Recognition of pathogens in food matrixes based on the untargeted in vivo microbial metabolite profiling via a novel SPME/GC × GC-QTOFMS approach.

Foodborne diseases incurred by pathogenic bacteria are one of the major threats in food safety, and thus it is important to develop facile and effective recognition methodology of pathogens in food. Herein, a new automatic approach for detection of in vivo volatile metabolites emitted from foodborne pathogens was proposed by coupling solid phase microextraction (SPME) technique with a comprehensive two-dimensional gas chromatography quadrupole time-of-flight mass spectrometry (GC × GC-QTOFMS). A novel polymer composite based SPME probe which possessed high-coverage of microbial metabolites was utilized in this contribution to realize the sensitive extraction of untargeted metabolites. As a result, a total of 126 in vivo metabolites generated by the investigated pathogens were detected and identified, with 33, 29, 25, 21 and 18 volatile metabolites belonging to Shigella sonnei, Escherichia coli, Salmonella typhimurium, Vibrio parahaemolyticus and Staphylococcus aureus, respectively. Multivariate statistical analyses were applied for further analysis of metabolic data and separation of responsive metabolic features among different microbial systems were found, which were also successfully verified in foodstuffs contaminated by microorganisms. The growth trend of the potential volatile markers of each pathogen in food samples kept consistent with that of the pure strain incubated in medium during the whole incubation time. This study promotes the application of SPME technology in microbial volatile metabolomics and contributes to the development of new approaches for foodborne pathogens recognition.

Synthesis of carboxymethylated ß-glucan from naked barley bran and its antibacterial activity and mechanism against Staphylococcus aureus.

This study aimed to elucidate the antimicrobial efficacy of carboxymethylated ß-glucan (CMG) against Staphylococcus aureus and the possible underlying mechanisms. ß-Glucan (BG) was extracted from naked barley bran, then ultrafiltrated to obtain four fractions (BG-1, BG-2, BG-3, BG-4). Five types of CMG were prepared using an amidation reaction and characterized by FTIR spectroscopy. The underlying mechanisms were studied using changes in the transmembrane electrical potential, pH gradient, and intracellular ATP, confocal laser scanning microscopy, and electron microscopy. The data showed that CMG-2 had the highest antibacterial activity. CMG-2 induced membrane permeabilization, morphological changes, and structural disruption in S. aureus cells, with the formation of blebs and microspheres, the disruption of cell walls and membranes, and cell lysis. Our findings provide insight into the antimicrobial mechanism of CMG against S. aureus, which will provide a foundation for developing CMG as a novel antibacterial agent in the food and biomedical fields.

Chemical characteristics and antithrombotic effect of chondroitin sulfates from sturgeon skull and sturgeon backbone.

Chondroitin sulfates (CSs) were extracted from sturgeon skull and backbone, and their chemical composition, anticoagulant, anti-platelet and thrombolysis activities were evaluated. The average molecular weights of CS from sturgeon skull and backbone were 38.5kDa and 49.2kDa, respectively. Disaccharide analysis indicated that the sturgeon backbone CS was primarily composed of disaccharide monosulfated in position four of the GalNAc (37.8%) and disaccharide monosulfated in position six of the GalNAc (59.6%) while sturgeon skull CS was primarily composed of nonsulfated disaccharide (74.2%). Sturgeon backbone CS showed stronger antithrombotic effect than sturgeon skull CS. Sturgeon backbone CS could significantly prolong activated partial thromboplastin time (APTT) and thrombin time (TT), inhibited ADP-induced platelet aggregation and dissolved platelet plasma clots in vitro. The results suggested that sturgeon backbone CS can be explored as a functional food with antithrombotic function.

Paraplantaricin L-ZB1, a novel bacteriocin and its application as a biopreservative agent on quality and shelf life of rainbow trout fillets stored at 4 °c.

Paraplantaricin L-ZB1 was produced by Lactobacillus paraplantarum L-ZB1, which was isolated from the traditional China fermented sausage. In this work, paraplantaricin L-ZB1 was used to maintain quality of rainbow trout fillets at 4 °C. Rainbow trout fillets were left untreated (CK) or treated with 200 activity units (AU)/ml paraplantaricin L-ZB1 (P1), 400 AU/ml paraplantaricin L-ZB1 (P2) or 200 AU/ml Nisin (N). The treated samples were stored at 4 °C for up to 10 days, and the quality changes were determined by microbiological (total viable count [TVC], Enterobacteriaceae, Pseudomonas, spore-forming bacteria), sensory, chemical (pH, total volatile basic nitrogen [TVB-N]) and biochemical (biogenic amines, K value) methods. Results show that paraplantaricin L-ZB1 could inhibit the growth of microflora, especially Enterobacteriaceae, Pseudomonas and spore-forming bacteria during sample storage. Meanwhile, the increases of pH, TVB-N, K value and biogenic amine levels were significantly delayed in paraplantaricin L-ZB1-treated samples compared to the control group. Paraplantaricin L-ZB1 of 400 AU/ml extended the rainbow trout fillets' shelf life to 4-6 days, and the sample showed good sensory characteristics. Therefore, paraplantaricin L-ZB1 could be used as a suitable biological preservative for chilled rainbow trout fillets.

Biogenic amines formation, nucleotide degradation and TVB-N accumulation of vacuum-packed minced sturgeon (Acipenser schrencki) stored at 4 °C and their relation to microbiological attributes.

BACKGROUND: Sturgeon (Acipenser schrencki) were filleted, minced, vacuum-packed and stored at 4 °C. Minced sturgeon muscles were analysed for 10 days to determine biochemical [eight biogenic amines, ATP and breakdown products, K value, total volatile basic nitrogen (TVB-N) and pH], microbiological (total viable counts, Enterobacteriaceae, Psychrotrophs, Pseudomonas, Aeromonas, Vibrio, Lactobacillus and Brochothrix thermosphacta) changes and their relationships. RESULTS: The shelf-life of sturgeon was found to be 3-4 days according to microbiological assessment. At the end of the storage, Psychrotrophs, Enterobacteriaceae and Aeromonas became the dominant bacteria, reaching 7.85, 7.98 and 8.02 log cfu g(-1), respectively. Biogenic amines, hypoxanthine and the K value increased significantly (P < 0.05) during the storage. Putrescine and cadaverine was the dominant biogenic amines while tryptamine was not detected during the storage. In addition, putrescine, cadaverine and hypoxanthine were highly related to storage time and total viable counts (r(2), 0.876-0.962), indicating that they can be good biomarkers for quality assessment of vacuum-packed minced sturgeon stored at 4 °C. CONCLUSION: In this study, key biochemical and microbiological attributes were established to monitor the spoilage and shelf life of vacuum-packed sturgeon. These results could help to establish better cold storage management of minced sturgeon products under commercial conditions.