Foodborne Pathogenic Bacteria: Prevalence and Control-Volume I.

From the farm to the dining table, foodborne pathogenic bacteria can contaminate food at any stage of the food production, processing, delivery, preparation, and consumption chain, posing a critical threat to the safety of food systems worldwide [...].

Prevalence, antimicrobial resistance, and genetic characteristics of Staphylococcus aureus isolates in frozen flour and rice products in Shanghai, China.

Staphylococcus aureus is widely recognized as a highly hazardous pathogen that poses significant threats to food safety and public health. This study aimed to assess the prevalence, antimicrobial resistance, and genetic characteristics of S. aureus isolates recovered from 288 frozen flour and rice product samples in Shanghai, China, between September 2019 and May 2020. A total of 81 S. aureus isolates were obtained, representing 25 sequence types (STs), with ST7 being the most prevalent (17.28%, n = 14). The majority of S. aureus isolates (85.19%, n = 69) carried at least one enterotoxin gene, with the seg gene being the most frequently detected (51.85%, n = 42). Additionally, 12 isolates (14.81%) were identified as methicillin-resistant S. aureus (MRSA) through mecA gene detection. Notably, this study reported the presence of an ST398 MRSA isolate in frozen flour and rice products for the first time. All MRSA isolates displayed multidrug resistance, with the highest resistance observed against cefoxitin (100.00%), followed by penicillin (91.67%) and erythromycin (66.67%). Genomic analysis of the 12 MRSA isolates revealed the presence of twenty distinct acquired antimicrobial resistance genes (ARGs), eight chromosomal point mutations, and twenty-four unique virulence genes. Comparative genome analysis indicated close genetic relationships between these MRSA isolates and previously reported MRSA isolates from clinical infections, highlighting the potential transmission of MRSA through the food chain and its implications for public health. Significantly, the identification of three plasmids harboring ARGs, insertion sequences (ISs), the origin of transfer site (oriT), and the relaxase gene suggested the potential for horizontal transfer of ARGs via conjugative plasmids in S. aureus. In conclusion, this study revealed significant contamination of retail frozen flour and rice products with S. aureus, and provided essential data for ensuring food safety and protecting public health.

Correlation Analysis between GlpQ-Regulated Degradation of Wall Teichoic Acid and Biofilm Formation Triggered by Lactobionic Acid in Staphylococcus aureus.

Staphylococcus aureus biofilms are a serious problem in the food industry. Wall teichoic acid (WTA) is crucial in S. aureus biofilm formation. Overexpression of the WTA-hydrolyzing enzyme glycerophosphoryl diester phosphodiesterase (GlpQ), induced by lactobionic acid (LBA), may be related to biofilm formation. We investigated the relationship between the regulation on GlpQ degradation of WTA by LBA and S. aureus biofilm formation. LBA minimum inhibitory concentration for S. aureus was 12.5 mg/mL. Crystal violet staining revealed the LBA-mediated inhibition of S. aureus adhesion and biofilm formation. RT-qPCR revealed the repressed expression of adhesion-related genes by LBA. Scanning electron microscopy revealed the obvious disruption of S. aureus surface structure, confirming the repression of S. aureus adhesion and biofilm formation by LBA. Native-PAGE results suggested that the WTA content of S. aureus was reduced under the inhibition of LBA. Additionally, LBA induced the overexpression of glpQ. Combined with our previous work, these results suggest that glpQ is induced in S. aureus to function in WTA degradation with the addition of LBA, resulting in decreased WTA content and subsequent reduction of adhesion and biofilm formation. The findings provide new insight into the degradation mechanism of S. aureus WTA and indicate the potential of LBA as an anti-biofilm agent.

The non-covalent interactions between whey protein and various food functional ingredients.

In daily diet, Whey protein (WP) is often coexisted with various Food functional ingredients (FFI) such as proteins, polyphenols, polysaccharides and vitamins, which inevitably affect or interact with each other. Generally speaking, they may be interact by two different mechanisms: non-covalent and covalent interactions, of which the former is more common. We reviewed the non-covalent interactions between WP and various FFI, explained the effect of each WP-FFI interaction, and provided possible applications of WP-FFI complex in the food industry. The biological activity, physical and chemical stability of FFI, and the structure and functionalities of WP were enhanced through the non-covalent interactions. The development of non-covalent interactions between WP and FFI provides opportunities for the design of new ingredients and biopolymer complex, which can be applied in different fields. Future research will further focus on the influence of external or environmental factors in the food system and processing methods on interactions.

Whey protein and xylitol complex alleviate type 2 diabetes in C57BL/6 mice by regulating the intestinal microbiota.

Type 2 diabetes (T2D) is a metabolic disorder that has become a major threat to public health. Epidemiological and experimental studies have suggested that whey protein isolate (WPI) and xylitol (XY) play an important role on T2D. This manuscript hypothesizes the supplementation of whey protein and xylitol complex (WXY) has the hypoglycemic and hyperlipidemia effect of T2D mice induced by the conjoint action of a high-fat diet and streptozotocin (STZ) by modulating of intestinal microbiota. The mice with diabetes displayed higher levels of fasting blood glucose (FBG), insulin, glycosylated hemoglobin, total triglycerides, total cholesterol, aspartate aminotransferase, alanine aminotransferase and other serum parameters than the normal mice. Treatment with WXY for 6 weeks significantly modulated the levels of FBG and insulin, improved insulin sensitivity, pancreas impairment and liver function in T2D mice, and the effect was better than that observed with WPI and XY groups. Moreover, supplementation with WXY significantly changed the diversity and composition of the intestinal microbiota in T2D mice and restored the intestinal bacteria associated with T2D (Firmicutes, Bacteroidetes, and Lactobacillus). This may be a potential mechanism for alleviating T2D symptoms. Spearman correlation analysis showed that the relative abundances of specific genera (Turicibacter, Lachnospiraceae_NK4A136_group, Lactobacillus, Candidatus_Saccharimonas, Faecalibaculum and Coriobacteriaceae_UCG-002) were correlated with the levels of blood glucose and serum parameters. Therefore, WXY may be considered a promising dietary supplement for T2D treatment in the future.

Characterization and comparison of lipids in bovine colostrum and mature milk based on UHPLC-QTOF-MS lipidomics.

Lipids in bovine milk have several biological activities, with implications for human health and the physical functionality of foods. However, alterations in the lipid profile of bovine milk during lactation are not well-studied. This study aimed to identify differences in lipids between bovine colostrum and mature milk, using a lipidomics approach. Using an advanced mass spectrometry-based quantitative lipidomics approach, 335 lipids assigned to 13 subclasses were characterized in bovine colostrum (BC) and mature milk (BM). In total, 63 significantly differential lipids (SDLs) were identified. Among the 63 SDLs, the levels of 21 lipids were significantly lower in BM than in BC, including 5 glycerophosphatidylethanolamines (PEs), 1 glycerophosphatidylglycerol (PG), and 15 triacylglycerols (TGs). The levels of the remaining 42 lipids increased in BM, including 1 cardiolipin (CL), 9 diacylglycerols (DGs), 9 dihexosylceramides (Hex2Cers), 3 hexosylceramides (HexCers), 3 glycerophosphatidic acids (PAs), 2 glycerophosphatidylcholines (PCs), 12 PEs, and 3 TGs. Furthermore, the correlations and related metabolic pathways of these 63 SDLs were analyzed to explore the mechanisms that alter bovine milk lipids during lactation. The seven most relevant pathways identified herein, ranked in accordance with their degree of influence on lactation, were glycerophospholipid metabolism, sphingolipid metabolism, glycerolipid metabolism, glycosylphosphatidylinositol-anchor biosynthesis, linoleic acid metabolism, alpha-linolenic acid metabolism, and arachidonic acid metabolism. Our results provide essential insights into mechanisms underlying alterations in bovine milk lipids during different lactation periods, along with practical information of specific nutrition and quality assessments for the dairy industry.

Interaction of xylitol with whey proteins: Multi-spectroscopic techniques and docking studies.

This study aimed to examine the interaction mechanism between xylitol (XY) and whey protein isolate (WPI) using multispectral techniques and molecular docking. Additionally, we investigated the effect of XY on WPI functionality using the method of fluorescent probe, high-speed dispersion and differential scanning calorimetry. The fluorescence quenching results such as quenching constants, binding constants and thermodynamic parameters showed strong susceptibility to interacting of WPI and its fractions to XY and the sequence was: α-lactalbumin (α-La) > bovine serum albumin (BSA) > ß-lactoglobulin (ß-Lg). Docking results revealed that XY was bound to the residues of aromatic cluster II in α-La, the hydrophobic cavity of ß-Lg and the subdomain IIA of BSA through hydrogen bonding and van der Waals forces, resulting in conformational changes in secondary structures of proteins, which converted α-helix to ß-turn and random coils. Further, XY increased thermal stability and emulsifying properties and reduced surface hydrophobicity and zeta-potential of WPI.

Comparative metabolomics analysis of donkey colostrum and mature milk using ultra-high-performance liquid tandem chromatography quadrupole time-of-flight mass spectrometry.

Donkey milk has been widely shown to be an ideal substitute for human milk because of its similar composition. However, alterations to the composition of donkey milk during lactation have not been well studied. In this study, untargeted metabolomics with ultra-high-performance liquid tandem chromatography quadrupole time-of-flight mass spectrometry were used to analyze and compare the metabolites in donkey colostrum (DC) and mature milk (DMM). Two hundred seventy metabolites were characterized in both DC and DMM. Fifty-two of the metabolites in the DC were significantly different from those in the DMM; 8 were downregulated and 44 were upregulated. This demonstrated that the composition of the donkey milk changed with lactation. Additionally, the interactions and metabolic pathways were further analyzed to explore the mechanisms that altered the milk during lactation. Our results provide comprehensive insights into the alterations in donkey milk during lactation. The results will aid in future investigations into the nutrition of donkey milk and provide practical information for the dairy industry.

Quantitative lipidomics reveals alterations in donkey milk lipids according to lactation.

The composition of donkey milk is similar to that of human milk. However, the lipid content in donkey milk is lower than that in human milk. Thus far, the lipid composition of donkey milk during lactation has not been well-studied. Through mass spectroscopy-based quantitative lipidomics, we analyzed lipids in donkey colostrum (DC) and mature milk (DM). Thirteen subclasses of 335 lipids were identified in both DC and DM; 60 lipids - 17 upregulated and 43 downregulated - were differentially regulated between DM and DC (Variable Importance in Projection >1, P < 0.05), demonstrating that lipid composition changed with lactation. These different lipids were involved in 19 metabolic pathways, of which glycerophospholipid, linoleic acid, alpha-linolenic acid, glycosylphosphatidylinositol-anchor, glycerolipid, and arachidonic acid metabolism were the most relevant. Our results provide insights into quantitative alterations in donkey milk lipids during lactation, development of donkey milk products, and screening of potential biomarkers.

Label-Free Quantitative Proteomics Reveals the Multitargeted Antibacterial Mechanisms of Lactobionic Acid against Methicillin-Resistant Staphylococcus aureus (MRSA) using SWATH-MS Technology.

The objective of the present study was to reveal the antibacterial mechanism of lactobionic acid (LBA) against methicillin-resistant Staphylococcus aureus (MRSA) using quantitative proteomics by sequential window acquisition of all theoretical mass spectra (SWATH-MS) to analyze 100 differentially expressed proteins after LBA treatment. Furthermore, multiple experiments were conducted to validate the results of the proteomic analysis including reactive oxygen species (ROS), virulence-associated gene expression, and the relative quantification of target proteins and genes by parallel reaction monitoring and quantitative real-time PCR. Combining the ultrastructure observations, proteomic analysis, and our previous research, the mode of LBA action against MRSA was speculated as cell wall damage and loss of membrane integrity; inhibition of DNA repair and protein synthesis; inhibition of virulence factors and biofilm production; induction of oxidative stress; and inhibition of metabolic pathways. These results suggest potential applications for LBA in food safety and pharmaceuticals, considering its multitarget effects against MRSA.

Quantitative N-glycoproteomics of milk fat globule membrane in human colostrum and mature milk reveals changes in protein glycosylation during lactation.

Milk fat globule membrane (MFGM) proteins have recently gained increasing attention, due to their significant biological function. However, the glycosylation of proteins in human MFGM during lactation has not been studied in detail. In this study, through mass spectroscopy-based N-glycoproteomics, we analyzed protein glycosylation of human MFGM. A total of 912 N-glycosylation sites on 506 N-glycoproteins were identified in human colostrum and mature milk MFGM. Among them, 220 N-glycoproteins with 304 N-glycosylation sites were differentially expressed in colostrum and mature milk MFGM. Gene Ontology (GO) analysis revealed various biological processes, cellular components, and molecular functions of the differentially expressed N-glycoproteins. Specifically, these glycoproteins were involved in biological processes such as single-organism processes, biological regulation, regulation of biological processes, response to stimulus and localization; were cellular components in organelles, membranes, and the extracellular region; and had different molecular functions such as protein binding, receptor activity, and hydrolase activity. KEGG pathway analysis suggested that the majority of the differentially expressed N-glycoproteins were associated with phagosome, cell adhesion molecule and some disease-related pathways. Our results provide an in-depth understanding of the quantitative changes in N-glycosylation of proteins in human colostrum and mature MFGM, and extend our knowledge of the N-glycoproteome and of the distribution of N-glycosylation sites in human MFGM during lactation, providing insight into the biological functions of the highlighted glycoproteins.