Pseudomonas weihenstephanensis through the iron metabolism pathway promotes in situ spoilage capacity of prepared beef steaks during cold storage.

In this study, we evaluated the histomorphology, reactive oxygen species (ROS), protein degradation, and iron metabolism characteristics and differential expression analysis of genes for siderophores synthesis and protease secretion in prepared beef steaks inoculated alone or co-inoculated with P. weihenstephanensis, B. thermotrichothrix and M. caseolyticus at 4 °C for 12 days. The results showed that the P. weihenstephanensis was the key bacteria that degraded protein in the process of prepared beef steaks spoilage, which led to protein oxidation by promoting ferritin degradation to release free iron and inducing ROS accumulation. The highest expression of FpvA and AprE was detected in the P. weihenstephanensis group by comparing qRT-PCR of the different inoculation groups. Both qRT-PCR and Western blot revealed that ferritin heavy polypeptide and ferritin light chain polypeptide gene and protein expressions were significantly higher in the P. weihenstephanensis inoculation group compared to the other inoculation groups. Results suggested that FpvA and AprE might play roles in meat spoilage and were potential positional, physiological and functional candidate genes for improving the quality traits of prepared beef steaks. This work may provide insights on controlling food quality and safety by intervening in spoilage pathways targeting iron carrier biosynthesis or protease secretion genes.

Revealing the spoilage characteristics of refrigerated prepared beef steak by advanced bioinformatics tools.

BACKGROUND: Although microorganisms are the main cause of spoilage in prepared beef steaks, very few deep spoilage mechanisms have been reported so far. Aiming to unravel the mechanisms during 12 days of storage at 4 °C affecting the quality of prepared beef steak, the present study investigated the changes in microbial dynamic community using a combined high-throughput sequencing combined and bioinformatics. In addition, gas chromatography-mass spectrometry combined with multivariate statistical analysis was utilized to identify marker candidates for prepared steaks. Furthermore, cloud platform analysis was applied to determine prepared beef steak spoilage, including the relationship between microbiological and physicochemical indicators and volatile compounds. RESULTS: The results showed that the dominant groups of Pseudomonas, Brochothrix thermosphacta, Lactobacillus and Lactococcus caused the spoilage of prepared beef steak, which are strongly associated with significant changes in physicochemical properties and volatile organic compounds (furan-2-pentyl-, pentanal, 1-octanol, 1-nonanol and dimethyl sulfide). Metabolic pathways were proposed, among which lipid metabolism and amino acid metabolism were most abundant. CONCLUSION: The present study is helpful with respect to further understanding the relationship between spoilage microorganisms and the quality of prepared beef steak, and provides a reference for investigating the spoilage mechanism of dominant spoilage bacteria and how to extend the shelf life of meat products. © 2024 Society of Chemical Industry.

"Potential Scalpel": A Bioassisted Ultrafast Staining Lateral Flow Immunoassay from De Novo to Results.

It is of great importance to overcome potential incompatibility problems between dyestuffs and antibodies (mAbs) for extensive commercial application of a dyestuff-chemistry-based ultrafast colorimetric lateral flow immunoassay (cLFIA). Herein, inspired by traditional staining technologies, a basic dyestuff gallocyanine (GC)-assisted biogenic "potential scalpel"-based cLFIA (GC-ABPS-based cLFIA) by employing clenbuterol (CLE) as proof-of-concept was proposed to solve a high degree of incompatibility between the same potential dyestuffs and mAbs. Goat antimouse immunoglobulin (Ab2) could serve as the "potential scalpel" to form the positive potential value biomolecular network self-assemblers (BNSA) with anti-CLE mAbs (AbCLE) by noncovalent force. The cLFIA completed the entire detection process from de novo to detection results within 30 min thanks to the easy availability and ideal marking efficiency (≤1 min, saving 0.4-10 h) of GC. Encouragingly, the proposed ultrafast GC-ABPS-based cLFIA has also exhibited high sensitivity (0.411 ng mL-1) and low cost (300 times) compared with other cLFIAs. Also, the feasibility of the proposed cLFIA was demonstrated by detecting CLE in beef, pork ham, and skim milk. Finally, the proposed GC-ABPS-based cLFIA has broadened the application range of dyestuffs and provided an effective reference strategy for the application of dyestuffs in food safety monitoring.

Recent advances in meat freshness "magnifier": fluorescence sensing.

To address the serious waste of meat resources and food safety problems caused by the decrease in meat freshness due to the action of microorganisms and enzymes, a low-cost, time-saving and high-efficiency freshness monitoring method is urgently needed. Fluorescence sensing could act as a "magnifier" for meat freshness monitoring due to its ability to sense characteristic signal produced by meat spoilage. Here, the magnification mechanism of meat freshness via sensing the water activity, adenosine triphosphate, hydrogen ion, total volatile basic nitrogen, hydrogen sulfide, bioamines was comprehensively analyzed. The existing "magnifier" forms including paper chips, films, labels, arrays, probes, and hydrogels as well as the application in livestock, poultry and aquatic meat freshness monitoring were reviewed. Future research directions involving innovation of principles, visualization and quantification capabilities for various meats freshness were provided. By critically evaluating the potential and limitations, efficient and reliable meat freshness monitoring strategies wish to be developed for the post-epidemic era.

Next-generation meat preservation: integrating nano-natural substances to tackle hurdles and opportunities.

The increasing global meat demand raises concerns regarding the spoilage of meat caused by microbial invasion and oxidative decomposition. Natural substances, as a gift from nature to humanity, possess broad-spectrum bioactivity and have been utilized for meat preservation. However, their limited stability, solubility, and availability hinder their further development. To address this predicament, advanced organic nanocarriers provide an effective shelter for the formation of nano-natural substances (NNS). This review comprehensively presents various natural substances derived from plants, animals, and microorganisms, along with the challenges they face. Subsequently, the potential of organic nanocarriers is explored, highlighting their distinct features and applicability, in addressing these challenges. The review methodically examines the application of NNS in meat preservation, with a focus on their pathways of action and preservation mechanisms. Furthermore, the outlook and future trends for NNS applications in meat preservation are concluded. The theory and practice summary of NNS is expected to serve as a catalyst for advancements that enhance meat security, promote human health, and contribute to sustainable development.

Diversified organic nanocarriers conquer the limitations of natural substancesNNS based on organic nanocarriers are a reliable and health-promoting optionNNS can manifest their effectiveness through diverse pathways and mechanismsThe utilization of NNS in meat preservation represents a transformative strategy.

COVID-19-inspired "artificial virus" to combat drug-resistant bacteria by membrane-intercalation- photothermal-photodynamic multistage effects.

COVID-19 threatens human life because of the super destructiveness produced from its coronal morphology and strong transmembrane infection based on spike glycoprotein. Inspired by the coronal morphology of COVID-19 and its means of infecting, we designed an "artificial virus" with coronal morphology based on the concept of "defeating superbacteria with superviruses" by self-assembling a transacting activator of transduction peptide with triple-shell porous graphitic carbon nitride (g-C3N4) embedded with cobalt nanoparticles to forcefully infect methicillin-resistant Staphylococcus aureus (MRSA). The results confirmed that this "artificial virus" had unique properties of crossing the bacterial cell membrane barrier, heating the internal bacterial microenvironment and triggering ROS outbreak, based on its coronal morphology, membrane penetration, temperature-rising and heat insulation, oxidase-like activity and excellent visible-light harvesting properties. It had a high sterilization efficiency of 99.99% at 20 min, which was 18.6 times that of g-C3N4, and the efficiency remained at 99.99% after 3 rounds of recycling and reuse. Additionally, it can rapidly inactivate bacteria in river water and accelerate wound healing.

Mussel-Inspired Fabrication of Konjac Glucomannan/Poly (Lactic Acid) Cryogels with Enhanced Thermal and Mechanical Properties.

Three-dimensional nanofibers cryogels (NFCs) with both thermally-tolerant and mechanically-robust properties have potential for wide application in biomedical or food areas; however, creating such NFCs has proven to be extremely challenging. In this study, konjac glucomannan (KGM)/poly (lactic acid) (PLA)-based novel NFCs were prepared by the incorporation of the mussel-inspired protein polydopamine (PDA) via a facile and environmentally-friendly electrospinning and freeze-shaping technique. The obtained KGM/PLA/PDA (KPP) NFCs were characterized by field emission scanning electron microscopy (FE-SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC) and compressive and tensile test. The results showed that the hierarchical cellular structure and physicochemical properties of KPP NFCs were dependent on the incorporation of PDA content. Moreover, the strong intermolecular hydrogen bond interactions among KGM, PLA and PDA also gave KPP NFCs high thermostability and mechanically-robust properties. Thus, this study developed a simple approach to fabricate multifunctional NFCs with significant potential for biomedical or food application.

Recyclable bactericidal packaging films for emperor banana preservation.

Food safety issues and food waste have always been hot topics of concern. This study aimed to develop a recyclable bactericidal packaging film that combines polylactic acid (PLA), graphitic carbon nitride (CN) and carbon nanotubes (CNT) to extend food shelf life. This film exhibited compactness and thermostability, as observed by scanning electron microscope and differential scanning calorimeter. The temperature of P/CN/CNT film could still reach 54 ± 4 °C after being used for 3 times. The film still has bactericidal activity on the 5th cycle use except for L. monocytogenes revealed by morphological characterization on bacteria. This film effectively extended the shelf life of banana to 7 days, as confirmed by measurements of hardness, pH value and total bacterial count of banana. This study provides a packaging film with recyclable bactericidal ability.

Efficient Fresh Lamp Light-Harvesting Films with the Self-Activating Continuous and Recyclable Bactericidal Ability for Ultrapersistent Freshness of Perishable Muscle Food.

A large quantity of perishable muscle food is being wasted due to harmful bacteria infestation during the sales and circulation each year and facing challenges. In this study, a self-activated bactericidal active film (PLA/g-C3N4@PCN-224) responsive to fresh lamp light was prepared, which showed excellent hydrophobicity, water vapor resistance, and thermal stability. Due to the synergistic effect between light-induced reactive oxygen species and the high specific surface area of g-C3N4@PCN-224, this film still maintains 99.99% bactericidal efficacy against Escherichia coli and Staphylococcus aureus after 10 days of continuous bactericidal activity test. The results of cell and hemolysis experiments indicated that the film was safe and nontoxic and can effectively preserve fresh pork for 7 days. Moreover, the film also exhibited a recyclable and efficient killing activity. A strategy for achieving ultrapersistent freshness of perishable muscle food was provided.

Chitosan-based packaging films with antibacterial-sterilization integrated continuous activity for extending the shelf life of perishable foods.

The current food packaging films can be preservative but lack the function of combining antibacterial and sterilization which lead to films can not maximize prolong shelf life of perishable foods. This study provided a new strategy to realize prolonging shelf life of perishable foods by integrating antibacterial and sterilization which focused on applying photodynamic inactivation to films with continuous activity, where curcumin (CUR) and sodium copper chlorophyll (SCC) were loaded into chitosan (CS) films. Compared to pure CS films, the barrier capacity (oxygen permeability and water vapor permeability) and mechanical properties of composite films were improved by introducing CUR and SCC. In addition, the composite film can effectively against food-borne pathogenic bacteria and significantly prolong the shelf life of cherries and pork. The provided strategy has potential application prospects in food preservation packaging.

Multifunctional packaging film with sustained release behavior triggered by pH microenvironment for efficient preservation of pork.

To tackle microbial contamination and spoilage during pork storage, transportation, and sales, a novel packaging film with exogenously driven controlled release behavior was designed through the demand-directed preparation of the emulsion of shellac-encapsulated cinnamaldehyde nano-capsules (SNCs) and the ingenious integration of chitosan (CS) film matrix. Among them, the SNCs were synthesized using the solvent exchange method, allowing controlled release of cinnamaldehyde (CA) in response to volatile alkaline substances present in the meat. The electrostatic interaction between SNCs and CS molecules improved the thermal stability, water resistance, tensile strength, and viscosity of the film-forming solution, while the CA in SNCs enhanced UV resistance and antimicrobial properties of the film. Notably, the CS film with SNC-100 loaded (CSNCs-100 film) achieved 99% inhibition against both E. coli and S. aureus, regulated environmental moisture, and prolonged the freshness duration of pork by an impressive six days.

Characterization and interactions of spoilage of Pseudomonas fragi C6 and Brochothrix thermosphacta S5 in chilled pork based on LC-MS/MS and screening of potential spoilage biomarkers.

Pseudomonas and Brochothrix are the main spoilage organisms in pork, and each of these plays an essential role in the spoilage process. However, the effect of co-contamination of these two organisms in pork has not been elucidated. The changing bacterial communities during spontaneous spoilage of pork at 4 °C were evaluated using high-throughput sequencing. The dominant spoilage bacteria were isolated and these were identified as Pseudomonas fragi C6 and Brochothrix thermosphacta S5. Chilled pork was then experimentally contaminated with these strains, individually and in combination, and the progression of spoilage was assessed by analyzing various physicochemical indicators. These included total viable counts (TVC), pH, color, total volatile basic nitrogen (TVB-N), and detection of microbial metabolites. After 7 days of chilled storage, co-contaminated pork produced higher TVC and TVB-N values than mono-contaminated samples. Metabolomic analysis identified a total of 8,084 metabolites in all three groups combined. Differential metabolites were identified, which were involved in 38 metabolic pathways. Among these pathways, the biosynthesis of alkaloids derived from purine and histidine was identified as an important pathway related to spoilage. Specifically, histidine, histamine, AMP, IMP, GMP, succinic acid, and oxoglutaric acid were identified as potential spoilage biomarkers. The study showed that the combined presence of P. fragi C6 and B. thermosphacta S5 bacteria makes chilled pork more prone to spoilage, compared to their individual presence. This study provides insights that can assist in applying appropriate techniques to maintain quality and safety changes in meat during storage and further the assessment of freshness.

Natural melanin nanoparticle-based photothermal film for edible antibacterial food packaging.

Although nanocomposite films had shown excellent potential in antibacterial food packaging, their potential harmful impact limits their further application in reality. Therefore, exploring a Generally Recognized As Safe (GRAS) nanomaterial that has antibacterial ability is the pioneer for the fabrication of a real edible nanocomposite-based antibacterial packaging film. Herein, for the first time by using the natural nanostructure extracted from cuttlefish ink, an edible antibacterial food packaging with high safety were constructed. The natural melanin nanoparticles (NMPs) in cuttlefish ink have good photothermal conversion ability. As such, by incorporating with natural pectin (PC) film and with near infrared (NIR) irradiation triggering, the results show that PC/NMPs films perform high-efficiency and short-term bactericidal activity against foodborne pathogenic bacteria, including thermotolerant Listeria monocytogenes. The sterilization rate could reach more than 90 % within only 5 min. Also, this nanocomposite film showed better mechanical properties, thermal stability and barrier properties than the neat pectin film. Antibacterial test on food sample also proved the good antibacterial ability of the PC/NMPs films. Therefore, exploring GRAS natural functional nanocomposite film is expected to be the effective way to promote edible nano-antibacterial packaging.

Konjac glucomannan films with quasi-pasteurization function for tangerines preservation.

This study presents functional KCNGO films obtained by incorporating graphite carbon nitride (g-C3N4) and graphene oxide (GO) into the konjac glucomannan (KGM) matrix. The concept of quasi-pasteurization proposed in current work refers to the shorter time needed and lower sterilization temperature used than that of the conventional pasteurization. The films had a compact microstructure formed by the hydrogen bond interactions, as revealed by morphology characterization, FTIR and rheological analysis. It also had excellent hydrophobicity, thermal stability, mechanical properties and water vapor barrier ability. Notably, this film exhibited potent bactericidal activity against foodborne pathogens based on functions of fast heating and instantaneous low temperature bactericidal (4 min, 55 ± 1 °C). The quasi-pasteurization efficacy and mechanism of films were explored. Moreover, this film was safe and can keep the "fresh-like" characteristics of the tangerines for 24 days. Therefore, a quasi-pasteurization strategy is provided in food preservation field.

Enhanced functional properties of chitosan films incorporated with curcumin-loaded hollow graphitic carbon nitride nanoparticles for bananas preservation.

The exploration of novel functional packaging films is of great scientific and technological interest. Herein, a novel chitosan/hollow g-C3N4/curcumin (CS-HCNS-Cur) biocomposite films was successful fabricated with integrated functions of slow release, antimicrobial activity and food freshness preservation. CS-HCNS-Cur films take the advantages of the excellent thermal stability and slow-release ability of HCNS to curcumin. Among the characterizations including scanning electron microscopy, transmission electron microscope, atomic force microscopy, fourier transform infrared spectroscopy, mechanical properties and the rheological properties measurements confirmed the successful fabrication of CS-HCNS-Cur films. The averaged water contact angle and water vapor permeability of this film were 105.83° and 105.03 × 10-5 g·mm (m2·h·kPa)-1, respectively. This film showed pH-responsive and slow-release ability. Moreover, this film can effectively store bananas for 10 days. Therefore, CS-HCNS-Cur films have promising potential for applications in functional food packaging.

Hydrogel loading 2D montmorillonite exfoliated by anti-inflammatory Lycium barbarum L. polysaccharides for advanced wound dressing.

Designing wound dressing materials with hemocompatibility, suitable mechanical properties, outstanding hemostatic effects and anti-inflammatory activity is of great practical significance for wound management. Herein, a hemostatic hydrogel loaded with Lycium barbarum L. polysaccharide (LBP)-functionalized ultrathin MMT nanosheets (L-MMT NSs) was fabricated for efficient hemostasis and wound healing. Loading the L-MMT NSs into polyvinyl alcohol (PVA), the obtained P-L-MMT hydrogel exhibited a 3D porous structure with good swelling properties, cytocompatibility, hemocompatibility, and anti-inflammatory activity. Importantly, in vivo investigations demonstrated that the P-L-MMT hydrogel exerts outstanding hemostasis activity in the hemorrhaging mouse liver model and reduces tissue damage caused by inflammation to shorten wound healing time. Altogether, the convenient exfoliation and functionalization of bulk MMT using LBPs make this inexpensive and rising nanostructure more attractive in the application of nanomedicine. Moreover, due to the synergy between hemostasis and anti-inflammation, this newly developed multifunctional P-L-MMT hydrogel represents a promising material in biomedical fields.

Advanced Coatings with Antioxidant and Antibacterial Activity for Kumquat Preservation.

An active coating is one of the best ways to maintain the good quality and sensory properties of fruits. A new active coating was prepared by incorporating curcumin and phloretin into the konjac glucomannan matrix (KGM-Cur-Phl). The fourier infrared spectroscopy, rheology and differential scanning calorimetry confirmed the successful fabrication of this coating. This coating showed excellent antioxidant activity revealed by the 95.03% of ABTS radicals scavenging ratio and 99.67% of DPPH radicals scavenging ratio. The result of bacteria growth curves showed that it could effectively inhibit the growth of Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Salmonella typhimurium. The results of firmness, titratable acid and pH showed that it effectively prolonged the shelf life of kumquat. A novel konjac glucomannan-based active coating was provided.

Efficient and reusable photocatalytic river water disinfection by addictive graphitic carbon nitride/magnesium oxide nano-onions with particular "nano-magnifying glass effect".

Photocatalytic disinfection is a promising way to combat bacterial pollution in the water environment. Inefficient use of visible light and undirected diffusion of reactive oxygen species (ROS) reduce photocatalytic disinfection efficiency. Herein, inspired by the concentrating effect of convex lens, photocatalysts with particular "nano-magnifying glass effect" (TCNMgNOs) were designed by embedding magnesium oxide with "converge effect" into the tailored hierarchical triple-shell porous g-C3N4 with "one light multi-purpose effect" to boost the visible-light utilization. Meanwhile, the ATPase hydrolysis homeostasis of bacteria was destroyed by TCNMgNOs to achieve the targeted movement of ROS. The results confirmed that the photocatalytic sterilization efficiency of TCNMgNOs was amplified by 30 times over g-C3N4, which was achieved by focusing visible light, multiple reflecting visible light and light transmission within the porous thin shells as well as the "addictive sterilization mechanism". The sterilization efficiency still maintains 98.8 % (15 min) after 6 rounds recycling and reusing in practical river water disinfection. A novel pathway for fighting against microbial contaminants in natural water was explored.

Enhanced antimicrobial activity of konjac glucomannan nanocomposite films for food packaging.

This paper aims at providing a new strategy for developing konjac glucomannan-based antibacterial films with excellent performances. Here, novel nanocomposite films based on photodynamic and photothermal synergism strategy were developed by incorporating graphite carbon nitride nanosheets/MoS2 nanodots (CNMo) into konjac glucomannan (KGM) matrix. Scanning electron microscope, transmission electron microscope, high resolution transmission, high angle annular dark field and element mapping confirmed the successful fabrication of CNMo. The steady and dynamic rheological behavior as well as the good stability of film-forming solution showed that the intermolecular hydrogen bonding was formed. The influences of CNMo content on the structural, mechanical and thermal properties as well as hydrophobicity of KGM films were investigated. This film has a broad-spectrum antibacterial activity. It could prolong the shelf life of cherry tomatoes. Moreover, hemolysis and cells experiment confirm that this film is safe. This strategy is expected to broaden the application of antibacterial packaging.

Visible light responsive, self-activated bionanocomposite films with sustained antimicrobial activity for food packaging.

The research on a new type of low-cost, less-loss and adjustable sustained antibacterial activity food packaging films with self-activation ability and great industrialization potentiality is of great scientific and technological interest. Herein, a novel chitosan/negatively charged graphitic carbon nitride self-activation bionanocomposite films was prepared by one-step electrostatic self-assembly. First, the antibacterial efficiency of this film could reach to 99.8 ± 0.26% against E. coli and 99.9 ± 0.04% against S. aureus through self-activated under visible light. Second, this film can effectively extend the shelf life of tangerines to 24 days. Hemolysis and cell experiment test proved that this film was safe and nontoxic. Finally, negatively charged graphitic carbon nitride with low-cost can improve the mechanical, thermal and hydrophobic properties of neat chitosan films. This work can provide a new pathway for the preparation of low-cost packaging films with excellent visible light responsive property and sustainable antibacterial activity.