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Keywords = bacteriophage isolation

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33 pages, 2923 KB  
Article
Co-Occurrence of Toxic Bloom-Forming Cyanobacteria Planktothrix, Cyanophage, and Symbiotic Bacteria in Ohio Water Treatment Waste: Implications for Harmful Algal Bloom Management
by Angela Brooke Davis, Morgan Evans, Katelyn McKindles and Jiyoung Lee
Toxins 2025, 17(9), 450; https://doi.org/10.3390/toxins17090450 - 5 Sep 2025
Abstract
Cyanobacterial blooms are increasingly becoming more intense and frequent, posing a public health threat globally. Drinking water treatment plants that rely on algal bloom-affected waters may create waste (water treatment residuals, WTRs) that concentrates contaminants. Source waters may contain harmful cyanobacteria, cyanophages (bacteriophages [...] Read more.
Cyanobacterial blooms are increasingly becoming more intense and frequent, posing a public health threat globally. Drinking water treatment plants that rely on algal bloom-affected waters may create waste (water treatment residuals, WTRs) that concentrates contaminants. Source waters may contain harmful cyanobacteria, cyanophages (bacteriophages that infect cyanobacteria), and bacteria. Cyanophages are known to affect bloom formation and growth dynamics, so there is a need to understand viral-host dynamics between phage and bacteria in these ecosystems for managing cyanobacteria. This study isolated and characterized lytic cyanophages from WTRs of a HAB-affected lake in Ohio that infect toxic bloom-forming filamentous cyanobacteria Planktothrix agardhii. Phage infections in the Lake Erie cyanobacteria culture were examined visually and via microscopy and fluorometry. Whole genome sequencing and metagenomic analyses were also conducted. Observed changes in Planktothrix included sheared and shriveled filaments, reduced clumping, and buoyancy changes. Photosynthetic pigmentation was unexpectedly more apparent during phage infection. Metagenomic analyses identified nineteen phages and seven other co-existing bacterial genera. Annotated bacterial genomes contained metabolic pathways that may influence phage infection efficiency. Viral genomes were successfully tied to microbial hosts, and annotations identified important viral infection proteins. This study examines cyanobacterial-phage interactions that may have potential for bioremedial applications. Full article
30 pages, 16346 KB  
Article
Isolation of Lytic Bacteriophages of Escherichia coli and Their Combined Use with Antibiotics Against the Causative Agents of Colibacillosis in Calves
by Pavel G. Alexyuk, Andrey P. Bogoyavlenskiy, Kuralay S. Akanova, Yergali S. Moldakhanov, Timur T. Kerimov, Nadezhda S. Sokolova, Vladimir E. Berezin and Madina S. Alexyuk
Vet. Sci. 2025, 12(9), 817; https://doi.org/10.3390/vetsci12090817 - 26 Aug 2025
Viewed by 416
Abstract
The spread of antibiotic-resistant pathogenic Escherichia coli poses a serious threat to calf health on livestock farms. With the decline in antibiotic therapy effectiveness, alternative approaches such as phage therapy are urgently needed. This study aimed to isolate lytic E. coli bacteriophages, characterize [...] Read more.
The spread of antibiotic-resistant pathogenic Escherichia coli poses a serious threat to calf health on livestock farms. With the decline in antibiotic therapy effectiveness, alternative approaches such as phage therapy are urgently needed. This study aimed to isolate lytic E. coli bacteriophages, characterize their properties, and evaluate the synergistic effects of their combined use with veterinary antibiotics against colibacillosis pathogens in calves. As a result of the work, 4 bacteriophages were isolated from wastewater from various cities of Kazakhstan: vB_EcoS_ABO/4, vB_EcoM_PL/4, vB_Eco_CWW/26, vB_EcoM_ShWW/46. Morphological, biological, and genomic analyses showed that the phages belong to different genera of the Caudoviricetes class, possess high lytic activity, broad host range, environmental stability, and lack genes associated with lysogeny, antibiotic resistance, or virulence. Interaction studies with antibiotics revealed synergistic or additive effects in over 75% of cases. These findings highlight the strong potential of the isolated bacteriophages for independent or adjunctive use in the treatment and prevention of colibacillosis in calves. However, further in vivo studies are required to definitively confirm their therapeutic efficacy. Full article
(This article belongs to the Section Veterinary Biomedical Sciences)
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15 pages, 2298 KB  
Article
Unveiling the Petunia hybrida Virome: Metatranscriptomic Profiling from the Bulgarian Market and In Vitro Cultures
by Rumyana Valkova, Stoyanka Jurak, Elena Apostolova-Kuzova, Vesselin Baev, Lilyana Nacheva, Galina Yahubyan, Dijana Škorić and Mariyana Gozmanova
Plants 2025, 14(16), 2597; https://doi.org/10.3390/plants14162597 - 21 Aug 2025
Viewed by 349
Abstract
RNA sequencing is a high-throughput sequencing method essential for unbiased detection and characterization of known and emerging plant viruses. Its high sensitivity makes it particularly well-suited for identifying low-abundance viral sequences, even in asymptomatic plants or those affected by complex, mixed infections. Here, [...] Read more.
RNA sequencing is a high-throughput sequencing method essential for unbiased detection and characterization of known and emerging plant viruses. Its high sensitivity makes it particularly well-suited for identifying low-abundance viral sequences, even in asymptomatic plants or those affected by complex, mixed infections. Here, we conducted a metatranscriptomic survey of Petunia hybrida plants from the Bulgarian market, both symptomatic and asymptomatic, and their corresponding in vitro plantlets. Viruses were detected in all tested samples demonstrating that visual symptoms are not a reliable indicator of infection. The viromes were dominated by petunia vein clearing virus (PVCV, Petuvirus venapetuniae), cucumber mosaic virus (CMV, Cucumovirus CMV), and tomato aspermy virus (TAV, Cucumovirus TAV), along with bacteriophages and fungus-associated viruses. However, the PVCV and CMV abundance was elevated in in vitro samples, possibly due to cutting-induced activation and/or prolonged cultivation. Phylogenetic analysis of the Bulgarian CMV, TAV, and PVCV isolates highlights their genetic links to strains from a wide geographic range and diverse hosts, emphasizing the potential for virus movement and genetic exchange among plant viruses across regions and species. It also suggests that petunias may contribute to the transmission dynamics of viruses within ornamental trade networks. These findings also emphasize the phytosanitary risks to horticulture and establish a basis for further investigation into plant virus ecology. Full article
(This article belongs to the Special Issue Virus-Induced Diseases in Horticultural Plants)
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10 pages, 1209 KB  
Article
Feasibility and Potential Clinical Ramifications of Using Bacteriophage Therapy for S. aureus Necrotizing Fasciitis
by James B. Doub, Dakarai Dunbar, Sara Jain, Maggie Manchester, Lila Berle, Janvi Madhiwala, Bradley Anderson, Riva Malick, Max Jacobs and Kenneth L. Urish
J. Clin. Med. 2025, 14(16), 5609; https://doi.org/10.3390/jcm14165609 - 8 Aug 2025
Viewed by 367
Abstract
Background: Staphylococcus aureus necrotizing fasciitis is a life-threatening infection requiring aggressive surgical and medical management. Despite these interventions, tremendous morbidity and mortality occur. Thus, novel agents are needed to reduce these negative outcomes. Consequently, the aims of this translational study were to [...] Read more.
Background: Staphylococcus aureus necrotizing fasciitis is a life-threatening infection requiring aggressive surgical and medical management. Despite these interventions, tremendous morbidity and mortality occur. Thus, novel agents are needed to reduce these negative outcomes. Consequently, the aims of this translational study were to evaluate the feasibility of using bacteriophages and the potential clinical ramifications of using bacteriophages in treatment of S. aureus necrotizing fasciitis. Methods: Necrotizing fasciitis clinical isolates (n = 6) were tested against different Staphylococcal bacteriophages (n = 4) to assess for activity. After exposure to bacteriophages that had growth inhibition for more than 16 h, the ability of S. aureus to change phenotypic expression of numerous enzymes was evaluated, and the ability to reduce bacterial virulence was measured with the Caenorhabditis elegans assay. Results: Staphylococcal myoviridae bacteriophages were able to lyse most clinical isolates (83%). Interestingly, after exposure to myoviridae bacteriophages, S. aureus isolates had no expression of hemolysin, secreted coagulase, or lecthinase, or the ability to ferment mannitol. These same bacteriophages also caused statistically significant decreases in bacterial virulence (p < 0.05). Neither findings were observed for bacteriophages of the podoviridae family. Conclusions: To use bacteriophages for S. aureus necrotizing fasciitis, cocktails of Staphylococcal myoviridae are likely needed to allow for broad host ranges, mitigating the need for in vitro sensitivity testing. Moreover, Staphylococcal myoviridae have the potential to reduce specific enzyme expression and global virulence of residual S. aureus. Thus, bacteriophages may aid in reducing necrotizing fasciitis morbidity by not only lysing S. aureus but also by reducing S. aureus virulence. Full article
(This article belongs to the Special Issue Clinical Advances in Orthopedic Infections)
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24 pages, 10760 KB  
Article
Pseudomonas Phage Banzai: Genomic and Functional Analysis of Novel Pbunavirus with Lytic Activity Against Pseudomonas aeruginosa
by Andrei V. Chaplin, Nina N. Sykilinda, George A. Skvortsov, Konstantin S. Troshin, Anna A. Vasilyeva, Sofia A. Shuraleva, Artem A. Malkov, Vladislav S. Simonov, Boris A. Efimov, Lyudmila I. Kafarskaia, Konstantin A. Miroshnikov, Anna A. Kuznetsova and Peter V. Evseev
Viruses 2025, 17(8), 1088; https://doi.org/10.3390/v17081088 - 6 Aug 2025
Viewed by 705
Abstract
Antibiotic-resistant Pseudomonas aeruginosa presents a critical global health challenge, particularly in hospital-acquired infections. Bacteriophages offer a promising therapeutic avenue due to their ability to target and lyse resistant strains. This study characterizes Pseudomonas phage Banzai, a newly isolated Pbunavirus (family Lindbergviridae) with [...] Read more.
Antibiotic-resistant Pseudomonas aeruginosa presents a critical global health challenge, particularly in hospital-acquired infections. Bacteriophages offer a promising therapeutic avenue due to their ability to target and lyse resistant strains. This study characterizes Pseudomonas phage Banzai, a newly isolated Pbunavirus (family Lindbergviridae) with lytic activity against multiple P. aeruginosa isolates, including multidrug-resistant strains. Genomic analysis revealed a 66,189 bp genome, lacking antibiotic resistance or virulence factors, and suggested a headful packaging mechanism and the presence of a bidirectional component in the replication. In vivo experiments using Galleria mellonella showed therapeutic potential, significantly improving larval survival (87% at 24 h). Host range analysis revealed activity against 13 of 30 P. aeruginosa isolates, including members of O1, O3, O5 and O6 in silico predicted serogroups. Phylogenomic analyses place phage Banzai within the genus Pbunavirus, sharing 94.8% intergenomic similarity with its closest relatives, supporting its classification as a novel species. These findings highlight phage Banzai as a potential candidate for phage therapy, demonstrating genomic stability, a strictly lytic lifestyle, and in vivo efficacy. Full article
(This article belongs to the Section Bacterial Viruses)
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16 pages, 2146 KB  
Article
Isolation and Characterization of a Cold-Adapted Bacteriophage for Biocontrol of Vibrio parahaemolyticus in Seafood
by Zhixiang Nie, Xiangyu Cheng, Shengshi Jiang, Zhibin Zhang, Diwei Zhang, Hanfang Chen, Na Ling and Yingwang Ye
Foods 2025, 14(15), 2660; https://doi.org/10.3390/foods14152660 - 29 Jul 2025
Viewed by 479
Abstract
Vibrio parahaemolyticus (V. parahaemolyticus) is a preeminent seafood-borne pathogen, imposing significant economic burdens on global aquaculture. The escalating prevalence of multidrug-resistant strains has accentuated the critical urgency for developing sustainable biocontrol strategies. In this study, a bacteriophage designated vB_VPAP_XY75 (XY75) was [...] Read more.
Vibrio parahaemolyticus (V. parahaemolyticus) is a preeminent seafood-borne pathogen, imposing significant economic burdens on global aquaculture. The escalating prevalence of multidrug-resistant strains has accentuated the critical urgency for developing sustainable biocontrol strategies. In this study, a bacteriophage designated vB_VPAP_XY75 (XY75) was isolated and biologically characterized to establish an effective control against V. parahaemolyticus. XY75 exhibited remarkable specificity toward V. parahaemolyticus, effectively lysing 46.2% of the target strains while showing no lytic activity against non-target bacterial species. Morphological characterization confirmed its taxonomic assignment to the Myoviridae family, featuring an icosahedral head (40 ± 2 nm) and contractile tail (60 ± 2 nm). XY75 demonstrated strong environmental tolerance, remaining stable at pH 4–11 and temperatures as high as 50 °C. At an optimal multiplicity of infection (MOI = 0.01), XY75 achieved a peak titer of 8.1 × 1010 PFU/mL, a 5 min latent period, and burst size of 118 PFU/cell. Critically, XY75 reduced V. parahaemolyticus in salmon by more than 5.98 log CFU/g (99.9%) within 6 h at 4 °C, demonstrating exceptional cold tolerance and lytic activity. Genomic analysis confirmed that no virulence or antibiotic resistance genes were present. These results establish XY75 as a safe and efficacious biocontrol candidate for seafood preservation, with particular utility under refrigerated storage conditions. Full article
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18 pages, 682 KB  
Article
Antimicrobial Potential of Bacteriophages JG005 and JG024 Against Pseudomonas aeruginosa Isolates from Canine Otitis
by Maura R. Lourenço, Eva Cunha, Luís Tavares and Manuela Oliveira
Vet. Sci. 2025, 12(7), 646; https://doi.org/10.3390/vetsci12070646 - 7 Jul 2025
Viewed by 1114
Abstract
Canine otitis externa caused by Pseudomonas aeruginosa is a relevant disease in veterinary medicine. Given P. aeruginosa’s high priority status for the development of new antimicrobials, innovative strategies like bacteriophage therapy are essential. Lytic bacteriophages are viruses with high specificity for their bacterial [...] Read more.
Canine otitis externa caused by Pseudomonas aeruginosa is a relevant disease in veterinary medicine. Given P. aeruginosa’s high priority status for the development of new antimicrobials, innovative strategies like bacteriophage therapy are essential. Lytic bacteriophages are viruses with high specificity for their bacterial hosts, making them a promising therapeutic choice in both human and veterinary medicine. This study aimed to evaluate the antimicrobial potential of bacteriophages JG005 and JG024, first characterized in terms of their biofilm-forming ability and antimicrobial susceptibility profile, against P. aeruginosa isolates obtained from dogs with otitis externa,. Bacteriophages titer, host range, and activity were assessed against P. aeruginosa biofilms via microtiter assays using crystal violet and Alamar Blue. JG024 showed lytic activity against 61.2% (n = 30/49) of the isolates, while JG005 showed lytic activity against 38.8% (n = 19/49) of the isolates. Crystal violet quantification showed that JG005 can promote strong microbial suppression of 60% (n = 6/10) and 50% (n = 5/10) of the isolates at a multiplicity of infection (MOI) of 10 and 100, respectively. JG024 presented strong microbial suppression of 20% (n = 2/10) of the isolates regardless of the MOI level tested. These phages show promising potential as an innovative treatment for canine otitis externa caused by P. aeruginosa, but further studies are needed before future clinical use. Full article
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16 pages, 1769 KB  
Article
Isolation and Characterization of a Crude Oil-Tolerant Obligate Halophilic Bacterium from the Great Salt Lake of the United States of America
by Jonathan Oakes, Johurimam Noah Kuddus, Easton Downs, Clark Oakey, Kristina Davis, Laith Mohammad, Kiara Whitely, Carl E. Hjelmen and Ruhul Kuddus
Microorganisms 2025, 13(7), 1568; https://doi.org/10.3390/microorganisms13071568 - 3 Jul 2025
Viewed by 521
Abstract
Most large-scale crude oil spills occur in marine environments. We screened easily propagable/maintainable halophiles to develop agents for the bioremediation of marine spills. A bacterial strain isolated from a polluted region of the Great Salt Lake was characterized and tested for its ability [...] Read more.
Most large-scale crude oil spills occur in marine environments. We screened easily propagable/maintainable halophiles to develop agents for the bioremediation of marine spills. A bacterial strain isolated from a polluted region of the Great Salt Lake was characterized and tested for its ability to degrade crude oil. The strain (Salinivibrio costicola) is motile, catalase- and lipase-positive, a facultative anaerobe, and an obligate halophile. Its growth optimum and tolerance ranges are: NaCl (5%, 1.25–10%), pH (8, 6–10), and temperature (22 °C, 4–45 °C). Its genome (3,166,267 bp) consists of two circular chromosomes and a plasmid, containing 3197 genes, including some genes potentially relevant to hydrocarbon metabolism. The strain forms a biofilm but is considered nonpathogenic and is sensitive to some common antibiotics. Lytic bacteriophages infecting the strain are rare in the water samples we tested. The strain survived on desiccated agar media at room temperature for a year, grew optimally in complex media containing 0.1–1% crude oil, but failed to reduce total recoverable petroleum hydrocarbons from crude oil. Thus, a recalcitrant halophile may endure crude oil without mineralizing. Due to some of their advantageous attributes, such strains can be considered for genetic manipulation to develop improved agents for bioremediation. Full article
(This article belongs to the Special Issue Marine Microbes, Biocontamination and Bioremediation)
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16 pages, 3289 KB  
Article
Transcriptomic Analysis of Biofilm Formation Inhibition by PDIA Iminosugar in Staphylococcus aureus
by Anna Tomusiak-Plebanek, Łucja Kozień, Estelle Gallienne, Maciej Florczyk, Sławomir Ciesielski, Piotr Heczko and Magdalena Strus
Antibiotics 2025, 14(7), 668; https://doi.org/10.3390/antibiotics14070668 - 1 Jul 2025
Viewed by 457
Abstract
Background: Iminosugars are natural or synthetic sugar analogues with a very broad spectrum of activities, including those against the most prominent bacterial pathogens, like P. aeruginosa or S. aureus. In a series of studies, we have demonstrated that one of the synthetic iminosugars, [...] Read more.
Background: Iminosugars are natural or synthetic sugar analogues with a very broad spectrum of activities, including those against the most prominent bacterial pathogens, like P. aeruginosa or S. aureus. In a series of studies, we have demonstrated that one of the synthetic iminosugars, PDIA (beta-1-C-propyl-1,4-dideoxy-1,4-imino-L-arabinitol), possesses the ability to suppress biofilm production by different pathogenic bacteria without inhibiting their growth. Thereby, PDIA is able to influence experimental skin infection caused by S. aureus. Methods: To elucidate molecular mechanisms by which PDIA impedes biofilm formation by S. aureus, a transcriptomic study was performed in which a biofilm-producing S. aureus strain was grown in the presence of PDIA for 24 and 48 h in comparison to a control without the iminosugar. The RNA was then isolated, converted into cDNA, sequenced, and data analysis was performed. Results: It appeared that PDIA caused the down-regulation of many bacteriophage genes responsible for the processes of bacterial cell lysis, and some genes responsible for cell wall degradation were also down-regulated. Among the 25 most upregulated genes were those representing the phosphotransferase system (PTS), which is required for carbohydrate uptake and control of carbon metabolism. The ranking of the most significant down-regulated genes after 24 h exposure to PDIA shows that they predominantly coded for both the synthesis and lysis of the peptidoglycan. Conclusions: We have shown here that the influence of PDIA on the expression of S. aureus genes is broad and affects many genes encoding metabolism and ribosomes. Full article
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21 pages, 4687 KB  
Article
The Ability of Bacteriophages to Reduce Biofilms Produced by Pseudomonas aeruginosa Isolated from Corneal Infections
by Kuma Diriba Urgeya, Dinesh Subedi, Naresh Kumar and Mark Willcox
Antibiotics 2025, 14(7), 629; https://doi.org/10.3390/antibiotics14070629 - 20 Jun 2025
Viewed by 1635
Abstract
Pseudomonas aeruginosa (P. aeruginosa) is a common antibiotic-resistant pathogen, posing significant public health threats worldwide. It is a major cause of ocular infections, mostly linked to contact lens wear. P. aeruginosa often produces biofilm during infections, and these are also associated [...] Read more.
Pseudomonas aeruginosa (P. aeruginosa) is a common antibiotic-resistant pathogen, posing significant public health threats worldwide. It is a major cause of ocular infections, mostly linked to contact lens wear. P. aeruginosa often produces biofilm during infections, and these are also associated with antibiotic resistance. Bacteriophage (phage) therapy is emerging as a promising approach for treating multidrug-resistant P. aeruginosa. Objective: This study aimed to assess the antibiofilm effects of six phages against P. aeruginosa biofilms isolated from patients with corneal infections. Method: This study examined P. aeruginosa strains for their ability to form biofilms using crystal violet assay. Six P. aeruginosa bacteriophages (DiSu1 to DiSu6) were used, which were isolated from sewage water in Melbourne, Australia. Spot tests were used to assess phage sensitivity. The effect of phages against P. aeruginosa strains was determined using time–kill assay and efficiency of plating. The ability of phage to inhibit biofilm formation over 24 h or reduce preformed biofilms was also studied and confirmed using confocal laser scanning microscopy with Live/Dead staining. Result: After 24 h of incubation, all tested P. aeruginosa strains formed moderate to strong biofilms. All P. aeruginosa strains were sensitive to at least four of the six phages. The highest level of bacterial growth inhibition in the liquid infection model was observed when phages were applied at a multiplicity of infection (MOI) of 100. Certain bacteria/phage combinations were able to inhibit biofilm formation over 24 h, with the combination of strain PA235 and phage DiSu3 producing the highest inhibition (83%) at a MOI of 100. This was followed by the combinations of PA223/DiSu3 (56%), and PA225/DiSu5 (52%). For the reduction in preformed biofilms, the best combinations were PA235 (90%), PA221 (61%), and PA213 and PA225 (57% each), all with DiSu3 after 3 h. However, exposing the biofilm with phages for over 24 h appeared to promote phage resistance as there was evidence of biofilm growth, with the only combination still showing a significant reduction being PA221/DiSu3 (58%) at MOI of 100. Conclusions: This study showed that the effect of phages against P. aeruginosa is concentration (MOI) dependent. Phages at higher MOI have the ability to disrupt, inhibit, and reduce P. aeruginosa biofilms. However, prolonged exposure of the biofilm with phages appeared to promote phage resistance. To enhance phage efficacy and address this form of resistance, further studies utilizing phage cocktails or a combination of phages and antibiotics is warranted. Full article
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16 pages, 3913 KB  
Article
Isolation and Characterization of Enterococcus faecalis Phage ZXL-01 and Preliminary Investigation of Its Therapeutic Effect on Periapical Periodontitis
by Hailin Jiang, Xueli Zhao, Chuhan Wang, Hongyan Shi, Jinghua Li, Chunyan Zhao and Honglan Huang
Curr. Issues Mol. Biol. 2025, 47(6), 469; https://doi.org/10.3390/cimb47060469 - 18 Jun 2025
Viewed by 516
Abstract
Enterococcus faecalis (E. faecalis) is a major pathogen responsible for refractory apical periodontitis (RAP). It can penetrate deep into dentinal tubules, form persistent biofilms, and exhibit antibiotic resistance, thereby limiting the efficacy of conventional antimicrobial treatments. Bacteriophages (phages), due to their [...] Read more.
Enterococcus faecalis (E. faecalis) is a major pathogen responsible for refractory apical periodontitis (RAP). It can penetrate deep into dentinal tubules, form persistent biofilms, and exhibit antibiotic resistance, thereby limiting the efficacy of conventional antimicrobial treatments. Bacteriophages (phages), due to their strong lytic activity and host specificity, have emerged as promising alternatives. In this study, a novel strictly lytic phage, ZXL-01, was isolated from lake water in Jilin, China. ZXL-01 demonstrated remarkable stability under extreme conditions, including thermal tolerance at 60 °C for 1 h and a wide pH range (4–11). Whole-genome sequencing (GenBank accession number: ON113334) revealed a genome of 40,804 bp with no virulence or tRNA genes, confirming its identity as an E. faecalis phage. Importantly, ZXL-01 exhibited potent antibiofilm activity, reducing biofilm biomass by approximately 69.4% in the inhibition group and 68.4% in the lysis group (both p < 0.001). In an in vitro root canal infection model induced by E. faecalis, scanning electron microscope (SEM) observations confirmed that ZXL-01 effectively inhibited biofilm formation and disrupted mature biofilms. These findings highlight the potential of ZXL-01 as a novel antimicrobial agent for the treatment of E. faecalis-associated apical periodontitis. Full article
(This article belongs to the Section Molecular Microbiology)
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12 pages, 2531 KB  
Article
Isolation and Characterization of Aeromonas salmonicida Phage TSW001 and Its Application on Large Yellow Croaker
by Jun Yan, Zhenghao Guo and Jing Xie
Foods 2025, 14(12), 2082; https://doi.org/10.3390/foods14122082 - 12 Jun 2025
Viewed by 681
Abstract
Aeromonas salmonicida is a common spoilage bacterium found in refrigerated fish. In this study, a virulent bacteriophage was isolated from wastewater using A. salmonicida AS08 as the host, and it was designated as TSW001. Based on morphological characterization and whole-genome analysis, bacteriophage TSW001 [...] Read more.
Aeromonas salmonicida is a common spoilage bacterium found in refrigerated fish. In this study, a virulent bacteriophage was isolated from wastewater using A. salmonicida AS08 as the host, and it was designated as TSW001. Based on morphological characterization and whole-genome analysis, bacteriophage TSW001 was classified within the genus Tedavirus. Biological characterization revealed that TSW001 maintained a stable titer within a temperature range of 4~60 °C, a pH range of 4~9, and a salinity range of 50~1000 mM. The optimal multiplicity of infection (MOI) for TSW001 was 0.1, with a short latency period of approximately 10 min and a burst size of approximately 68 PFU/cell. When applied during the cold storage of large yellow croaker, the A. salmonicida count in the fish juice decreased by approximately 2.1~2.3 log10 CFU/mL over the first two days, while the count in the fish fillets decreased by approximately 1.1~1.8 log10 CFU/g. Furthermore, TSW001 demonstrated the ability to inhibit the formation of A. salmonicida biofilms. These results suggest that phage TSW001 is a promising biological antimicrobial agent for controlling A. salmonicida during the cold storage of seafood. Full article
(This article belongs to the Section Food Quality and Safety)
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18 pages, 1505 KB  
Review
Bacteriophages as Potential Anti-Pathogenic Agents for Intestinal Health of Weaned Piglets in the Post-Antibiotic Era: An Updated Review
by Jun Chen, Jiajun Han, Zheng Yang, Wenyue Zhou, Yuyong He, Xingping Chen, Xin Li, Tiande Zou and Jinming You
Animals 2025, 15(12), 1713; https://doi.org/10.3390/ani15121713 - 10 Jun 2025
Viewed by 602
Abstract
Background: The use of in-feed antibiotics was banned in numerous countries within the animal production industry as a result of the emergence of antibiotic-resistant bacteria and the presence of residual antibiotics. Bacteriophages, which are viruses that infect host bacterial cells, are considered the [...] Read more.
Background: The use of in-feed antibiotics was banned in numerous countries within the animal production industry as a result of the emergence of antibiotic-resistant bacteria and the presence of residual antibiotics. Bacteriophages, which are viruses that infect host bacterial cells, are considered the natural predators of bacteria. Over the past two decades, bacteriophages have garnered increasing attention for their potential in controlling pathogenic bacteria in weaned piglets. Aims: The aim of this overview was to update the progress of bacteriophage application in weaned piglets. Methods: For the section on bacteriophage application in weaned piglets, a systematic search was performed to identify relevant articles published before June 2025 in databases such as Web of Science. Results: In this review, we provide a brief overview of bacteriophages, followed by a summary of the isolation of specific bacteriophages in weaned piglets. In addition, we have summarized the application progress of bacteriophages in weaned piglets, including the effects of oral administration or dietary supplementation with bacteriophages on growth performance, diarrhea characteristics, intestinal morphology, intestinal pH, nutrient digestibility, inflammatory response, intestinal barrier function, and intestinal microecology. Conclusions: This updated overview novelly highlights the potential of bacteriophages as anti-pathogenic agents in mitigating infections caused by pathogenic bacteria in weaned piglets. This review could provide a scientific basis for controlling pathogenic bacteria infections in weaned piglets in the post-antibiotic era. Full article
(This article belongs to the Topic Feeding Livestock for Health Improvement)
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16 pages, 9902 KB  
Article
Genome Sequences of the First Phages Infecting Limnohabitans Reveal Their Global Distribution and Metabolic Potential
by Boxuan Deng, Raoqiong Che, Pinxin Zhu, Yongxia Wang, Zhiying Li, Shiying Zhang and Wei Xiao
Microorganisms 2025, 13(6), 1324; https://doi.org/10.3390/microorganisms13061324 - 6 Jun 2025
Viewed by 664
Abstract
Bacteriophages (phages) are one of the critical biotic drivers of prokaryotic community dynamics, functions, and evolution. Despite their importance in aquatic ecosystems, very few phages have been isolated from freshwater lakes, hampering our understanding of their ecological importance and usage in a variety [...] Read more.
Bacteriophages (phages) are one of the critical biotic drivers of prokaryotic community dynamics, functions, and evolution. Despite their importance in aquatic ecosystems, very few phages have been isolated from freshwater lakes, hampering our understanding of their ecological importance and usage in a variety of biotechnological applications. Limnohabitans, with a ubiquitous distribution, is a metabolically versatile, fast-growing, morphologically diverse freshwater lake bacterial genera. It is especially abundant in pH-neutral and alkaline aquatic habitats, where it represents an average of 12% of freshwater bacterioplankton and plays an important role in funneling carbon from primary producers to higher trophic levels. However, no phages infecting Limnohabitans have been reported to date. Here, we describe, for the first time, three phages infecting Limnohabitans, DC31, DC33, and YIMV22061, isolated from two freshwater lakes in China and characterized using genome content analysis and comparative genomics. DC31 and DC33, recovered from the eutrophic Dianchi Lake, with auxiliary metabolic genes (AMGs), associated with nucleotide metabolism, whereas YIMV22061, isolated from the oligotrophic Fuxian Lake, carried AMGs involved in antibiotic resistance. The AMGs they carried highlight their impacts on Limnohabitans in different environments. Comparative genomic analyses indicate that DC31, DC33, and YIMV22061 represent three novel species in the Caudoviricetes class. IMG/VR database alignment further reveal that these phages are widely distributed across diverse aquatic and terrestrial ecosystems globally, suggesting their ecological significance. This study provides a basis for better understanding Limnohabitans–phage interactions. Full article
(This article belongs to the Special Issue Advances in Genomics and Ecology of Environmental Microorganisms)
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18 pages, 2973 KB  
Article
A TAT Peptide-Functionalized Liposome Delivery Phage System (TAT-Lip@PHM) for an Enhanced Eradication of Intracellular MRSA
by Kaixin Liu, Xin Lu, Xudong Guo, Yi Yang, Wanying Liu, Hongbin Song and Rongtao Zhao
Pharmaceutics 2025, 17(6), 743; https://doi.org/10.3390/pharmaceutics17060743 - 5 Jun 2025
Viewed by 646
Abstract
Background: Intracellular bacteria frequently result in chronic and recurrent infections. MRSA is one of the most prevalent facultative intracellular bacteria in clinical infections. The drug resistance of MRSA and the difficulty of most antibiotics in entering cells result in a suboptimal clinical efficacy [...] Read more.
Background: Intracellular bacteria frequently result in chronic and recurrent infections. MRSA is one of the most prevalent facultative intracellular bacteria in clinical infections. The drug resistance of MRSA and the difficulty of most antibiotics in entering cells result in a suboptimal clinical efficacy of antibiotics in the treatment of intracellular MRSA. Bacteriophages represent a promising alternative therapy in the context of the current antimicrobial resistance crisis. Nevertheless, the low efficiency of phage entry into cells and their rapid inactivation remain challenges in the treatment of intracellular MRSA using phages. The utilization of functionalized carriers for the delivery of phages into cells and their protection represents a feasible strategy. Methods: In this study, a new MRSA bacteriophage (vB_SauS_PHM) was isolated from hospital sewage, exhibiting the characteristics of short incubation period, large lytic amount, and good environmental tolerance. Subsequently, vB_SauS_PHM was encapsulated by TAT peptide-functionalized liposomes through microfluidic technology and size-exclusion chromatography (SEC), forming a phage delivery system, designated TAT-Lip@PHM. Results: The encapsulation rate of the phage by TAT-Lip@PHM was 20.3%, and the cell entry efficiency was ≥90% after 8 h. The 24 h eradication rate of 300 μg/mL TAT-Lip@PHM against intracellular MRSA was 94.05% (superior to the 21.24% and 44.90% of vB_SauS_PHM and Lip@PHM, respectively), while the mammalian cell activity was >85% after 24 h incubation. Conclusions: The TAT-Lip@PHM effectively delivered the phage into the cell and showed an excellent killing effect on intracellular MRSA with low cytotoxicity. This work provides a technical reference for the application of phages in the treatment of intracellular bacterial infection. Full article
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