Rapid and field-deployable detection of foodborne pathogens remains a critical challenge for ensuring food safety and public health, particularly in resource-limited settings. Herein, we firstly report a vibration-enhanced recombinase polymerase amplification (RPA) platform based on a foldable multilayer microfluidic paper-based analytical device (mu PAD) for on-site detection of Vibrio parahaemolyticus (V. parahaemolyticus). Unlike conventional paper-based amplification systems that suffer from diffusion limitations within porous matrices, the proposed platform introduces a low-power micro-vibration module to enhance reagent mixing and improve amplification efficiency. The proposed system integrates pre-stored, air-dried RPA reagents, a portable heating unit with vibration-assisted mixing, and a smartphone-based fluorescence readout module. Isothermal amplification is initiated directly on the paper substrate by simple sample loading, enabling nucleic acid amplification and fluorescence detection within 20 min at 40 degrees C. The assay achieves a detection limit of 102 CFU/mL, comparable to that obtained using a laboratory fluorescence spectrometer. The platform exhibits high specificity and reliable performance in spiked seawater and seafood samples, with results consistent with standard culture-based methods. By addressing mass-transfer limitations in paper-based amplification and enabling system-level integration for portable detection, this platform provides a practical and scalable solution for on-site food safety monitoring.

To address the emerging multidrug-resistance crisis caused by Klebsiella pneumoniae, we expressed the endolysin Lys59 derived from phage VB_KpP_HS106 and performed a comprehensive analysis of its antibacterial activity and structural features. Molecular modeling revealed that Lys59 carries a highly positively charged N-terminus and an amphipathic helix at the C-terminus. In vitro antibacterial assays showed that Lys59 exhibited significant bactericidal activity against K. pneumoniae with an approximately 4 log reduction at 50 µg/mL in 2 h. Meanwhile, Lys59 exhibited potent, broad-spectrum activity against both Gram-negative and Gram-positive bacteria. Stability analysis indicated that Lys59 retained high activity over a pH range of 3-9 and a temperature range of 4-55 °C. Notably, the antibacterial activity of Lys59 was found to be regulated by metal ions. Molecular docking indicated that K+ can enhance binding stability by interacting with ASN35 and VAL57. In contrast, Mg2+ and Ca2+ suppressed catalytic function by binding to the essential GLU17 residue. Furthermore, treatment with 200 µg/mL of Lys59 resulted in a 44.6% reduction in K. pneumoniae biofilm biomass. Overall, this study identified a phage-derived endolysin with broad-spectrum antimicrobial activity and demonstrated its potential as an antibacterial agent against multidrug-resistant K. pneumoniae.

The food industry is currently facing two critical challenges: food waste caused by inadequate protection of traditional packaging, and environmental pollution induced by the poor degradability of petroleum-based packaging materials. Against this backdrop, the development of sustainable and multifunctional packaging materials has become an inevitable trend for industrial development. Derived from renewable resources, bio-based aerogels have attracted extensive attention due to their excellent properties such as high specific surface area, ultra-low density, high porosity, and biodegradability, emerging as a promising sustainable alternative to traditional packaging materials. This paper systematically reviews the raw material sources, fabrication methods, functionalization strategies and practical applications of bio-based aerogels in the field of food packaging, highlights the modification techniques tailored to the specific requirements of food packaging, and summarizes the latest application achievements of various techniques in diverse food packaging scenarios. Studies have shown that bio-based aerogels exhibit great application potential in food cushioning protection, active preservation and intelligent monitoring of food freshness. Their multiple functions can effectively prolong the shelf life of food and realize real-time monitoring of food freshness, making them an ideal sustainable material to replace traditional petroleum-based plastic packaging. Future research should prioritize the development of scalable fabrication processes and cost-effective production strategies to accelerate the industrial translation and commercialization of bio-based aerogels.

This study simultaneously synthesized three plant-derived zinc oxide nanoparticles (ZnO NPs-L, ZnO NPs-P, ZnO NPs-M) using loquat, perilla, and mulberry leaf extracts as reducing and capping agents. The results indicated that all three nanoparticles exhibited the typical hexagonal wurtzite structure with intact crystal lattices and high crystallinity. Characteristic ZnO absorption peaks appeared within the 370-380 nm range in the UV-Vis spectra, confirming successful nanoparticle formation. DLS and zeta potential analyses revealed uniform particle size distributions and negatively charged surfaces, indicating excellent colloidal stability. Antibacterial assays demonstrated inhibitory effects of all three ZnO NPs against Escherichia coli and Staphylococcus aureus, following the activity order ZnO NPs-L>ZnO NPs-P>ZnO NPs-M. DPPH· and ABTS·+ radical scavenging assays revealed antioxidant activity trends consistent with antibacterial performance. These findings demonstrate that edible leaf extract-mediated ZnO NPs represent promising green nanomaterials with potential applications in active food packaging.

Endolysin is an enzyme produced by bacteriophages during the lysis cycle, capable of specifically degrading peptidoglycan in the bacterial cell wall. However, its application against gram-negative bacteria is limited by the outer membrane (OM) barrier, which prevents access to the cell wall. In this study, the endolysin Lys53 from the phage vB_VpaS_1601 was modified by fusing cationic peptides to obtain the engineered protein Lys53-C5aa. Lys53-C5aa significantly reduced Vibrio parahaemolyticus by 3.51 log CFU/mL at a concentration of 6.4 μM within 30 min and exhibited a wider lysis spectrum. Lys53-C5aa exhibited optimal lytic activity within a temperature range of 4 to 65 °C and a pH range of 5 to 10. Lys53-C5aa significantly enhanced OM permeability of V. parahaemolyticus and led to the leakage of intracellular nucleic acids, reaching 110.5 ng/μL at 75 min. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) revealed that Lys53-C5aa disrupted the structural integrity of V. parahaemolyticus. Lys53-C5aa also showed removal (41.83%) and inhibition (61.01%) effects on the biofilm of V. parahaemolyticus. Lys53-C5aa reduced V. parahaemolyticus counts in oysters by 2.14 and 2.40 log CFU/g at 4 °C and 25 °C, respectively, while causing reductions of 2.21 and 2.69 log CFU/g in Pacific white shrimp under the same conditions. These findings indicate that Lys53-C5aa represents a highly promising candidate for controlling V. parahaemolyticus contamination in aquatic products.

In this study, the inhibitory mechanism of chlorogenic acid-grafted chitosan (CA-g-CS) against Pseudomonas fluorescens at the gene level were investigated by combining transcriptomics and bioinformatics. Transcriptome results demonstrated that differential expression genes (DEGs) in the CA-g-CS treated group were concentrated in inorganic ion transport metabolism and outer membrane. Protein-protein interaction (PPI) network analysis showed that a large number of significantly down-regulated genes were closely related to iron transport in bacteria. Among them, Tonb-dependent outer membrane protein receptors Fiu, TbpA and FecA of P. fluorescens were significantly down-regulated under CA-g-CS treatment by -5.092, -3.768 and - 3.716 Log2 Fold change. Subsequently, atomic force microscope (AFM) and Fourier transform infrared spectroscopy (FT-IR) indicated that CA-g-CS treatment causes membrane surface collapse and reduces the bacterial membrane protein content of P. fluorescens. Moreover, CA-g-CS treatment significantly reduced the adsorption of Fe3+ by P. fluorescens, which decreased the iron (Fe) uptake and intracellular iron (Fe) content by 27.27 % and 37.08 %, respectively, and retarded the growth activity of strain in iron-limiting medium. Overall, CA-g-CS could kill P. fluorescens by inhibiting the transcription of membrane channel proteins and inhibit the transport and uptake of iron (Fe).

Synergistic antibacterial effect is a promising way to overcome the challenge of drug-resistant microbial contamination in food. In this study, carnosic acid/caffeic acid (CR/CF) combination showed a stronger synergistic antibacterial effect on broad-spectrum multidrug-resistant Staphylococcus aureus (MRSA) than methicillinsensitive Staphylococcus aureus ATCC29213. The co-treatment also reduced the biofilm formation by 60.7-80 % and the structure of MRSA 009 was destroyed and loosed. Metabolic activity of biofilm cells was inhibited by 37.9 %-68.9 %. The exopolysaccharides and extracellular proteins were inhibited by 73.3 % and 42.7 %, respectively. Furthermore, CR/CF combinations at 1 MIC +1/2 MIC reduced about 2.4 log CFU/cm2 of MRSA on lettuce within 15 min, which had a similar effect as 0.2 % (v/v) sodium hypochlorite (NaOCl). Thus, CR/CF combination may provide a promising option to tackle antimicrobial resistance and limit MRSA spread in the fresh produce.

This study aimed to investigate antimicrobial effects of caffeic acid (CA) against multidrug-resistant Vibrio parahaemolyticus and Vibrio cholerae both in vitro and in shrimp (Penaeus vannamei). CA at concentrations of 0.47-0.94 mg/mL disrupted bacterial membrane integrity, resulting in increased nucleic acid leakage and decreased intracellular DNA content. At 1 × minimum inhibitory concentration (MIC), CA inhibited biofilm formation, with maximum reductions of 76.4 % and 68.3 % for V. parahaemolyticus and V. cholerae, respectively. Molecular dynamics simulations revealed stable binding between CA and critical bacterial proteins except for ompW, with binding energies of -6.6 to -5.8 kcal/mol. RT-qPCR further confirmed downregulation of related gene expression. In shrimp, CA treatment (6 × MIC, 4 h) reduced both Vibrio species by >4.4 log CFU/g. These results demonstrate the potent anti-Vibrio activity of CA, underscoring its potential as a plant-derived antimicrobial agent for ensuring microbial safety in seafood processing industry.

Staphylococcus aureus and Klebsiella pneumoniae are significant and prevalent pathogens associated with bovine mastitis on dairy farms worldwide, resulting in severe infections in both dairy cows and, subsequently, human beings. Fast and dependable pathogen diagnostics are essential to minimize the effects of cow mastitis and human infections. The aim of this research was to develop a duplex recombinase-aided amplification (RAA) combined with the lateral flow dipstick (LFD) method, which was used for rapid, simultaneous detection of S. aureus and K. pneumoniae. The SKII culture medium for S. aureus and K. pneumoniae cocultivation was developed in this study. By optimizing the duplex RAA–LFD reaction conditions in terms of primer concentration, amplification temperature, and reaction time, the duplex RAA–LFD assay could successfully detect S. aureus and K. pneumoniae when the reaction was conducted at 39 °C for 20 min. The duplex RAA–LFD method demonstrated good specificity, exhibiting no cross-reactivity with other pathogens. In addition, the detection limit of the duplex RAA–LFD for S. aureus and K. pneumoniae was 60 fg of genomic DNA and 1.78 × 103 and 2.46 × 103 CFU/mL of bacteria in pure culture. Moreover, the duplex RAA–LFD technique is capable of identifying S. aureus and K. pneumoniae in artificially spiked milk samples even at very low initial concentrations of 1.78 × 101 and 2.46 × 100 CFU/mL, respectively, after 6 h of enrichment. The result of the actual samples showed that the total concordance rate of the duplex RAA–LFD method with the biochemical identification method and PCR method could reach 92.98~98.25% with high consistency. The results of this study indicated that the duplex RAA–LFD assay, which is a precise, sensitive, and simple field testing technique, can be used to identify S. aureus and K. pneumoniae and is expected to be used for disease diagnosis.

The rising prevalence of drug-resistant bacterial infections poses a significant public health challenge. This is particularly evident in Klebsiella pneumoniae, a pathogen that can cause severe diseases including pneumonia, liver abscesses, sepsis, and death. Phage therapy, which involves the use of active phages or endolysins, is gaining attention as an alternative to antibiotics in the management of infections. A lytic bacteriophage (vB_KpP_HS37) targeting K. pneumoniae from sewage at Shanghai No.6 People's Hospital was isolated in this study. The phage exhibited robust activity across a temperature range of 4 to 50 °C and pH values from 5.0 to 10.0. Genome sequencing revealed that HS37 possesses a linear double-stranded DNA genome of 74,084 base pairs with a GC content of 45.13 %. It is a new species of the Sugarlandvirus genus, Demerecviridae family. The HS37 endolysin (Lys41) was successfully expressed in vitro and subsequently characterized. Notably, the lytic spectrum of Lys41 was wider compared to its parental phage, HS37. Lys41 exhibited optimal lytic activity within a temperature range of 37 to 55 °C and a pH range of 4.0 to 6.0. When combined with EDTA and KCl, Lys41 effectively lysed K. pneumoniae and showed a more extensive lytic spectrum than phage HS37. The combination of Lys41 and 15 ppm potassium sorbate achieved reductions of 1.6 and 0.9 Log10 CFU/mL in K. pneumoniae on lettuce and cooked chicken, respectively. These findings demonstrate the efficacy of phage HS37 and its endolysin Lys41 as a therapeutic agent against K. pneumoniae infections.

VtrB is a novel ToxR-like transcriptional regulatory protein located on the Vibrio parahaemolyticus pathogenicity island (Vp-PAI). It can be induced by bile to activate the Vp-PAI genes, thereby enhancing the cytotoxicity and enterotoxicity of V. parahaemolyticus. However, the molecular mechanism of VtrB in bile resistance has not been thoroughly examined. The purpose of this study was to investigate the global regulatory network of VtrB. V. parahaemolyticus strain with vtrB deletion (ΔvtrB) was established. The response to bile of the wild type and ΔvtrB strain suggested that VtrB contributed to bile resistance and supplied a survival advantage to V. parahaemolyticus in the presence of bile. Motility assay and static biofilm assay showed that VtrB negatively regulates V. parahaemolyticus motility and biofilm formation in a bile-dependent manner. The cytotoxicity assay showed that ΔvtrB exhibited a significant reduction in toxicity toward Caco-2 cells. RNA sequencing and qRT-PCR were performed, and functions of the differentially expressed genes (DEGs) were investigated. RNA sequencing identified 701 DEGs in the ΔvtrB strain, including 462 up-regulated genes and 239 down-regulated genes. Functional enrichment analysis of DEGs indicated that VtrB contributes to bile resistance of V. parahaemolyticus by altering genes related to "Bacterial secretion system", "Flagella assembly and chemotaxis", "Biofilm formation", "Two-component system", "Quorum sensing", "Glyoxylate and dicarboxylate metabolism", "Citrate cycle", and "Carbon metabolism". Our findings suggested that VtrB may be part of a complex transcriptional network that regulates the expression of virulence genes and multiple cellular phenotypes.

Microbiological contamination remains a significant challenge on the spoilage of aquatic products. The rapid proliferation of specific spoilage organisms (SSOs) in aquatic products can lead to the generation of harmful substances, thereby degrading the quality of these products. This underscores an urgent demand for innovative strategies to overcome this challenge. Quorum sensing inhibitors (QSIs) emerge as a promising approach in that they can mitigate microbial contamination by disrupting bacterial communication mechanisms, especially in strains prevalent in aquatic products. The signaling molecules involved in quorum sensing (QS) in aquatic products play a critical role in regulating bacterial population behavior. Inhibiting the QS system of bacteria offers an innovative strategy for bacterial control, which does not rely on the direct killing of bacteria but rather on the attenuation of their spoilage effects in aquatic products by disrupting their collective behavior. Plant extracts employed as QSIs have demonstrated the potential to effectively prolong the shelf life of food products owing to their natural, environmentally friendly, and potentially antimicrobial properties, thus rendering them a promising solution for the preservation of aquatic products. This comprehensive review provides an in-depth analysis of common signaling molecules and QS systems in SSOs of aquatic products. Besides, it evaluates the potential of plant extracts as QSIs to prolong the shelf life of aquatic products by inhibiting their QS mechanisms. Additionally, this review also highlights the potential application of plant-derived QSIs in the preservation of aquatic products.

This study firstly elucidates the molecular mechanism by which chlorogenic acid-grafted chitosan (CA-g-CS) inhibits the spoilage activity of Pseudomonas fluorescens from the iron-uptake perspective. By constructing deletion mutants of iron uptake genes in P. fluorescens and combining in vitro functional validation with a refrigerated sterile fish fillet spoilage model, we elucidated the functional roles of these genes and the mechanism of CA-g-CS in inhibiting P. fluorescens spoilage. Results indicate that Δfiu, ΔfecA, and ΔefeO significantly reduce iron uptake under Fe-limited conditions. The Δfiu and ΔfecA mutants demonstrate impaired Fe3+ adsorption function, while ΔefeO mutants show decreased Fe2+ uptake capacity. All mutants exhibited decreased intracellular iron and biofilm formation, with Δfiu defects being most severe. Overexpression of fiu (WT::fiu) enhances iron uptake in the strain, and this effect is strongly inhibited by CA-g-CS. Confocal laser scanning microscopy (CLSM) and molecular docking indicated that CA-g-CS acts extracellularly, binding to Fiu via hydrogen bonding and hydrophobic interactions. In the sterile fish fillet model, the Δfiu mutant exhibited reduced growth rate, slowed lipid oxidation, and delayed protein degradation, thereby decreasing the spoilage rate of the fillets. In summary, the fiu, fecA and efeO genes synergistically regulate iron uptake in P. fluorescens, while CA-g-CS blocks iron uptake by specifically binding to the iron transporter Fiu, thereby inhibiting the strain's spoilage activity. This study provides a novel theoretical foundation for targeting the iron-uptake pathway as a strategy to inhibit bacterial spoilage.

Bacteriophages have reemerged to potentially replace or complement the role of antibiotics, as bacterial viruses have the ability to inactivate pathogens. However, certain intrinsic limitations of phages overshadow their clinical application, particularly their narrow host spectrum and rapid development of resistance upon treatment. This study aimed to explore the synergistic antimicrobial effect of phage combined with antibiotics against Klebsiella pneumoniae. The time-killing experiments in vitro showed that phage and gentamicin combination displayed synergistic bactericidal activity, leading to a reduction in the minimum inhibitory concentration of gentamicin. Furthermore, the phage HS106/gentamicin combination significantly inhibited biofilm formation and eliminated mature biofilms. On the other hand, phage treatment for 2 h before gentamicin treatment produced better synergistic inhibitory effect. The use of phage followed by gentamicin can effectively inhibit the efflux effect. Surprisingly, the phage HS106/gentamicin combination still exhibited antimicrobial activity against phage-resistant mutants and double-resistant mutants. Finally, the phage HS106/gentamicin combination significantly increased the survival rate of zebrafish infected with K. pneumoniae, indicating its excellent bactericidal activity in vivo. Overall, the phage HS106/gentamicin combination may provide a promising approach for treating infections caused by high-level multidrug-resistant K. pneumoniae.

In this study, we isolated 214 Vibrio cholerae strains from aquatic (shrimp, crab, grass carp, and crucian carp) and their cultured environment in Shanghai, China. The virulence, serotype, and antimicrobial susceptibility were tested, and polymerase chain reaction (PCR) was used to detect antimicrobial resistance genes. Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) was employed for cluster analysis of the isolated strains. The results showed that V. cholerae was found in 47.9% (114/238) of aquatic samples, with the highest detection rate in shrimp (81.1%), and the detection rate was highest in summer (70.0%). Most of the strains were non-O1/O139 groups, and virulence genes rtxC and hap had the highest detection rates of 92.5% and 91.1%. Of the 214 isolates, 69.6% were multidrug-resistant (MDR). The resistance rate of V. cholerae to sulfamethoxazole, ampicillin, and erythromycin was 97.2%, 85.5%, and 70.1%, and that to imipenem, tetracyclines, and aminoglycosides was less than 5%. The MAR index ranged from 0.05 to 0.47. When V. cholerae was screened for antimicrobial resistance genes, β-lactams CARB, chloramphenicol floR, and sulfonamides sul2 were found in 19.6%, 7.9%, and 6.5% of isolates, respectively. The results of ERIC-PCR clustering showed that the isolates had a high degree of genetic diversity. The widespread distribution of virulent and MDR V. cholerae strains poses a potential threat to food safety and public health, calling for improved monitoring and control measures in the aquaculture industry.

Hypervirulent Klebsiella pneumoniae (hvKP), classified as ESKAPE pathogens, represent substantial global public health threats owing to their antibiotic resistance and virulence factors. Infections caused by hvKP are often associated with high mortality rates and healthcare costs, and the pathogen tends to form biofilms, which further complicates treatment. Thus, novel therapeutic strategies are urgently needed for K. pneumoniae infections. Bacteriophage-derived enzymes, including depolymerases and endolysins, provide a promising alternative antimicrobial strategy by specifically targeting and degrading bacterial cell walls and capsular polysaccharides. However, the outer membrane of Gram-negative bacteria limits the activity of endolysins, and the lack of bactericidal activity in depolymerases further restricts their application. Future development must address these obstacles. Here, we discuss the virulence factors of K. pneumoniae, review the structure and mechanisms of depolymerases and endolysins, and summarize recent research advances in the prevention and treatment of K. pneumoniae infections. Furthermore, based on the current challenges faced by depolymerase and endolysin therapies against K. pneumoniae infections, we propose a novel chimeric protein design that combines the β-helix domain with the catalytic domain of endolysin, or employs the SpyTag/SpyCatcher system to facilitate the recombination of depolymerase and endolysin. This approach aims to enhance their antibacterial and antibiofilm activities, offering promising potential for the development of new antimicrobial agents against K. pneumoniae.

In this study, soy isolate protein / chitooligosaccharide (SPI/COS) glycosylated conjugates was prepared and employed as an emulsifier to stabilize carvacrol-loaded nanoemulsions (CNE-SPI/COS). The antibacterial properties and mechanism of CNE-SPI/COS against S. putrefaciens was investigated. The results of microbial growth curves and confocal laser scanning microscopy (CLSM) results showed that CNE-SPI/COS effectively inhibited the growth of S. putrefaciens and the killing effect of CNE-SPI/COS on S. putrefaciens was concentration-dependent. Field emission scanning electron microscopy (FESEM) images showed that CNE-SPI/COS caused folds, shrinkage, rupture and even lysis of S. putrefaciens. The results showed that CNE-SPI/COS inhibited the growth and reproduction of S. putrefaciens mainly through three targets: (i) the reduction of alkaline phosphatase (AKP) activity and protein leakage indicated that CNE-SPI/COS disrupted the integrity of cell wall and cell membrane; (ii) the reduction of intracellular protein and ATP content indicated that CNE-SPI/COS interfered the synthesis of intracellular nutrient and synthesis of energy-supplying substances; (iii) changes in the activities of succinate dehydrogenase, pyruvate kinase, and glucose 6-phosphate dehydrogenase indicated that CNE-SPI/COS impeded the normal cellular metabolic pathways such as the tricarboxylic acid cycle, the glycolytic pathway, and the pentose phosphate pathway, and the decrease in superoxide dismutase activity indicated that CNE-SPI/COS disrupted the defense system against oxidative stress. In conclusion, the encapsulation of carvacrol into the nanoemulsion system can provide theoretical support and methodological guidance for the application of nanoemulsions in microbial decontamination of aquatic products.

Vibrio parahaemolyticus poses a serious threat to the aquaculture industry and human health. Two novel phages (vB_VpaS_1601 and vB_VpaP_1701) were isolated from oysters. Phage vB_VpaS_1601 and phage vB_VpaP_1701 exhibited short latent periods and high burst sizes. In addition, both phages showed stability across a broad range of temperature and pH conditions. Genomic analyses revealed that phage vB_VpaS_1601 is a novel genus of the Caudoviricetes class, and vB_VpaP_1701 is a new species of the Maculvirus genus, Autographiviridae family. Both single phage and phage cocktail can inhibit V. parahaemolyticus proliferation, with phage cocktail (MOI = 1) can completely inhibit the growth of V. parahaemolyticus within 12 h. The phages reduced biofilm formation by 53.57 % (vB_VpaS_1601) and 64.88 % (vB_VpaP_1701), while the cocktail achieved 46 %-78 % inhibition. Phage cocktail can reduce V. parahaemolyticus by 1.53-2.74 log CFU/cm3 in salmon and 1.56-2.91 log CFU/cm3 in oysters. These results indicated that vB_VpaS_1601 and vB_VpaP_1701 have potential as antibacterial agents for controlling V. parahaemolyticus in seafood products.

Methicillin-resistant Staphylococcus aureus (MRSA), resistant to beta-lactam antibiotics, presents critical treatment challenges. Bacteriophages (phages) are increasingly considered an alternative therapy for combating antibiotic-resistant pathogens. This study aimed to isolate and characterize a novel lytic phage targeting MRSA, evaluate its efficacy in biofilm disruption and potential as a biocontrol agent in milk. Phage vB_SauH_SPJ2 was successfully isolated and exhibited a short latent period (15 min) and high burst size (99 PFU/cell). Stability assays showed that SPJ2 remained stable across a broad range of temperatures (4-50 degrees C) and pH (3-12) conditions. Morphological and genomic analyses revealed that SPJ2 represented a novel species within the Silviavirus genus, belonging to the Twortvirinae subfamily of the Herelleviridae family, lacking resistance genes. SPJ2 significantly disrupted preformed biofilms of MRSA SA008 (59.05 %), MDR S. aureus SA009 (79.15 %), and S. aureus CMCC26003 (56.72 %) after 24 h, while inhibiting biofilm formation by 86.43 %, 44.56 %, and 71.38 % in a concentration-dependent manner, respectively. The SPJ2-encoded lysin may lyse bacteria and degrade extracellular polymeric substances (EPS), thereby inhibiting and removing biofilm synthesis. At a multiplicity of infection (MOI) of 1000, SPJ2 exhibited effective bactericidal activity against S. aureus at 25 degrees C and 4 degrees C. Notably, SPJ2 reduced the concentration of S. aureus in milk to undetectable levels within 6 h at 25 degrees C, demonstrating exceptional biocontrol potential. Statistical significance was analyzed using two-way ANOVA (p < 0.05). These findings highlight the adaptability of SPJ2 to dairy processing environments, efficacy against MRSA, and promise as a synergistic antimicrobial agent for clinical and industrial applications.

Saikosaponin A (SSA), the primary active monomer derived from the Radix bupleuri, demonstrates a diverse array of pharmacological activities, including anti-inflammatory, antitumor, analgesic, anti-fibrotic, antidepressant, and immune-modulating properties. Despite its potential therapeutic impact on various human diseases, comprehensive studies exploring SSA's efficacy in these contexts remain limited. This review synthesizes the current research landscape regarding SSA's therapeutic applications across different diseases, highlighting critical insights to overcome existing limitations and clinical challenges. The findings underscore the importance of further investigations into SSA's mechanisms of action, facilitating the development of targeted therapeutic strategies and their translation into clinical practice.