Metagenomic profiles of the antimicrobial resistance in traditional Chinese fermented meat products: Core resistome and co-occurrence patterns.

Antimicrobial resistance (AMR) poses a significant challenge to global health, and the presence of antibiotic resistance genes (ARGs) in food poses a potential threat to public health. Traditional Chinese fermented meat products (FMPs) are highly favored because of their unique flavors and cultural value. However, microbial safety and the potential distribution and composition of AMR in these products remain unclear. In this study, a comprehensive analysis of bacterial composition and antibiotic-resistant populations in 216 samples of traditional fermented meat products from different regions of China was conducted using a metagenomic approach. Staphylococcus was the most abundant genus in the samples, accounting for an average abundance of 29.9 %, followed by Tetragenococcus (17.1 %), and Latilactobacillus (3.6 %). A core resistome of FMP samples was constructed for the first time using co-occurrence network analysis, which revealed the distribution and interrelationships of ARGs and bio/metal-resistant genes (BMRGs). Random forest analysis identified the lincosamide nucleotidyltransferase lnuA and the multidrug and toxic compound extrusion (MATE) transporter abeM as potential indicators for assessing the overall abundance of the core resistome. Additionally, Staphylococcus, Acinetobacter, and Pseudomonas were identified as hosts constituting the core resistome. Despite their low abundance, the latter two still serve as major reservoirs of antibiotic resistance genes. Notably, Lactococcus cremoris was identified as the key host for tetracycline resistance genes in the samples, highlighting the need for enhanced resistance monitoring in lactic acid bacteria. Based on our findings, in the microbial safety assessment of fermented meat products, beyond common foodborne pathogens, attention should be focused on detecting and controlling coagulase-negative Staphylococcus, Acinetobacter, and Pseudomonas, and addressing bacterial resistance. The quantitative detection of lnuA and abeM could provide a convenient and rapid method for assessing the overall abundance of the core resistome. Our findings have important implications for the control of bacterial resistance and prevention of pathogenic bacteria in fermented meat products.

Metagenomics reveals differences in the composition of bacterial antimicrobial resistance and antibiotic resistance genes in pasteurized yogurt and probiotic bacteria yogurt from China.

Antimicrobial resistance has become a global public health concern, and antibiotic resistance genes (ARG) in food are a research focus. In China, probiotics and pasteurized yogurts are the 2 main types of commercially available yogurt, but the distribution and differences of antibiotic-resistant bacteria and gene types in these products are not well known. This study used a shotgun metagenomic approach to analyze 22 different types of yogurt collected from 9 main yogurt-producing areas in China; each type of yogurt included 8 different batches of samples. The abundance and diversity of bacteria identified in probiotic yogurt were significantly higher than those in pasteurized yogurt, with Acetobacter, Raoultella, and Burkholderia identified as unique and highly abundant genera in probiotic yogurt. Similarly, the abundance of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp. was higher than that in pasteurized yogurt. A total of 1,149 ARG subtypes belonging to 16 ARG types were identified, with the highest abundance of rifampicin, multidrug efflux pumps, and quinolone resistance genes detected. Network analysis revealed significant nonrandom co-occurrence relationships between different types and subtypes of ARG in yogurt samples. A total of 44 ARG subtypes in pasteurized yogurt were potentially hosted by 36 bacterial genera, and in probiotic yogurt, 63 ARG were expected to be hosted by 86 bacterial species from 37 genera. These findings indicate potential safety issues in fermented dairy products and emphasize the need for a more hygienic environment when processing probiotic yogurt.

Single-molecule real-time sequencing reveals differences in bacterial diversity in raw milk in different regions and seasons in China.

The quality of raw milk is a key factor influencing the whole dairy processing chain. The richness and diversity of bacteria in raw milk affect its quality and safety. However, traditional microbial detection methods mainly depend on the known microbe culture and are often time consuming. Thus, the development of efficient ways for supervising any possible microbiological contamination is desiderated. In the current work, single-molecule real-time (SMRT) sequencing, developed by Pacific Biosciences (PacBio), was applied to acquire long reads and applied for discrimination of bacteria at species level. Forty samples of raw milk obtained from Beijing, Hebei, Inner Mongolia, Shanghai, and Guangdong in China during summer, autumn, and winter were investigated. Among 35 bacteria species identified in these samples, Acinetobacter albensis, Pseudomonas gessardii, Pseudomonas weihenstephanensis, and Rahnella inusitata were the bacteria with the highest relative abundance in the overall sample, whereas the bacteria with the highest relative abundance in raw milk samples of different origins and seasons are different. Significant differences in bacterial richness and bacterial community diversity in raw milk grouped according to different production areas and different sampling seasons were confirmed by Welch's t-test. Interestingly, the transport distance and transport time positively correlated with the relative abundance of Pseudomonas weihenstephanensis, suggesting that the content of this bacteria was expected to be a standard for evaluating the freshness of raw milk. Pathogens Bacillus cereus and Klebsiella pneumoniae were detected in most samples, indicating that the raw milk was at risk of contamination by pathogenic bacteria. Moreover, the findings of this study provide important evidence for quality and safety monitoring and biological control of raw milk.

Application of metabolomics analysis to aid in understanding the pathogenicity of different lineages and different serotypes of Listeria monocytogenes.

Listeria monocytogenes is a foodborne pathogen with high mortality in young children, elderly persons, pregnant women, and immune-compromised individuals. Most human listeriosis cases are associated with four serotypes (1/2a, 1/2b, 1/2c and 4b) within lineages I and II. The intracellular metabolic changes in L. monocytogenes from different lineages and serotypes remain unclear. Here, a non-targeted metabolomics strategy, based on ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) coupled with chemometrics was performed to analyze the metabolic fluctuations in the lineages and serotypes of L. monocytogenes. In the aggregate, 93 L. monocytogenes isolates (two lineages and four serotypes) were investigated, and a total of 48 differential biomarker metabolites were identified. Twenty differential metabolites were observed at the lineage level, while the others were found at the serotype level. At the lineage level, significantly lower contents (fold change [FC] = 3.60) of γ-aminobutyric acid (GABA) was found in lineage II strains, as compared to lineage I strains. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways enrichment analysis showed that different substances between lineages I and II participated in multiple metabolic pathways, among which pyrimidine metabolism, purine metabolism, and aminoacyl-tRNA biosynthesis were significantly enriched. The major significantly upregulated (P < 0.05) metabolites, including uridine 5'-monophosphate, uridine, guanosine monophosphate, and guanosine, in lineage II strains affect the synthesis of DNA and RNA. At the serotype level, the upregulation of urea precursor substances produced by the purine metabolism pathway in serotype 4b strains may be one reason for the increased virulence of serotype 4b strains, compared to other serotype strains. In addition, the upregulation of pyridoxine in the vitamin B6 pathway in 4b strains was also observed. The differential metabolic pathway of 1/2a and 1/2c strains is nucleotide metabolism, which could lead to differences in the biosynthesis of DNA and RNA of the strain and further affect the growth and proliferation of strains.

Insight into Bacillus cereus Associated with Infant Foods in Beijing.

This study was undertaken to investigate the prevalence, antimicrobial resistance, and virulence gene profiles of Bacillus cereus in different brands of infant formula in Beijing supermarkets. Eighty-eight Bacillus cereus isolates were recovered in sixty-eight infant formulas of five domestic brands and fourteen imported brands. The prevalence rate in domestic and imported samples were 70.6% and 52.9%, respectively. Lower mean prevalence level was found in domestic samples (1.17 MPN/g) compared with the imported samples (3.52 MPN/g). Twenty-four virulence gene profiles were found, and most strains carried at least one virulence gene. The prevalence of nheA, nheB, nheC, cytK, bceT, and entFM in domestic and imported brand samples was similar. The occurrence of enterotoxin genes hblA, hblC, and hblD in domestic samples were 22.2%, 27.8%, and 22.2%, respectively, which was significantly higher than imported samples. Antimicrobial drugs-susceptibility analysis showed that all isolates were susceptible to gentamincin, amikacin, and ciprofloxacin; 38%, 7%, and 2.3% were resistant to rifampin, tetracycline, and chloramphenicol, respectively; and only one isolate was resistant to trimethoprim-sulfamethoxazole. Moreover, the cell numbers of Bacillus cereus in prepared infant formula increased rapidly at room temperature. Thus, monitoring guidelines are needed for accepted levels of Bacillus cereus in infant formula.

Enhanced response of sensor on serotonin using nickel-reduced graphene oxide by atomic layer deposition.

Atomic layer deposition (ALD) is a promising method for preparing nanomaterials. The thickness and uniformity of nanomaterials can be precisely controlled. Hence, the uniform Ni nanoparticles (Ni NPs) deposited on reduced graphene oxide (rGO) by ALD and got the optimal combination interface. The morphology, structure, and electrochemical behavior of Ni NPs-rGO nanocomposite are investigated. By experiment results, the Ni NPs could occupy some active surface of rGO, resulting in high conductivity and large specific surface area of Ni NPs-rGO nanocomposite. The Ni NPs-rGO nanocomposite exhibits high electrocatalytic activity for serotonin and speeds up the electron transfer between the surface of the electrode and the solution. Therefore, the sensor is prepared by Ni NPs-rGO nanocomposite modified glassy carbon electrode (GCE) and used to sensitive detection of serotonin. By differential pulse voltammetric, the Ni NPs-rGO/GCE enhanced the current responses and showed a wide linear range of 0.02-2 µM with a low detection of 0.01 µM for serotonin (S/N = 3). The Ni NPs-rGO/GCE exhibited good stability, selectivity, and anti-interference ability that can be used for real sample detection. According to these results, the Ni NPs-rGO nanocompositeis successfully prepared by ALD. The properties of Ni NPs-rGO nanocomposite make it an attractive material for potential applications in sensors and catalysis.

Response of Formed-Biofilm of Enterobacter cloacae, Klebsiella oxytoca, and Citrobacter freundii to Chlorite-Based Disinfectants.

Bacterial biofilms formed on equipment surfaces are potential sources of cross-contamination and can be responsible for the spread of bacteria involved in food spoilage, such as some Enterobacteriaceae family members. In this study, the effect of chlorite-based disinfectants, including sodium hypochlorite (SH), chlorine dioxide (CD), strongly acidic electrolyzed water (StAEW), and neutral electrolyzed water (NEW), on inactivation of mono-biofilms of Enterobacter cloacae, Klebsiella oxytoca, and Citrobacter freundii was evaluated separately. All the strains were enumerated by the viable plate-count method after disinfection for 30 min. A comparison of the surviving cells after disinfection indicated that E. cloacae biofilms were more resistant to disinfectants than the biofilms of the other two strains, and treatment with all the disinfectants improved sanitizing. SH (200 mg/L) was the most effective in the reduction of cell number in the biofilms of all strains. Considering the safety of use and environmental protection, electrolyzed oxidizing water, especially StAEW, was a good suggestion for the inactivation of cells in K. oxytoca or C. freundii biofilms. These results suggest that the cells in biofilm of E. cloacae, K. oxytoca, and C. freundii were highly sensitive to chlorite-based disinfectants and provide insights into the efficacy of disinfectants in killing bacteria. PRACTICAL APPLICATION: The Enterobacteriaceae biofilms formed on equipment surfaces, which can cause cross-contamination and food spoilage, are greatly challenging bacterial contaminants of food products. Electrolyzed oxidizing water is a novel, environmentally friendly disinfectant that can effectively treat Enterobacteriaceae biofilms. The results of this study may be used to design effective measures to disinfect biofilms on equipment contact surfaces.

Preliminary Transcriptome Analysis of Mature Biofilm and Planktonic Cells of Salmonella Enteritidis Exposure to Acid Stress.

Salmonella has emerged as a well-recognized food-borne pathogen, with many strains able to form biofilms and thus cause cross-contamination in food processing environments where acid-based disinfectants are widely encountered. In the present study, RNA sequencing was employed to establish complete transcriptome profiles of Salmonella Enteritidis in the forms of planktonic and biofilm-associated cells cultured in Tryptic Soytone Broth (TSB) and acidic TSB (aTSB). The gene expression patterns of S. Enteritidis significantly differed between biofilm-associated and planktonic cells cultivated under the same conditions. The assembled transcriptome of S. Enteritidis in this study contained 5,442 assembled transcripts, including 3,877 differentially expressed genes (DEGs) identified in biofilm and planktonic cells. These DEGs were enriched in terms such as regulation of biological process, metabolic process, macromolecular complex, binding and transferase activity, which may play crucial roles in the biofilm formation of S. Enteritidis cultivated in aTSB. Three significant pathways were observed to be enriched under acidic conditions: bacterial chemotaxis, porphyrin-chlorophyll metabolism and sulfur metabolism. In addition, 15 differentially expressed novel non-coding small RNAs (sRNAs) were identified, and only one was found to be up-regulated in mature biofilms. This preliminary study of the S. Enteritidis transcriptome serves as a basis for future investigations examining the complex network systems that regulate Salmonella biofilm in acidic environments, which provide information on biofilm formation and acid stress interaction that may facilitate the development of novel disinfection procedures in the food processing industry.

Visual and light scattering spectrometric method for the detection of melamine using uracil 5'-triphosphate sodium modified gold nanoparticles.

A highly selective method was presented for colorimetric determination of melamine using uracil 5'-triphosphate sodium modified gold nanoparticles (UTP-Au NPs) in this paper. Specific hydrogen-bonding interaction between uracil base (U) and melamine resulted in the aggregation of AuNPs, displaying variations of localized surface plasmon resonance (LSPR) features such as color change from red to blue and enhanced localized surface plasmon resonance light scattering (LSPR-LS) signals. Accordingly, the concentration of melamine could be quantified based on naked eye or a spectrometric method. This method was simple, inexpensive, environmental friendly and highly selective, which has been successfully used for the detection of melamine in pretreated liquid milk products with high recoveries.

Ratiometric fluorescence recognition for pyrophosphate on the basis of terpyridine derivative.

Diphosphate (pyrophosphate, PPi) is vital for organisms, and therefore its detection is of special importance. In this paper, one cadmium complex of terpyridine (tpy) derivative, 4'-(aminomethylphenyl)-2,2':6',2"-terpyridine (aptpy), has been reported for the ratiometric fluorescence recognition of PPi. When added with cadmium, the emission of aptpy at 358 nm was greatly enhanced and red shifted to 397 nm due to the complexation-induced ICT process, which then blue shifted to 349 nm upon the further addition of PPi. Based on the different response of dual fluorescence emissions at 349 and 397 nm, a ratiometric fluorescence method could be successfully established for the fluorescence recognition of PPi. With that, PPi could be successfully discriminated from other structurally similar anions, including nucleotide triphosphates.

[Study on panchromatic band broadening of new high-resolution satellite sensor].

For developing a remote sensor, the selection of operating waveband is one of the most important factors for detecting and identifying target. In the present paper, the changes of atmospheric effects and imagery quality are simulated due to the increase in the response wave range of optical remote sensor from 0.50-0.85 mm to 0.45-0.90 mm by using MODTRAN4. The experimental results show that there is a slight increase of the adverse factors, including atmospheric transmittance, path radiance, and adjacency effect, after the working waveband has been widened. The disadvantages compared with the improvement in incident radiance, target-background contrast and image quality are negligible. In summary, the scheme of 0.45-0.90 mm is superior to 0.50-0.85 mm and it has been more widely used in the on-orbit operation high-resolution satellite sensor.