Search Results (185)

Narrow host specificity and bacterial resistance often limit single-phage treatments. Phage cocktails address these challenges by expanding the host range, reducing resistance, and enhancing bacterial inactivation. This study aimed to develop an optimised phage cocktail targeting Escherichia coli, Salmonella enterica serovar Typhimurium, Salmonella enterica serovar Enteritidis, and Aeromonas hydrophila, key pathogens in bivalve consumption. Twelve phages were isolated, purified, and screened for bacterial inactivation using resazurin-based viability assays. Host range analysis showed that all phages infected at least one additional bacterial species, with four (phEc4, phSE1, phAh2, phAh4) targeting three of the four bacteria. Cocktail formulation aimed to maximise bacterial reduction while balancing host range expansion with factors such as the risks of resistance development and inter-phage competition. Among the tested combinations, the most effective cocktail consisted of E. coli phage phEc3, S. Typhimurium phage phST1, S. Enteritidis phage phSE1, and A. hydrophila phage phAh2. Future studies should evaluate the cocktail’s efficacy in vitro and assess both safety and performance in vivo in bivalve depuration systems. Full article

Osteoarticular infections (OAIs), including osteomyelitis, septic arthritis, prosthetic joint infections, and facture-related infections, remain a major challenge due to biofilm formation and the prevalence of multidrug-resistant (MDR) pathogens. Although OAIs are predominantly caused by Staphylococcus aureus and coagulase-negative staphylococci, the increasing incidence of MDR Gram-negative infections adds further complexity to their management. Standard approaches, combining surgery and prolonged antibiotic therapy, frequently result in recurrence and poor outcomes. Bacteriophage (phage) therapy has emerged as a promising adjunct or alternative approach, offering high host specificity, replication at the infection site, and activity against biofilm-embedded bacteria. This review highlights recent advances in phage therapy for OAIs, focusing on administration routes (intravenous, intra-articular, topical, and oral) and on novel pharmaceutical delivery systems such as hydrogels, bone cements, microparticles, nanoparticles, and implant coatings. Preclinical and early clinical studies have analyzed phage stability, controlled release, and the synergistic effects of combined phage/antibiotic therapy. However, challenges remain regarding standardization, immunogenicity, and regulatory approval. Nonetheless, phage therapy shows promise for clinical translation as an adjunct or alternative to conventional treatments for OAIs. Well-designed clinical trials are urgently needed to confirm the efficacy of phage therapy, optimize delivery strategies, and integrate the treatments in routine practice. Despite encouraging outcomes for a successful clinical implementation, regulation and standardization of GMP production are required. Full article

Staphylococcus aureus is a major foodborne pathogen associated with contamination of dairy and meat products, posing a persistent challenge to food safety due to its biofilm formation and resistance to multiple antibiotics. In this review, we summarize recent advances in the use of bacteriophages and phage-derived endolysins as targeted biocontrol agents against S. aureus in food systems. Bacteriophages exhibit host specificity and self-replicating capacity, while endolysins provide rapid lytic activity, minimal resistance development, and effectiveness against biofilm-embedded cells. Studies demonstrate significant microbial reductions in milk, cheese, and meat matrices, although factors such as pH, salt, and fat content can influence their efficacy. The integration of these biocontrol tools into food preservation represents a sustainable and safe alternative to conventional antimicrobials. Finally, we discuss current limitations and the need for optimizing formulations, stability, and regulatory frameworks to facilitate the adoption of phage and endolysin-based products in the food industry. Full article

Although phage@nanozymes have proven to be a rapid, precise, and cost-effective method for detecting pathogens in food, the basis of phage and nanozyme selection remains poorly understood. In this study, a novel colourimetric biosensor utilising the temperate phage SapYZUs891 and an Fe/Mn-MOF nanozyme was constructed and assessed for its efficacy in detecting Staphylococcus aureus in food products. Notably, SapYZUs891 exhibited a high titre, broad host range, and strong pH and thermal stability. Moreover, the bimetallic Fe/Mn-MOF nanozyme exhibited an enhanced oxidase-mimicking ability, greater affinity, and a higher reaction rate. The biosensor had a detection time of 19 min, a detection limit of 69 CFU/mL, and a recovery rate between 92.52% and 121.48%, signifying its high reliability and accuracy in identifying S. aureus. This sensor distinguishes between viable and non-viable bacteria and demonstrates resistance to interferent bacterial and food compounds, likely attributable to the particular receptor-binding proteins of SapYZUs891 that bind to the teichoic acid wall on the S. aureus. These results indicated that the SapYZUs891@Fe/Mn-MOF is suitable for the rapid visual assessment of S. aureus. Moreover, the highly sensitive and specific detection system holds significant potential for extended application in on-site screening of S. aureus contamination within food processing environments. Full article

CRISPR-Cas systems are best known as adaptive immune defenses in prokaryotes, but they also function as versatile regulators bridging bacterial immunity with host-related processes. Beyond neutralizing invasive phages and plasmids, these systems influence core aspects of bacterial physiology, such as modulating gene expression, stress responses, biofilm formation, quorum sensing, and virulence. Notably, CRISPR-mediated regulation can facilitate immune evasion at the host-pathogen interface, underscoring these systems as central orchestrators of microbial survival and host interactions. In addition, CRISPR-Cas has rapidly become a cornerstone of synthetic biology and microbiome engineering. Recent strategies repurpose native and engineered CRISPR systems to precisely modulate microbiome composition or deliver sequence-specific antimicrobials, underscoring the expanding translational potential of this system. Collectively, emerging insights highlight both the canonical immune function and non-canonical regulatory roles of CRISPR-Cas, as well as their broad biological and biotechnological relevance. This review provides a critical synthesis of these developments, illustrating how CRISPR-Cas bridges adaptive immunity and microbial physiology, and outlines future directions for harnessing this duality to deepen understanding of microbial physiology and inform new translational applications. Full article

The global surge in multidrug-resistant (MDR) bacterial pathogens has created an urgent imperative for innovative antimicrobial strategies. Enterococcus faecalis, Staphylococcus aureus, and Acinetobacter baumannii demonstrate remarkable antibiotic resistance and dominate hospital-acquired infections. These bacteria often form biofilms, a complex community structure that shields them from immune system phagocytosis, resists antibiotic penetration, and enhances their survival in hostile environments. In clinical cases, these bacteria often form mixed biofilms and lead to treatment failures. Phages and their derivatives have emerged as promising candidates in the fight against drug-resistant bacteria. Lys22, an endolysin derived from an enterococcus phage, has been cloned and demonstrated to possess a broad host range, effectively targeting E. faecalis, various Staphylococcus species, and A. baumannii. When applied to the biofilms formed by these bacteria, Lys22 was found to significantly inhibit both simple and complex biofilms in vitro. Virulent genes, including agrA, sarA, and icaA in S. aureus; asa1, cylA, and gelE in E. faecalis; and OmpA and lpsB in A. baumannii were also downregulated by Lys22. Notably, Lys22 also exhibited a robust protective effect against dual or triple infections involving E. faecalis, S. aureus, and A. baumannii in a zebrafish embryos model, highlighting its potential as a therapeutic agent in combatting multi-bacterial infections. Full article

Preventive immunology is emerging as a cornerstone of animal infectious disease control within One Health, shifting emphasis from treatment to prevention. This review integrates mechanistic insights in host immunity with a comparative evaluation of next-generation interventions—mRNA/DNA and viral-vector vaccines, nanovaccines, monoclonal antibodies, cytokine modulators, probiotics/postbiotics, bacteriophages, and CRISPR-based approaches—highlighting their immunogenicity, thermostability, delivery, and field readiness. Distinct from prior reviews, we appraise diagnostics as preventive tools (point-of-care assays, biosensors, MALDI-TOF MS, AI-enabled analytics) that enable early detection, risk prediction, and targeted interventions, and we map quantifiable links between successful prevention and reduced antimicrobial use. We embed translation factors—regulatory alignment, scalable manufacturing, workforce capacity, equitable access in LMICs, and public trust—alongside environmental and zoonotic interfaces that shape antimicrobial resistance dynamics. We also provide a critical analysis of limitations and failure cases: gene editing may require stacked edits and concurrent vaccination; phage programs must manage host range, resistance, stability, and regulation; and probiotic benefits remain context-specific. Finally, we present a risk–benefit–readiness framework and a time-bound research agenda to guide deployment and evaluation across animal–human–environmental systems. Coordinating scientific innovation with governance and ethics can measurably reduce disease burden, curb antimicrobial consumption, and improve health outcomes across species. Full article

Bacteriophages have been widely investigated as a promising treatment of food, medical equipment, and humans colonized by antibiotic-resistant bacteria. Phages pose particular interest in combating those bacteria which form biofilms, such as the medically important human pathogen Pseudomonas aeruginosa and several plant pathogens, including P. syringae. In an undergraduate lab course, P. putida was used as the host to isolate novel anti-pseudomonal bacteriophages. Environmental samples of soil and water were collected, and purified phage isolates were obtained. After Illumina sequencing, genomes of these phages were assembled de novo and annotated. Assembled genomes were compared with known genomes in the literature and GenBank to identify taxonomic relations and to refine their functional annotations. The thirteen phages described are sipho-, myo-, and podoviruses in several families of Caudoviricetes, spanning several novel genera, with genomes ranging from 40,000 to 96,000 bp. One phage (DDSR119) is unique and is the first reported P. putida siphovirus. The remaining 12 can be clustered into four distinct groups. Six are highly related to each other and to previously described Autotranscriptaviridae phages: Waldo5, PlaquesPlease, and Laces98 all belong to the Waldovirus genus, whereas Stalingrad, Bosely, and Stamos belong to the Troedvirus genus. Zuri was previously classified as the founding member of a new genus Zurivirus within the family Schitoviridae. Ebordelon and Holyagarpour each represent different species within Zurivirus, whereas Meara is a more distantly related member of the Schitoviridae. Dolphis and Jeremy are similar enough to form a genus but have only a few distant relatives among sequenced phages and are notable for being temperate. We identified the lysis cassettes in all 13 phages, compared tail spike structures, and found auxiliary metabolic genes in several. Studies like these, which isolate and characterize infectious virions, enable the identification of novel proteins and molecular systems and also provide the raw materials for further study, evaluation, and manipulation of phage proteins and their hosts. Full article

Lactic acid bacteria (LAB) are central to food fermentation, probiotic delivery, and emerging synthetic biology applications, yet their robust cell envelopes and restriction–modification systems complicate DNA uptake. This review synthesizes practical routes for introducing DNA into LAB—natural competence, electroporation, conjugation, phage-mediated transduction, and biolistics—and outlines vector systems for expression and chromosomal editing, including food-grade strategies. We highlight recent advances that broaden strain tractability while noting strain-to-strain variability and host-specific barriers that still require tailored solutions. These advances directly enable applications in food and probiotic biotechnology, including improving starter robustness, tailoring flavor and texture pathways, and installing food-grade traits without residual selection markers. We close with near-term priorities for standardizing protocols, widening replicon compatibility, and leveraging modern genome-editing platforms to accelerate safe, marker-free engineering of industrial and probiotic LAB. Full article

Bacteriophages, or phages, are sophisticated nanomachines that efficiently infect bacteria. Their infection of lactic acid bacteria (LAB) used in fermentation can lead to significant industrial losses. Among phages that infect monoderm bacteria, those with siphovirion morphology characterized by a long, non-contractile tail are predominant. The initial stage of phage infection involves precise host recognition and binding. To achieve this, phages feature host adhesion devices (HADs) located at the distal end of their tails, which have evolved to recognize specific proteinaceous or saccharidic receptors on the host cell wall. Ceduovirus represents a group of unique lytic siphophages that specifically infect the LAB Lactococcus lactis by targeting proteinaceous receptors. Despite having compact genomes, most of their structural genes are poorly annotated and the architecture and function of their HADs remain unknown. Here we used AlphaFold3 to explore the Ceduovirus HADs and their interaction with the host. We show that Ceduovirus HADs exhibit unprecedented features among bacteriophages infecting Gram⁺, share structural similarities with bacterial secretion system VI, and combine both saccharide and protein-binding modules. Moreover, we could annotate the majority of Ceduovirus genes encoding structural proteins by leveraging their predicted structures, highlighting AlphaFold’s significant contribution to phage genome annotation. Full article

The increasing incidence of hospital-acquired infections and antimicrobial-resistant pathogens poses a major clinical challenge. Nearly all medical devices are vulnerable to bacterial biofilm formation, which acts as a protective coating against the host defense systems and antibiotics. The persistence of biofilm infections, accounting for around 65% of all microbial infections, and poor conventional treatment outcomes has driven interest in alternative approaches like bacteriophage therapy. This review encompasses key aspects of biofilm biology, taking into account the clinically significant ESKAPE pathogens, and provides an in-depth analysis of the role of phage agents in biofilm control as a new biofilm control strategy. Diving deeper into the mechanisms of phage-mediated processes, the review examines how bacteriophages penetrate and disrupt biofilm architecture and evaluates current therapeutic strategies that exploit these actions, acknowledging their limitations and considering possible future directions. Full article

Bacteriophages (phages) play a pivotal role in shaping microbial communities and driving bacterial evolution. Among the diverse mechanisms governing phage–host interactions, the Arbitrium (ARM) communication system represents a recently discovered paradigm in phage decision-making between the lytic and lysogenic cycles. Initially identified in Bacillus-infecting phages, the ARM system employs a quorum-sensing-like peptide signaling mechanism to modulate infection dynamics and optimize population-level survival strategies. Recent studies have elucidated the structural and functional basis of ARM regulation, highlighting its potential applications in antimicrobial therapy, microbiome engineering, and synthetic biology. The significance of ARM systems lies in their ability to regulate bacterial population stability and influence the evolutionary trajectories of microbial ecosystems. Despite being a relatively recent discovery, ARM systems have garnered considerable attention due to their role in decoding phage population dynamics at the molecular level and their promising biotechnological applications. This review synthesizes current advancements in understanding ARM systems, including their molecular mechanisms, ecological implications, and translational potential. By integrating recent findings, we provide a comprehensive framework to guide future research on phage–host communication and its potential for innovative therapeutic strategies. Full article

Escherichia coli O157:H7 has been an important foodborne pathogen causing severe disease in humans worldwide. It is challenging to control E. coli O157:H7 due to its intrinsic acid resistance, ability to survive in various environments, and the emergence of antimicrobial resistance. Recent research showed that phages are promising antibacterial agents. A phage (Φ241) infecting 48 E. coli O157:H7 strains from various sources was previously isolated from an industrial cucumber fermentation at pH 3.7 and 5% NaCl. The efficacy of phage Φ241 infection was evaluated in this study in four representative model food systems (beef broth, cucumber juice, cucumber juice supplemented with NaCl, and apple juice). Pronounced differences in phage effectiveness were found in the tested food systems, and impacted by pH, salinity, and multiplicity of infection. The potential of this phage is evident in beef broth and cucumber juice, where 4- to 6-log reduction in host concentration was achieved within 3 to 5 h. However, apple juice (pH 3.55) completely inhibited host growth and phage infection. Overall, the study shows the high potential of Φ241 as an antibacterial agent to improve food safety. Future research will incorporate a cocktail of phages targeting E. coli O157:H7 to mitigate phage resistance development. Full article

The emergence of multidrug-resistant Salmonella poses a significant threat to global public health and food safety, necessitating the urgent search for new strategies to replace conventional antibiotics. Phages are viruses that can directly target bacteria and have garnered attention in recent years for their development as antibiotic alternatives. In this study, 4458 samples were collected from farms, supermarkets, and human feces, yielding 65 strains of Salmonella, which were serotyped using multiplex PCR. Subsequently, a lytic phage was isolated and identified using the dominant serotype of Salmonella as the host bacterium. We further explored the biological characteristics of this phage through host range, growth properties, and genomic analysis. Finally, we analyzed the potential of the phage to block the cross-host transmission of Salmonella, combining PFGE Salmonella classification, strain sources, and phage lytic phenotypes. The results showed that phage gmqsjt-1 could lyse 69.23% (45/65) of Salmonella, of which 75.56% (34/45) were resistant strains. The optimal multiplicity of infection (MOI) for gmqsjt-1 was 0.01, with a latent period of about 10 min, maintaining high activity within the temperature range of 30 to 60 °C and pH range of 2 to 13. No virulence or resistance genes were detected in the gmqsjt-1 genome, which carries two tail spike proteins (contain FAD binding_2 superfamily, the Tail spike TSP1/Gp66 N-terminal domain, and the Pectin lyase fold) and a holin–lysozyme–spanin lytic system. Phylogenetic classification indicates that phage gmqsjt-1 belongs to a new genus and species of an unnamed family within the class Caudoviricetes. PFGE classification results show a high genetic relationship among human, farm animal, and food source Salmonella, and the comprehensive lytic phenotype reveals that phage gmqsjt-1 can lyse Salmonella with high genetic correlation. These results suggest that this novel lytic Salmonella phage has the potential to inhibit cross-host transmission of Salmonella, making it a promising candidate for developing alternative agents to control Salmonella contamination sources (farms), thereby reducing the risk of human infection with Salmonella through ensuring food system safety. Full article

Gram-negative Burkholderia bacteria are known for causing diseases in humans, animals, and plants, and high intrinsic resistance to antibiotics. Phage therapy is a promising alternative to control multidrug-resistant bacterial pathogens. Here, we present an overview of Burkholderia phage characteristics, host specificity, genomic classification, and therapeutic potentials across medical, veterinary, and agricultural systems. We evaluate the efficacy and limitations of current phage candidates, the biological and environmental barriers of phage applications, and the phage cocktail strategy. We highlight the innovations on the development of targeted phage delivery systems and the transition from the exploration of clinical phage therapy to plant disease management, advocating integrated disease control strategies. Full article