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, β-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.

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.

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 (B_f = 1.02; A_f = 1.11).

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.

Viable but non-culturable (VBNC) bacteria after chlorination disinfection pose a considerable threat to the public health and environment. The current work aimed to uncover the stress adaptation mechanisms of VBNC S. Enteritidis to sodium hypochlorite. In this study, we investigated the morphology, gene expressions and proteomic profile in VBNC S. Enteritidis. It demonstrated that 10⁹ cells mL⁻¹ of bacteria could transition into the VBNC state after exposure to 50 mg L⁻¹ chlorine for 2.5 h. The VBNC state cells exhibited cellular membrane impairment, decreased size, and intracellular ATP depletion compared to the culturable cells. In the VBNC state cells, there was a significant increase in intracellular reactive oxygen species (ROS) levels and NAD+/NADH ratios. This observation suggested an imbalance in the intracellular redox status of the VBNC bacteria. Transcriptome and proteomic analysis revealed significant upregulation of the resistance-nodulation-division (RND) efflux systems (arcA, tolC, ramA), oxidative stress response genes (soxR, trxA, grxB), DNA repair genes (lexA, radA, mutH, mutL), and virulence factor genes (sseA, sseL, eutS, and eutN). Our findings indicated that it is important to formulate disinfection protocols that are specifically designed for food processing plants and healthcare facilities, focusing on implementing a more thorough disinfection method to effectively eradicate VBNC cells.

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.