacin degradation.

Graphitic carbon nitride (GCN) has been demonstrated to be a potential visible-light-driven photocatalyst for eliminating organic pollutants. However, its practical application is limited by low photocatalytic efficiency originating from high recombination of photogenerated charges. In this work, the electronic structure of GCN was regulated by doping aminobenzaldehyde (ABA) into the skeleton through a solid-state Schiff base reaction. Photoelectrochemical characterizations illustrated that the obtained catalyst (CNABA) exhibited narrower band gap, lower recombination rate of photoinduced electron-hole pair and higher charge transfer ability than the pristine GCN, further indicating its excellent photocatalytic activity. Using moxifloxacin (MOX) as a model pollutant, the ABA doping content was firstly optimized and the optimal activity of the CNABA photocatalyst was 3.1 times higher than that of the GCN. Subsequently, the effects of pH value, photocatalyst dosage and MOX concentration on the photocatalytic behavior were also studied. Superior stability and photocatalytic reusability were confirmed by five repeated tests. By substituting tap water/natural lake water for deionized water and sunlight for xenon light, the CNABA also exhibited high elimination efficiency, implying its good practicability. Superoxide radical, hydroxyl radical and photogenerated hole were identified as the main active species to degrade MOX. Finally, possible degradation pathways for MOX to mineralize small molecules were proposed by determining intermediate products during photocatalysis. This work provides a solid-state method to rationally design aromatic system-modified GCN photocatalysts, and the obtained CNABA is a promising visible-light-driven photocatalyst for eliminating antibiotics.

Dynamic Changes of Bacterial Communities and Microbial Association Networks in Ready-to-Eat Chicken Meat during Storage.

Ready-to-eat (RTE) chicken is a popular food in China, but its lack of food safety due to bacterial contamination remains a concern, and the dynamic changes of microbial association networks during storage are not fully understood. This study investigated the impact of storage time and temperature on bacterial compositions and microbial association networks in RTE chicken using 16S rDNA high-throughput sequencing. The results show that the predominant phyla present in all samples were Proteobacteria and Firmicutes, and the most abundant genera were Weissella, Pseudomonas and Proteus. Increased storage time and temperature decreased the richness and diversity of the microorganisms of the bacterial communities. Higher storage temperatures impacted the bacterial community composition more significantly. Microbial interaction analyses showed 22 positive and 6 negative interactions at 4 °C, 30 positive and 12 negative interactions at 8 °C and 44 positive and 45 negative interactions at 22 °C, indicating an increase in the complexity of interaction networks with an increase in the storage temperature. Enterobacter dominated the interactions during storage at 4 and 22 °C, and Pseudomonas did so at 22 °C. Moreover, interactions between pathogenic and/or spoilage bacteria, such as those between Pseudomonas fragi and Weissella viridescens, Enterobacter unclassified and Proteus unclassified, or those between Enterobacteriaceae unclassified and W.viridescens, were observed. This study provides insight into the process involved in RTE meat spoilage and can aid in improving the quality and safety of RTE meat products to reduce outbreaks of foodborne illness.

Genomic Investigation of Proteus mirabilis Isolates Recovered From Pig Farms in Zhejiang Province, China.

Proteus mirabilis is a common opportunistic zoonotic pathogen, and its ongoing acquisition of antimicrobial resistance genes poses challenges to clinical treatments. Human-sourced whole genomic sequencing of human P. mirabilis isolates has been reported, but pig-sourced isolates have not been thoroughly investigated even though these animals can serve as reservoirs for human infections. In the current study, we report a molecular epidemiological investigation to unravel the antimicrobial and virulence gene risk factors for P. mirabilis contamination in 9 pig farms in 3 different cities in Zhejiang Province, China. We collected 541 swab samples from healthy pigs and 30 were confirmed as P. mirabilis. All 30 isolates were resistant to tetracyclines, macrolides, sulfonamides, ß-lactams and chloramphenicol, and all were multiple drug-resistant and 27 were strong biofilm formers. Phylogenetic analyses indicated these 30 isolates clustered together in 2 major groups. Whole genome sequencing demonstrated that the isolates possessed 91 different antimicrobial resistance genes belonging to 30 antimicrobial classes including rmtB, sul1, qnrS1, AAC(6') - Ib - cr, blaCTX - M - 65 and blaOXA - 1. All isolates contained mobile genetic elements including integrative conjugative elements (ICEs) and integrative and mobilizable elements (IMEs). Minimum inhibitory concentration (MIC) testing indicated direct correlates between cognate genes and antimicrobial resistance. We also identified 95 virulence factors, almost all isolates contained 20 fimbrial and flagellar operons, and this represents the greatest number of these operon types found in a single species among all sequenced bacterial genomes. These genes regulate biofilm formation and represent a confounding variable for treating P. mirabilis infections. Our P. mirabilis isolates were present in healthy animals, and multiple drug resistance in these isolates may serve as a reservoir for other intestinal and environmental Enterobacteriaceae members. This prompts us to more strictly regulate veterinary antibiotic use.

Transcriptomic Responses of Salmonella enterica Serovars Enteritidis in Sodium Hypochlorite.

Salmonella enterica serovars Enteritidis (S. Enteritidis) can survive extreme food processing environments including bactericidal sodium hypochlorite (NaClO) treatments generally recognized as safe. In order to reveal the molecular regulatory mechanisms underlying the phenotypes, the overall regulation of genes at the transcription level in S. Enteritidis after NaClO stimulation were investigated by RNA-sequencing. We identified 1399 differentially expressed genes (DEG) of S. Enteritidis strain CVCC 1806 following treatment in liquid culture with 100 mg/L NaClO for 20 min (915 upregulated and 484 downregulated). NaClO stress affects the transcription of genes related to a range of important biomolecular processes such as membrane damage, membrane transport function, energy metabolism, oxidative stress, DNA repair, and other important processes in Salmonella enterica. First, NaClO affects the structural stability of cell membranes, which induces the expression of a range of outer and inner membrane proteins. This may lead to changes in cell membrane permeability, accelerating the frequency of DNA conversion and contributing to the production of drug-resistant bacteria. In addition, the expression of exocytosis pump genes (emrB, yceE, ydhE, and ydhC) was able to expel NaClO from the cell, thereby increasing bacterial tolerance to NaClO. Secondly, downregulation of genes related to the Kdp-ATPase transporter system (kdpABC) and the amino acid transporter system (aroP, brnQ and livF) may to some extent reduce active transport by bacterial cells, thereby reducing their own metabolism and the entry of disinfectants. Downregulation of genes related to the tricarboxylic acid (TCA) cycle may drive bacterial cells into a viable but non-culturable (VBNC) state, resisting NaClO attack by reducing energy metabolism. In addition, significant upregulation of genes related to oxidative stress could mitigate damage caused by disinfectants by eliminating alkyl hydroperoxides, while upregulation of genes related to DNA repair could repair damage to bacterial cells caused by oxidative stress. Therefore, this study indicated that S. Enteritidis has genomic mechanisms to adapt to NaClO stress.

Prevalence and characteristics of the mcr-1 gene in retail meat samples in Zhejiang Province, China.

Considering the serious threat to food safety and public health posed by pathogens with colistin resistance, colistin was banned as a growth promoter in 2017 in China. In recent years, the resistance rate of Escherichia coli isolated from animal intestines or feces to colistin has decreased. However, the prevalence and characteristics of the mcr-1 gene in retail meat have not been well explored. Herein, 106 mcr-1-negative and 16 mcr-1-positive E. coli isolates were randomly recovered from 120 retail meat samples and screened using colistin. The 106 E. coli isolates showed maximum resistance to sulfafurazole (73.58%) and tetracycline (62.26%) but susceptibility to colistin (0.00%). All 16 mcr-1-positive E. coli isolates showed resistance to colistin, were extended spectrum beta-lactamase (ESBL)-positive and exhibited complex multidrug resistance (MDR). For these 16 isolates, 17 plasmid replicons and 42 antibiotic resistance genes were identified, and at least 7 antibiotic resistance genes were found in each isolate. Acquired disinfectant resistance genes were identified in 75.00% (12/16) of the isolates. Furthermore, comparative genomic and phylogenetic analysis results indicated that these 16 mcr-1-positive E. coli isolates and the most prevalent mcr-1-harboring IncI2 plasmid in this study were closely related to other previously reported mcr-1-positive E. coli isolates and the IncI2 plasmid, respectively, showing their wide distribution. Taken together, our findings showed that retail meat products were a crucial reservoir of mcr-1 during the colistin ban period and should be continuously monitored.

Escherichia fergusonii, an Underrated Repository for Antimicrobial Resistance in Food Animals.

A total of 1,400 samples of food animals (pigs, chickens, and ducks) were collected between July and September 2019 in China to uncover the prevalence of E. fergusonii and its potential role in the evolution of antimicrobial resistance (AMR). An isolation of E. fergusonii was performed and pulsed-field gel electrophoresis (PFGE) was used to uncover the genetic relationship. The AMR of E. fergusonii isolates was comprehensively characterized using broth microdilution-based antimicrobial susceptibility testing, S1-PFGE, southern hybridization, whole-genome sequencing, and in-depth bioinformatics analysis. As a result, a total of 133 E. fergusonii isolates were obtained. These isolates could be grouped into 41 PFGE subclades, suggesting a diverse genetic relationship. The resistance phenotypes of sulfafurazole (97.74%) and tetracycline (94.74%) were the most frequently found. Of the E. fergusonii isolates, 51.88% were extended spectrum beta-lactamase (ESBL)-positive. Forty-three different AMR genes were revealed based on 25 genome sequences harboring mcr-1. Briefly, aph(6)-Id, aph(3'')-Ib and tet(A) genes were the most frequently observed, with the highest rate being 76.00% (19/25). Three mcr-1-harboring plasmids were identified after Nanopore sequencing, including pTB31P1 (IncHI2-IncHI2A, 184,652 bp), pTB44P3 (IncI2, 62,882 bp), and pTB91P1 (IncHI2-IncHI2A, 255,882 bp). Additionally, 25 E. fergusonii isolates harboring mcr-1 were clustered together with other E. fergusonii isolates from different regions and sources available in GenBank, suggesting a possible random process of mcr-1 transmission in E. fergusonii. In conclusion, E. fergusonii is widespread in food animals in China and might be an important reservoir of AMR genes, especially mcr-1, and facilitate the evolution of AMR. IMPORTANCEE. fergusonii, a member of the genus Escherichia, has been reported to transmit via the food chain and cause diseases in humans. However, the prevalence of multidrug-resistant E. fergusonii, especially mcr-1-positive E. fergusonii isolates, has rarely been reported. Here, we collected 1,400 samples from food animals in three provinces of China and obtained 133 E. fergusonii isolates (9.5%). We found that the prevalence of E. fergusonii isolates was diverse, with high levels of antimicrobial resistance. Among them, 18.8% E. fergusonii isolates carried the colistin resistance gene mcr-1. Thus, E. fergusonii may facilitate the evolution of colistin resistance as a reservoir of mcr-1. As far as we know, the prevalence and AMR of E. fergusonii in the food animals in this study was first reported in China. These findings increase our understanding of the role of E. fergusonii in public health and the evolution of antibiotic resistance.

Evaluation of the Stress Tolerance of Salmonella with Different Antibiotic Resistance Profiles.

Disease caused by antibiotic-resistant Salmonella is a serious clinical problem that poses a great threat to public health. The present study is aimed at assessing differences in bacterial kinetics with different antibiotic resistance profiles under environmental stress and at developing microbial tolerance models in lettuce during storage from 4 to 36°C. The drug-resistance phenotypes of 10 Salmonella Typhimurium (S. Typhimurium) isolates were examined using the broth microdilution method. The results of 10 S. Typhimurium isolates in the suspensions showed that a slow trend towards reduction of drug-sensitive (DS) isolates in relation to the others though without statistical difference. Compared to DS S. Typhimurium SA62, greater bacterial reduction was observed in multidrug-resistant (MDR) S. Typhimurium HZC3 during lettuce storage at 4°C (P < 0.05). It was likely that a cross-response between antibiotic resistance and food-associated stress tolerance. The greater growth in lettuce at 12°C was observed for DS S. Typhimurium SA62 compared to MDR S. Typhimurium HZC3 and was even statistically different (P < 0.05), while no significant difference was observed for bacterial growth between MDR S. Typhimurium HZC3 and DS S. Typhimurium SA62 strains in lettuce storage from 16 to 36°C (P > 0.05). The goodness-of-fit indices indicated the Log-linear primary model provided a satisfactory fit to describe the MDR S. Typhimurium HZC3 and DS S. Typhimurium SA62 survival at 4°C. A square root secondary model could be used to describe the effect of temperature (12, 16, 28, and 36°C) on the growth rates of S. Typhimurium HZC3 (adj - R 2 = 0.91, RMSE = 0.06) and S. Typhimurium SA62 (adj - R 2 = 0.99, RMSE = 0.01) derived from the Huang primary model. It was necessary to pay attention to the tolerance of antibiotic resistant bacteria under environmental stress, and the generated models could provide parts of the input data for microbial risk assessment of Salmonella with different antibiotic resistance profile in lettuce.

Modulation of the Intestinal Microbiota by the Early Intervention with Clostridium Butyricum in Muscovy Ducks.

This study evaluated the effects of early intervention with Clostridium butyricum (C. butyricum) on shaping the intestinal microbiota of Muscovy ducklings. A total of 160 1-day-old male ducks were randomly divided into two groups: the CB group was administered with 1 mL of C. butyricum (2 × 109 CFU/mL), while the C group was given 1 mL of saline. The administration lasted for 3 days. We found that C. butyricum had no significant effect on growth performance. The results indicated that inoculation with C. butyricum could significantly increase the abundance of genera Bacteroides, Lachnospiraceae_uncultured, and Ruminococcaceae on Day 14 and reduce the abundance of Escherichia-Shigella and Klebsiella on Days 1 and 3. Moreover, the CB group ducks had higher concentrations of acetic, propionic, and butyrate in the cecum than the C group. Overall, these results suggest that early intervention with C. butyricum could have positive effects on Muscovy ducks' intestinal health, which might be attributed to the modulation in the intestinal microbial composition and the increased concentrations of short-chain fatty acids (SCFAs). C. butyricum might even have the potential to help the colonization of beneficial bacteria in the intestine microbiota in Muscovy ducks in poultry and other livestock.

Chlorine Tolerance and Cross-Resistance to Antibiotics in Poultry-Associated Salmonella Isolates in China.

Chlorine disinfectants have been widely used in the poultry supply chain but this exposure can also result in the development of bacterial tolerance to chlorine and this is often linked to antibiotic cross-resistance. The objectives of this study were to investigate sodium hypochlorite (NaClO) tolerance of Salmonella isolated from poultry supply chains and evaluate cross-resistance. We collected 172 Salmonella isolates from poultry farms, slaughter houses and retail markets in China during 2019-2020. We found that S. Enteritidis, S. Kentucky, and S. Typhimurium constituted > 80% of our Salmonella isolates. Overall, 68% of Salmonella isolates were resistant to > 3 antibiotics and S. Kentucky displayed a significantly (p > 0.05) higher frequency (93.2%) of multidrug resistance than the other serovars. Tolerance to chlorine at MIC > 256 mg/L was detected in 93.6% of isolates (161/172) and tolerant isolates displayed higher decimal reduction times (D value) and less ultrastructural damage than did the suspectable strains under chlorine stress. Spearman analysis indicated significant positive correlations between chlorine tolerance (evaluated by the OD method) and antibiotic resistance (p < 0.05) to ceftiofur, tetracycline, ciprofloxacin and florfenicol and this was most likely due to efflux pump over-expression. The most frequently detected chlorine resistance gene was qacEΔ1 (83.1%, n = 143) and we found a positive correlation between its presence and MIC levels (r = 0.66, p < 0.0001). Besides, we found weak correlations between chlorine-tolerance and antibiotic resistance genes. Our study indicated that chlorine disinfectants most likely played an important role in the emergence of chlorine tolerance and spread of antibiotic resistance and therefore does not completely control the risk of food-borne disease. The issue of disinfectant resistance should be examined in more detail at the level of the poultry production chain.

Modeling the Reduction and Cross-Contamination of Salmonella in Poultry Chilling Process in China.

: The study was to establish a predictive model for reduction and cross-contamination of Salmonella on chicken in chilling process. Reduction of Salmonella on chicken was 0.75 ± 0.04, 0.74 ± 0.08, and 0.79 ± 0.07 log CFU/g with 20, 50, and 100 mg/L of chlorine, respectively. No significant differences of bacterial reductions with 20-100 mg/L of chlorine were found and a Normal (-0.75, 0.1) distribution could describe the uncertainty of bacterial reductions. Inoculated and non-inoculated chicken samples were washed together and bacterial transfer rates among them were 0.13%-0.004% with 20-100 mg/L of chlorine. No significant differences of transfer rates with 50-100 mg/L of chlorine were observed and a Triangle (-2.5, -1.5, -1.1) distribution could describe the log transfer rate. Additionally, a 3-factor response surface model based on the central composite design was developed to evaluate the effects of initial contamination level (1-5 log CFU/g), pre-chill incidence (3%-40%) and chlorine concentration (0-100 mg/L) on post-chill incidence. The post-chill incidences in these treatments were within 30%-91.7%. The developed model showed a satisfactory performance to predict the post-chill incidence as evidenced by statistical indices (pseudo-R2 = 0.9; p < 0.0001; RMSE = 0.21) and external validation parameters (Bf = 1.02; Af = 1.11).

Modeling the Reduction of Salmonella spp. on Chicken Breasts and Wingettes during Scalding for QMRA of the Poultry Supply Chain in China.

The objective of this study was to develop predictive models for describing the inoculated Salmonella reductions on chicken during the scalding process in China. Salmonella reductions on chicken breasts at a 100 s treatment were 1.12 ± 0.07, 1.38 ± 0.01, and 2.17 ± 0.11 log CFU/g at scalding temperatures of 50, 60 and 70 °C, respectively. For chicken wingettes, 0.87 ± 0.02, 0.99 ± 0.14 and 1.11 ± 0.17 log CFU/g reductions were obtained at 50, 60 and 70 °C after the 100 s treatment, respectively. Greater bacterial reductions were observed on chicken breasts than on chicken wingettes (p < 0.05). A logistic (-1.12, 0.06) distribution could describe the bacterial reductions on chicken breasts at 50-60 °C. Weibull, exponential and log-linear models were compared for describing the bacterial reduction on chicken breasts at 70 °C and the Weibull model showed the best fit as indicated by the pseudo-R2, root mean square error (RMSE) and standard error of prediction (SEP) values. For chicken wingettes, a logistic (-0.95, 0.07) distribution could be used to describe the bacterial reduction at 50-70 °C. The developed predictive models could provide parts of the input data for microbial risk assessment of the poultry supply chain in China.

Behavior of Salmonella Typhimurium on Fresh Strawberries Under Different Storage Temperatures and Wash Treatments.

Fresh strawberries are one of the most popular fruits in China and are vulnerable to microbial contamination. In this study, the behavior of Salmonella Typhimurium on fresh strawberries stored at refrigeration and room temperatures, as well as the effectiveness of mild heat wash treatments at 47, 50, and 53°C on bacterial survival was investigated. The modified Gompertz, Huang, log-linear, and Weibull models were used to fit bacterial growth and survival curves under different treatments. A secondary model based on linear regression was developed to describe the effect of washing temperature on the kinetic parameters of S. Typhimurium survival derived from the Weibull model. During 72 h storage, S. Typhimurium on fresh strawberries stored at 4°C was reduced by 1.35 log CFU/g and growth of 5.64 log CFU/g was observed when strawberries were stored at 25°C. Bacterial reductions of 1.22 ± 0.15, 1.92 ± 0.06, 2.27 ± 0.07 log CFU/g were obtained when washing was carried out at 47, 50 and 53°C for 240 s, respectively. The wash temperature was an important parameter for bacterial inactivation and bacterial populations declined significantly in conjunction with washing time (p < 0.05). Warm wash treatments lead the visible color changes of strawberries, showing a slightly darker appearance while acceptable. The goodness-of-fit indices indicated that the log-linear model provided a satisfactory fit to describe the bacterial survival at 4°C. According to the smaller Akaike information criterion (AIC) value, the modified Gompertz model performed slightly better than the Huang model in describing bacterial growth at 25°C. The high adj-R2 (≥0.90) and small RMSE (≤0.22) indicated the Weibull model better described bacterial behavior under mild heat treatments. We found a close linear relationship between wash temperatures and ln k and ln n. These models were validated by independent experimental data and the values of the bias and accuracy factors fell into the acceptable range.

Modeling Transfer of Vibrio Parahaemolyticus During Peeling of Raw Shrimp.

This study aimed to qualify the transfer of Vibrio parahaemolyticus during the shrimp peeling process via gloves under 3 different scenarios. The 1st 2 scenarios provided quantitative information for the probability distribution of bacterial transfer rates from (i) contaminated shrimp (6 log CFU/g) to non-contaminated gloves (Scenario 1) and (ii) contaminated gloves (6 log CFU/per pair) to non-contaminated shrimp (Scenario 2). In Scenario 3, bacterial transfer from contaminated shrimp to non-contaminated shrimp in the shrimp peeling process via gloves was investigated to develop a predictive model for describing the successive bacterial transfer. The range of bacterial transfer rate (%) in Scenarios 1 and 2 was 7% to 91.95% and 0.04% to 12.87%, respectively, indicating that the bacteria can be transferred from shrimp to gloves much easier than that from gloves to shrimp. A Logistic (1.59, 0.14) and Triangle distribution (-1.61, 0.12, 1.32) could be used to describe the bacterial transfer rate in Scenarios 1 and 2, respectively. In Scenario 3, a continuously decay patterning with fluctuations as the peeling progressed has been observed at all inoculation levels of the 1st shrimp (5, 6, and 7 log CFU/g). The bacteria could be transferred easier at 1st few peels, and the decreasing bacterial transfer was found in later phase. Two models (exponential and Weibull) could describe the successive bacterial transfer satisfactorily (pseudo-R2 > 0.84, RMSE < 1.23, SEP < 10.37). The result of this study can provide information regarding cross-contamination events in the seafood factory. PRACTICAL APPLICATION: This study presented that Vibrio parahaemolyticus cross-contamination could be caused by gloves during the shrimp peeling process. The bacterial transfer rate distribution and predictive model derived from this work could be used in risk assessment of V. parahaemolyticus to ensure peeled shrimp safety.