Tigecycline is a last-resort antibiotic for treating infections caused by carbapenem-resistant Enterobacterales (CRE). The emergence and dissemination of tigecycline-resistant Enterobacterales through the food chain pose a growing public health concern. This study systematically evaluates the global prevalence of tigecycline-resistant Enterobacterales and the tet(X4) carriage rate in foods and food-producing animals, and analyzes the epidemiological characteristics across supply chain stages, bacterial genera, regions, and sample types. We searched PubMed, Scopus, Web of Science, Engineering Index (EI), and China National Knowledge Infrastructure (CNKI) for studies published from January 2015 to December 2025. A random-effects model pooled resistance rates, and subgroup analyses explored heterogeneity. A total of 104 studies (130 reports) were included. The overall resistance rate was 4.36% (95% CI: 2.83%-6.20%); among resistant strains, tet(X4) prevalence (13 reports) was 73.81% (95% CI: 43.79%-94.96%). E.coli accounted for 93.01% of resistant strains. The highest resistance rate was at farms (5.05%), followed by retail markets (2.61%) and fruits and vegetables (3.82%). Asia showed a significantly higher rate (5.30%) than Europe (2.68%). Although overall prevalence is low, tet(X4) is highly prevalent among resistant strains, with E. coli as the predominant host. Farms and Asia are key control points, and resistance in fruits and vegetables warrants vigilance. Strengthening veterinary tetracycline regulation and incorporating tigecycline resistance monitoring into routine food chain surveillance are strongly recommended.

Global bacterial antibiotic resistance threatens health, food safety, and sustainability. The food supply chain is a critical "One Health" pathway, linking agriculture, environment, and processing. However, systematic reviews addressing the impact of coexisting stressors on antibiotic resistance emergence and transmission across this continuum are lacking. This review innovatively synthesizes environmental inputs (antibiotic residues, fertilizers, heavy metals, pesticides, microplastics, climate change, and grazing) and processing/transport stressors (temperature, nonthermal technologies, pH, osmosis, disinfectants, food additives, probiotics, and trade), focusing on their individual and synergistic effects. These stressors enhance resistance and horizontal gene transfer by activating bacterial stress responses (sigma factors, SOS), altering membranes, and triggering mutations/efflux pumps. Coexisting stressors can further intensify, accelerate, and amplify resistance emergence and transmission. We propose multilevel mitigation strategies across the food chain, including curbing selective pressures at the source, optimizing food processing techniques to avoid stress-induced resistance, guiding consumer behavior, and strengthening international regulatory governance.

ObjectiveTo understand the influence of cultivation conditions on the formation of Salmonella biofilm and provide basis for ensure food safety.MethodsCongo red staining and crystal violet staining method were used to screen biofilm （BF） formation strains from 400 foodborne Salmonella. Scanning electron microscope was applied to observe BF formation. Whole-genome sequencing analysis was used to identify BF formation related genes. Tube culture and microplate quantitative detection methods were used to determine BF formation of Salmonella under different culture conditions. The effect of different culture time， medium type， glucose content， peptone content and Vitamin B1 （VB1） content to Salmonella BF formation were studied.ResultsEight （2.00%） BF-forming Salmonella isolates were obtained. Compared to LB medium， BHI was more suitable for BF formation for Salmonella. When cultured for 96 h， the BF formation of Salmonella on BHI was much more than that on LB. Fifteen BF-related genes including trx2， fadI， bsmA， bssS， tabA， bcsE， csgD， csgA， B， C， seqA， dam， fliZ， flhD and motB were detected in BJ44D， ZZSR2-2008， NYSR94-08 and yl19Ta. The best BF formation nutrient conditions for Salmonella were that 0.6% or 0.8% glucose， 4.0% peptone， and 50 mg/L VB1 were respectively supplemented in BHI broth （P<0.05）.ConclusionIt is indicated that different culture conditions significantly correlated with BF formation of Salmonella and have strain-specificity， and can provide theoretical basis for BF formation control of Salmonella in the food production chain.

With the continuous advancements in high-throughput genome sequencing technologies and the development of innovative bioinformatics tools, bacterial genome-wide association studies (BGWAS) have emerged as a transformative approach for investigating the genetic variations underlying diverse bacterial phenotypes at the population genome level. This review provides a comprehensive overview of the application of BGWAS in elucidating genetic determinants of bacterial drug resistance, pathogenicity, host specificity, biofilm formation, and probiotic fermentation characteristics. We systematically summarize the BGWAS workflow, including study design, data analysis pipelines, and the bioinformatics software employed at various stages. Furthermore, we highlight specialized tools tailored for BGWAS and discuss their unique features and applications. We also discuss confounding factors that can influence the accuracy and reliability of BGWAS results, including population structure, linkage disequilibrium, and multiple testing. By incorporating recent advancements, this review serves as a comprehensive reference for researchers utilizing BGWAS to uncover the genetic basis of bacterial phenotypes.

Cronobacter spp. exhibit remarkable resilience to extreme environmental stresses, including thermal, acidic, desiccation, and osmotic conditions, posing significant challenges to food safety. Their thermotolerance relies on heat shock proteins (HSPs), thermotolerance genomic islands, enhanced DNA repair mechanisms, and metabolic adjustments, ensuring survival under high-temperature conditions. Acid tolerance is achieved through internal pH regulation, acid efflux pumps, and acid tolerance proteins, allowing survival in acidic food matrices and the gastrointestinal tract. Desiccation tolerance is mediated by the accumulation of protective osmolytes like trehalose, stabilizing proteins and membranes to withstand dryness, especially in dry food products. Similarly, osmotic stress resilience is supported by compatible solutes such as trehalose and glycine betaine, along with metabolic adaptations to balance osmotic pressures. These mechanisms highlight the adaptability of Cronobacter spp. to diverse environments. Moreover, exposure to sublethal stresses, including heat, osmotic, dry, and pH stresses, may induce homologous or cross-resistance, complicating control strategies. Understanding these survival mechanisms is essential to mitigate the risks of Cronobacter spp., especially in powdered infant formula (PIF), and ensure food safety.

As emerging environmental pollutants, antibiotic resistance genes (ARGs) are prevalent in livestock farms and their surrounding environments. Although existing studies have focused on ARGs in specific environmental media, comprehensive research on ARGs within farming environments and their adjacent areas remains scarce. This review explores the sources, pollution status, and transmission pathways of ARGs from farms to the surrounding environment. Drawing on the "One Health" concept, it also discusses the potential risks of ARGs transmission from animals to human pathogens and the resulting impact on human health. Our findings suggest that the emergence of ARGs in livestock farming environments primarily results from intrinsic resistance and genetic mutations, while their spread is largely driven by horizontal gene transfer. The distribution of ARGs varies according to the type of resistance genes, seasonal changes, and the medium in which they are present. ARGs are disseminated into the surrounding environment via pathways such as manure application, wastewater discharge, and aerosol diffusion. They may be absorbed by humans, accumulating in the intestinal microbiota and subsequently affecting human health. The spread of ARGs is influenced by the interplay of microbial communities, antibiotics, heavy metals, emerging pollutants, and environmental factors. Additionally, we have outlined three control strategies: reducing the emergence of ARGs at the source, controlling their spread, and minimizing human exposure. This article provides a theoretical framework and scientific guidance for understanding the cross-media migration of microbial resistance in livestock farming environments.

This research presents a comprehensive review of Salmonella presence in retail fresh fruits and vegetables from 2010 to 2023, utilizing data from recognized sources such as PubMed, Scopus, and Web of Science. The study incorporates a meta-analysis of prevalence, serovar distribution, antimicrobial susceptibility, and antimicrobial resistance genes (ARGs). Additionally, it scrutinizes the heterogeneous sources across various food categories and geographical regions The findings show a pooled prevalence of 2.90% (95% CI: 0.0180-0.0430), with an increase from 4.63% in 2010 to 5.32% in 2022. Dominant serovars include S. Typhimurium (29.14%, 95% CI: 0.0202-0.6571) and S. Enteritidis (21.06%, 95% CI: 0.0181-0.4872). High resistance rates were noted for antimicrobials like erythromycin (60.70%, 95% CI: 0.0000-1.0000) and amoxicillin (39.92%, 95% CI: 0.0589-0.8020). The most prevalent ARGs were blaTEM (80.23%, 95% CI: 0.5736-0.9692) and parC mutation (66.67%, 95% CI: 0.3213-0.9429). Factors such as pH, water activity, and nutrient content, along with external factors like the quality of irrigation water and prevailing climatic conditions, have significant implications on Salmonella contamination. Nonthermal sterilization technologies, encompassing chlorine dioxide, ozone, and ultraviolet light, are emphasized as efficacious measures to control Salmonella. This review stresses the imperative need to bolster prevention strategies and control measures against Salmonella in retail fresh fruits and vegetables to alleviate related food safety risks.

Cronobacter sakazakii is an opportunistic pathogen known for causing severe diseases. Mild heat treatment is commonly used in food processing, however, the pathogenic characteristics and underlying mechanisms of Cronobacter sakazakii strains remain poorly understood. In this study, we found that mild heat stress (MHS) at 52 degrees C can induce several deleterious effects in Cronobacter sakazakii, including damage to the cell wall, genomic DNA breakage, and misfolding of cytoplasmic proteins. These conditions lead to a decreased survival ability under acid, desiccation, and osmotic stress; a reduction in biofilm formation; and diminished motility. Notably, surviving C. sakazakii cells retain their pathogenicity, causing significant intestinal damage in newborn mice. This damage is characterized by epithelial sloughing and disruption of the intestinal structure. Tandem mass tag (TMT)-based proteomics identified 736 proteins with differential abundance across C. sakazakii strains subjected to mild heat stress, highlighting adaptations in biofilm formation, motility, and stress tolerance. Key regulatory changes were observed in phospholipid metabolism and protein synthesis, which underpin this complex stress response. This data illustrates a sophisticated balance between environmental adaptability and pathogenic potential. The metabolic and pathogenic responses of C. sakazakii to mild heat stress are closely linked to its phospholipid metabolism and the production of secretory proteins, both crucial for its virulence and reliant on membrane transport. This complex interplay emphasizes the need to understand these mechanisms to develop effective control strategies.

This experiment established a relative quantitative detection method for bovine derived components in milk powder based on Real time PCR.The specificity,sensitivity,and stability of cattle-specific primers and probes were tested.By simulating mixed samples of cow's milk powder and horse's milk powder with different concentrations,linear fitting was performed based on the functional relationship of their △Ct values,and a standard curve was drawn to establish a relative quantitative detection of bovine derived components in milk powder.The results showed that the minimum detection limit of this method was 0.000 01 mg/mL.The recovery rate was 91.11%～119.2%,with an inter group coefficient of variation of≤0.58%and an intra group coefficient of variation of≤1.44%.This method is suitable for detecting the adulteration of bovine derived components and content in milk powder in terms of specificity and stability.

Abstract This project presents a quantitative detection method to identify raccoon‐derived ingredient adulteration in sausage products. The specific copy gene of the raccoon was selected as the target gene. According to the specificity of its primer and probe, the quantitative detection method of raccoon microdrops by droplet digital PCR was established. In addition, the accuracy of the proposed method was verified by artificially mixed samples, and the applicability of this method was tested based on the commercially available products. The experimental results indicate that the raccoon mass (M) and raccoon‐extracted DNA concentration have a good linear relationship when the sample content is 5–100 mg, and there is also a significant linear relationship between DNA content and DNA copy number (C) with R2 = .9982. Therefore, using DNA concentration as the median signal, the conversion equation between raw raccoon mass (M) and DNA copy number (C) could be obtained as follows: M = (C + 177.403)/16.954. The detection of artificially mixed samples and commercial samples shows that the method is accurate and suitable for quantitative adulteration detection of various sausage products in the market.

Global occurrences of foodborne disease outbreaks have been documented, involving fresh agricultural produce contaminated by various pathogens. This contamination can occur at any point in the supply chain. However, studies on the prevalence of total coliforms, Salmonella and microbial diversity in vegetable and associated environments are limited. This study aimed to assess 1) the number of total coliforms (n = 299) and diversity of microbial communities (n = 52); 2) the prevalence, antibiotic susceptibility, genomic characteristics, and potential transmission relationships of Salmonella in soil-irrigation water-vegetable system (n = 506). Overall, 84.28 % samples were positive to total coliforms, with most frequently detected in soil (100 %), followed by irrigation water (79.26 %) and vegetables (62.00 %). A seasonal trend in coliform prevalence was observed, with significantly higher levels in summer (P < 0.05). Detection rates of Salmonella in soil, vegetable and irrigation water were 2.21 %, 4.74 % and 9.40 %. Fourteen serotypes and sequence types (STs) were respectively annotated in 56 Salmonella isolates, ST13 S. Agona (30.36 %, 17/56), ST469 S. Rissen (25.00 %, 14/56), and ST36 S. Typhimurium (12.50 %, 7/56) were dominant serotypes and STs. Thirty-one (55.36 %) isolates were multi-drug resistant, and the resistance was most frequently found to ampicillin (55.36 %, 31/56), followed by to sulfamethoxazole (51.79 %, 29/56) and tetracycline (50.00 %, 28/56). The genomic characteristics and antibiotic resistance patterns of Salmonella isolates from soil, vegetables, and irrigation water within a coherent geographical locale exhibited remarkable similarities, indicating Salmonella may be transmitted among these environments or have a common source of contamination. Microbial alpha diversity indices in soil were significantly higher (P < 0.05) than that in vegetable and irrigation water. The microbial phylum in irrigation water covered that in the vegetable, demonstrating a significant overlap in the microbial communities between the vegetables and the irrigation water.

Salmonella exhibits extensive genetic diversity, facilitated by horizontal gene transfer occurring within and between species, playing a pivotal role in this diversification. Nevertheless, most studies focus on clinical and farm animal isolates, and research on the pangenome dynamics of Salmonella isolates from retail stage of the animal food supply chain is limited. Here, we investigated the genomes of 950 Salmonella isolates recovered from retail chicken and pork meats in seven provinces and one municipality of China in 2018. We observed a strong correlation between Salmonella sublineage diversity and the accessory genome with meat type, revealing reduced diversity associated with increased resistance. Importantly, genes associated with antibiotic, biocide, and heavy metal resistance were unevenly distributed in Salmonella from retail chicken and pork. Pork Salmonella isolates showed a higher prevalence of copper and silver resistance genes, while chicken Salmonella isolates displayed a significant predominance of genetic determinants associated with cephalosporin and ciprofloxacin resistance. Moreover, co-occurrence patterns of resistance determinants and their interaction with mobile genetic elements also correlated with meat type. In summary, our findings shed light on how Salmonella achieves their ecological niche success driven by evolution and gene changes in the retail stage of the animal food supply chain.

The authentication of dairy species has great significance for food safety. This study focused on a more rapid method for identifying major dairy species, and specific recombinase polymerase amplification (RPA)-based assays for cattle, goat, sheep, camel and donkey were developed. Through the developed RPA-based assays, goats and sheep could be simultaneously identified and bovine families could be differentiated. The performances of the RPA assays were validated using 37 milk powder samples, of which 16.2% (6/37) were suspected of being adulterated and 24.3% (9/37) were potentially at risk of being wrongly identified as adulteration. The effectiveness of the developed assays for crude DNA detection was also validated by a rapid nucleic acid extraction kit, and results showed that the presence of large amounts of protein and fat did not affect the qualitative results. Therefore, these assays could combine with the rapid nucleic acids extraction methods for being used in field detection.

In this study, four lactic acid bacteria (LAB) strains demonstrating ciprofloxacin, bile salt, gastric fluid, and intestinal fluid tolerance as well as adhesion ability to Caco-2 and HT-29 cells were used to improve and recover the intestinal flora disorders caused by ciprofloxacin, among which, Lactobacillus brevis 505 exhibited excellent adhesion ability to two kinds of cells and colonization ability to mouse intestinal. After ciprofloxacin treatment, certain recovery effect on cecum caused by ciprofloxacin in the mice was found during natural recovery (group 5C2), but it was challenging to fully restore the intestinal integrity to the initial level. After L. brevis 505 intervention (group 5C5), the intestinal damage to the colon and ileum caused by ciprofloxacin in mice was significantly alleviated; the recovery effect was better than that of natural recovery. Additionally, L. brevis 505 could effectively regulate INF-γ, sIgA, and RegIIIγ increase induced by ciprofloxacin. Shannon and Simpson index of the intestinal flora of mice in 5C5 group were higher than those in other group, the relative abundance of Bifidobacterium and Lactobacillus in the mice in 5C5 group was increased, indicating that LAB can better restore the structure and abundance of intestinal microflora. Consequently, L. brevis 505 shows promise as a probiotic for gut microbiota restoration and rebuilding during antibiotic therapy.

This paper reported a novel approach to quantification of adulterated fox-derived components in meat products by drop digital polymerase chain reaction (ddPCR). By using the F2 gene as the target gene of fox, a single primer was designed to identify the adulteration that had been added either inadvertently or deliberately during the process. In this paper, the fox meat was used as the experimental materials and a relationship was established between fox mass and copy number by extracting DNA and using DNA concentration as an intermediary. The results that across the dynamic range, the relationships between meat mass and DNA concentration were nearly linear (R2 = 0.9986), as was the relationship between DNA concentration and DNA copy number (R2 = 0.9992). Based on the DNA concentration, the following formulas were developed about the relationship between fox meat mass (Mfox) and DNA copy number (C): Mfox = 0.05C + 2.7.

The rise of antibiotic resistance in Escherichia coli has become a major global public health concern. While there is extensive research on antibiotic-resistant E. coli from human and animal sources, studies on vegetables and their environments are limited. This study investigated the prevalence and characteristics of ciprofloxacin-resistant (CIPR) E. coli in 13 types of edible raw vegetables, along with their irrigation water and soil in Shaanxi, China. Of 349 samples collected (157 vegetables, 59 water, and 133 soil), a total of 48 positive samples were detected, with one CIPRE. coli strain isolated from each sample being selected for further analyses. A striking observation was its high prevalence in irrigation water at 44.1 %, markedly exceeding that in vegetables (12.0 %) and soil (4.5 %). The susceptibility of Forty-eight CIPRE. coli isolates was evaluated using the disc diffusion method for 18 different antibiotics, all these isolates were not only resistant to the tested fluoroquinolones antibiotics (levofloxacin, nalidixic acid), but also displayed a multi-drug resistance (MDR) pattern. Twenty-eight (58.3 %) of 48 CIPRE. coli isolates exhibited extended spectrum β-lactamases (ESBLs) (CIPR-ESBLs) producing phenotype. Subsequently, whole-genome sequencing was performed on these 28 isolates. We identified 12 serotypes and STs each, with O101: H9 (35.7 %, 10/28) and ST10 (21.4 %, 6/28) being the most common. Further classification placed these isolates into five phylogenetic groups: A (57.1 %, 16/28), B1 (32.1 %, 9/28), D (3.6 %, 1/28), B2 (3.6 %,1/28), and F (3.6 %,1/28). Notelly, Identical ST types, serotypes and phylogroups were found in certain CIPR-ESBLs-producing E. coli from both vegetables and adjacent irrigation water. Genomic analysis of the 28 CIPR-ESBLs-producing E. coli isolates unveiled 73 resistance genes, associated with 13 amino acid mutations in resistance-determining regions (QRDRs) and resistance to 12 types of antibiotics. Each isolate was confirmed to carry both ESBLs and fluoroquinolone resistance genes, with the Ser83Ala mutation in GyrA (96.4 %, 27/28) being the most prevalent. A detailed analysis of Mobile Genetic Elements (MGEs) revealed that IncFIB and IncFII plasmid subtypes were most prevalent in 60.7 % and 67.9 % of isolates, respectively, with 75 % containing over 10 insertion sequences (IS) each. Furthermore, we observed that certain ESBL and PMQR genes were located on plasmids or in proximity to insertion sequences. In conclusion, our research highlights the widespread presence of CIPRE. coli in irrigation water and thoroughly examines the genetic characteristics of CIPR-ESBLs-producing E. coli strains, underlining the need for ongoing monitoring and management to reduce multidrug-resistant bacteria in vegetables and their environment.

Salmonella is a major foodborne pathogen that poses a substantial risk to food safety and public health. This study aimed to assess the prevalence, antibiotic susceptibility, and genomic features of Salmonella isolates recovered from 600 retail meat samples (300 pork, 150 chicken and 150 beef) from August 2018 to October 2019 in Shaanxi, China. Overall, 40 (6.67 %) of 600 samples were positive to Salmonella, with the highest prevalence in chicken (21.33 %, 32/150), followed in pork (2.67 %, 8/300), while no Salmonella was detected in beef. A total of 10 serotypes and 11 sequence types (STs) were detected in 40 Salmonella isolates, with the most common being ST198 S. Kentucky (n = 15), ST13 S. Agona (n = 6), and ST17 S. Indiana (n = 5). Resistance was most commonly found to tetracycline (82.50 %), followed by to ampicillin (77.50 %), nalidixic acid (70.00 %), kanamycin (57.50 %), ceftriaxone (55.00 %), cefotaxime (52.50 %), cefoperazone (52.50 %), chloramphenicol (50.00 %), levofloxacin (57.50 %), cefotaxime (52.50 %), kanamycin (52.50 %), chloramphenicol (50.00 %), ciprofloxacin (50.00 %), and levofloxacin (50.00 %). All ST198 S. Kentucky isolates showed multi-drug resistance (MDR; ≥3 antimicrobial categories) pattern. Genomic analysis showed 56 distinct antibiotic resistance genes (ARGs) and 6 target gene mutations of quinolone resistance determining regions (QRDRs) in 40 Salmonella isolates, among which, the most prevalent ARG types were related to aminoglycosides and β-lactams resistance, and the most frequent mutation in QRDRs was GyrA (S83F) (47.5 %). The number of ARGs in Salmonella isolates showed a significant positive correlation with the numbers of insert sequences (ISs) and plasmid replicons. Taken together, our findings indicated retail chickens were seriously contaminated, while pork and beef are rarely contaminated by Salmonella. Antibiotic resistance determinants and genetic relationships of the isolates provide crucial data for food safety and public health safeguarding.

This study investigated the prevalence of Salmonella in 210 retail meat samples (105 raw chicken and 105 raw pork) collected from supermarkets and wet markets in 13 areas of Hebei Province, China, from June to October 2018. Whole-genome sequencing was performed on all 125 Salmonella isolates to investigate their genetic relationship. Core genome multilocus sequence typing of 77 representative isolates was used to further elucidate the genetic relatedness among the Salmonella isolated from retail meat. The mean detection rate of Salmonella in all samples was 59.5% (125/210). The prevalence of Salmonella was 53.3% (56/105) in chicken and 65.7% (69/105) in pork. Chicken and pork samples collected in July had the highest detection rate of Salmonella among the sampling months. The isolates were assigned to 19 serotypes, with S. Derby, S. London, and S. Thompson being the most frequent serotypes. Resistance to tetracycline (primarily used for the treatment of bacterial infections) was observed in 89.6% of the isolates, and 84.0% were resistant to doxycycline (also a tetracycline antibiotic) or gemifloxacin (commonly used for clinical treatment of human acute bronchitis). More than 80% of the isolates were multidrug resistant. A total of 21 sequence types were identified. Sequence type 40 (ST-40), the predominant genotype among all isolates, was found only in pork; the sequence types of chicken isolates were more diverse. A total of 58 different antibiotic resistance genes (ARGs) were detected in the 125 isolates. Most types of ARGs were associated with aminoglycoside and β-lactam resistance. Nevertheless, the tetracycline resistance gene tet(A) was the most frequently occurring ARG in all isolates at 78.4%. Multiple isolates of ST-26 contained 20 ARGs. All isolates of ST-40 were divided into two clusters, with at least 160 allelic differences between them. The findings highlight the need to continually monitor ARGs in foodborne Salmonella with particular emphasis on ST-40 and ST-26; the monitoring should include as many retail meat types as possible in the study area.

The aim of this study was to develop homogeneous and stable plasmid DNA reference materials for detecting the mechanisms of resistance to quinolones and fluoroquinolones in foodborne pathogens. The DNA fragments of 11 target genes associated with quinolone and fluoroquinolone resistance were artificially synthesized, inserted into plasmid vectors, and transferred into recipient cells. PCR and sequencing of DNA were performed to assess the genetic stability of the target DNA in recombinant Escherichia coli DH5α cells during subculturing for 15 generations. The limit of detection (LOD) of the target DNA was determined using PCR and real-time qualitative PCR (qPCR). The homogeneity and storage stability of plasmid DNA reference materials were evaluated in terms of plasmid DNA quantity, PCR-measured gene expression, and qPCR threshold cycle. All 11 target DNAs were successfully synthesized and inserted into vectors to obtain recombinant plasmids. No nucleotide mutations were identified in the target DNA being stably inherited and detectable in the corresponding plasmids during subculturing of recombinant strains. When the target DNA was assessed using PCR and qPCR, the LOD was ≤1.77 × 105 and 3.26 × 104 copies/μL, respectively. Further, when the reference materials were stored at 37 °C for 13 days, 4 °C for 90 days, and −20 °C for 300 days, each target DNA was detectable by PCR, and no mutations were found. Although the threshold cycle values of qPCR varied with storage time, they were above the LOD, and no significant differences were found in the quantity of each plasmid DNA at different timepoints. Further, the homogeneity and stability of the materials were highly consistent with the requirements of standard reference materials. To summarize, considering that our plasmid DNA reference materials conformed to standard requirements, they can be used to detect the mechanisms of quinolone and fluoroquinolone resistance in foodborne pathogens.