Combination of genetically diverse Pseudomonas phages enhances the cocktail efficiency against bacteria

Scientific Reports, Journal Year: 2023, Volume and Issue: 13(1)

Published: June 1, 2023

Phage treatment has been used as an alternative to antibiotics since the early 1900s. However, bacteria may acquire phage resistance quickly, limiting use of treatment. The combination genetically diverse phages displaying distinct replication machinery in cocktails therefore become a novel strategy improve therapeutic outcomes. Here, we isolated and studied lytic (SPA01 SPA05) that infect wide range clinical Pseudomonas aeruginosa isolates. These relatively small myophages have around 93 kbp genomes with no undesirable genes, 30-min latent period, reproduce high number progenies, ranging from 218 240 PFU per infected cell. Even though both lyse their hosts within 4 h, phage-resistant emerge during Considering SPA01-resistant cross-resist SPA05 vice versa, combining SPA01 for cocktail would be ineffective. According decreased adsorption rate resistant isolates, one anti-phage mechanisms occur through modification receptors on target cells. All however, are susceptible nucleus-forming jumbophages (PhiKZ PhiPA3), which SPA05, suggesting recognize different receptor entry. these jumbophage PhiKZ outperforms other tested combinations terms bactericidal activity effectively suppresses emergence resistance. This finding reveals effectiveness phage-composed reducing bacterial growth prolonging evolution

Language: Английский

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Nucleus-forming jumbophage PhiKZ therapeutically outcompetes non-nucleus-forming jumbophage Callisto

iScience, Journal Year: 2024, Volume and Issue: 27(5), P. 109790 - 109790

Published: April 18, 2024

With the recent resurgence of phage therapy in modern medicine, jumbophages are currently under spotlight due to their numerous advantages as anti-infective agents. However, most significant discoveries date have primarily focused on nucleus-forming jumbophages, not non-nucleus-forming counterparts. In this study, we compare biological characteristics exhibited by two genetically diverse jumbophages: 1) well-studied jumbophage, PhiKZ; and 2) newly discovered Callisto. Single-cell infection studies further show that Callisto possesses different replication machinery, resulting a delay maturation compared PhiKZ. The therapeutic potency both phages was examined

Language: Английский

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Dissecting antibiotic effects on the cell envelope using bacterial cytological profiling: a phenotypic analysis starter kit

Microbiology Spectrum, Journal Year: 2024, Volume and Issue: 12(3)

Published: Jan. 30, 2024

ABSTRACT Phenotypic analysis assays such as bacterial cytological profiling (BCP) have become increasingly popular for antibiotic mode of action analysis. A plethora dyes, protein fusions, and reporter strains are available been used this purpose, enabling both rapid categorization in-depth mechanisms. However, non-expert researchers may struggle choosing suitable interpreting results. This is a particular problem antibiotics that multiple or complex targets, the cell envelope. Here, we set out to curate minimal accessible affordable phenotypic allow distinction between membrane wall can identify dual-action inhibitors, be implemented in most research environments. To end, employed BCP, potential, fluidity, synthesis assays. assess specificity ease interpretation, tested three well-characterized commercially reference antibiotics: potassium ionophore valinomycin, lipid II-binding glycopeptide vancomycin, lantibiotic nisin, which binds II forms pore. Based on our experiments, suggest membrane-potentiometric probe, fluorescent fusions MinD MreB basic assay recommend complementing these with Laurdan-based fluidity measurements PliaI fusion, where indicated. We believe results provide guidance who wish use studies but do not possess specialized equipment expert knowledge employ full breadth possible techniques. IMPORTANCE using fluorescence dyes it difficult implement methods due need and/or complexity biology physiology, making interpretation non-experts. especially problematic compounds pleiotropic effects, inhibitors In order make labs, whose primary expertise biology, limited resources, simple broadly needed easy implement, execute, interpret. curated does highly equipment, performed straightforward interpret without knowing intricacies biology.

Language: Английский

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A novel antibiotic class targeting the enolase of Acinetobacter baumannii

Research Square (Research Square), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 6, 2025

High-throughput screening studies provide an additional approach to discovering repurposed drugs for antimicrobial treatments. In this work, we report the identification of ENOblock, anticancer drug, as a novel antibiotic class. We computationally and experimentally validated that ENOblock synergizes with last resort antibiotic, colistin. Additionally, identified enolase potential bacterial target ENOblock. The in silico vitro antibacterial activity translated into potent vivo efficacy in animal infection models. Collectively, preclinical data support selection promising candidate development, address urgent threat infections caused by Acinetobacter baumannii.

Language: Английский

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Advancing antibiotic discovery with bacterial cytological profiling: a high-throughput solution to antimicrobial resistance

Frontiers in Microbiology, Journal Year: 2025, Volume and Issue: 16

Published: Feb. 13, 2025

Developing new antibiotics poses a significant challenge in the fight against antimicrobial resistance (AMR), critical global health threat responsible for approximately 5 million deaths annually. Finding classes of that are safe, have acceptable pharmacokinetic properties, and appropriately active pathogens is lengthy expensive process. Therefore, high-throughput platforms needed to screen large libraries synthetic natural compounds. In this review, we present bacterial cytological profiling (BCP) as rapid, scalable, cost-effective method identifying antibiotic mechanisms action. Notably, BCP has proven its potential drug discovery, demonstrated by identification cellular target spirohexenolide A methicillin-resistant Staphylococcus aureus . We application different organisms discuss BCP’s advantages, limitations, improvements. Furthermore, highlight studies utilized investigate listed Bacterial Priority Pathogens List 2024 identify whose profiles missing. also explore most recent artificial intelligence deep learning techniques could enhance analysis data generated BCP, potentially advancing our understanding discovery novel druggable pathways.

Language: Английский

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A novel coli myophage and antibiotics synergistically inhibit the growth of the uropathogenic E. coli strain CFT073 in stoichiometric niches

Microbiology Spectrum, Journal Year: 2023, Volume and Issue: 11(5)

Published: Sept. 21, 2023

ABSTRACT Urinary tract infections are widespread bacterial affecting millions of people annually, with Escherichia coli being the most prevalent. Although phage therapy has recently gained interest as a promising alternative for antibiotic-resistant bacteria, several studies have raised concerns regarding evolution resistance, making ineffective. In this study, we discover novel myophage designated Killian that targets E. strains, including uropathogenic (UPEC) strain CFT073. It requires at least 20 minutes 90% its particles to adsorb host cells, undergoes subcellular activities replication 30 minutes, and eventually lyses cells burst size about 139 per cell. Additionally, can withstand wide variety temperatures (4–50°C) pHs ( 4 – 10 ). Genome analysis reveals Killian’s genome consists 169,905 base pairs 35.5% GC content, encoding 276 open reading frames; these, 209 functionally annotated no undesirable genes detected, highlighting potential an antibiotic against UPEC. However, after 8-hour treatment high multiplicities infection, density continuously increases, indicating onset growth revival. Thus, combination study between three different antibiotics, amikacin, ciprofloxacin, piperacillin, was performed showed certain antibiotics exhibited synergistic interactions in suppressing These findings suggest Killian-antibiotic combinations effective inhibiting IMPORTANCE Phage been spotlight viable infections. emergence resistance occurs during treatment, not much effective. Here, present discovery that, by itself, effectively kill , but revival detected treatment. then combined improve efficiency re-growth. This research would pave way future development phage-antibiotic cocktails sustainable use phages therapeutic purposes.

Language: Английский

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Sensing of Antibiotic–Bacteria Interactions

Antibiotics, Journal Year: 2023, Volume and Issue: 12(8), P. 1340 - 1340

Published: Aug. 19, 2023

Sensing of antibiotic–bacteria interactions is an important area research that has gained significant attention in recent years. Antibiotic resistance a major public health concern, and it essential to develop new strategies for detecting monitoring bacterial responses antibiotics order maintain effective antibiotic development antibacterial treatment. This review summarizes advances sensing interactions, which are divided into two main parts: studies on the mechanism action sensitive bacteria interrogation defense mechanisms resistant ones. In conclusion, this provides overview present landscape concerning emphasizing potential method adaptation integration machine learning techniques data analysis, could potentially lead transformative impact mechanistic within field.

Language: Английский

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A marine-derived fatty acid targets the cell membrane of Gram-positive bacteria

Journal of Bacteriology, Journal Year: 2023, Volume and Issue: 205(11)

Published: Oct. 31, 2023

With the lack of new antibiotics in drug discovery pipeline, coupled with accelerated evolution antibiotic resistance, sources that target pathogens clinical importance are paramount. Here, we use bacterial cytological profiling to identify mechanism action monounsaturated fatty acid (

Language: Английский

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Deep Learning-Based Object Detection And Bacteria Morphological Feature Extraction For Antibiotic Mode Of Action Study

Published: Oct. 28, 2023

The increasing prevalence of antimicrobial resistance (AMR), as microorganisms develop to drugs, has emerged a critical concern in infection treatment, resulting rising death toll. Assessing the effect drugs can provide insights by studying morphological change bacteria after drug treatment. However, utilizing conventional techniques such CellProfiler for long-term and large-scale sample experiments is impractical due manual processes involved. To address this challenge, we proposed deep learning-based object detection model predicting type antibiotic treatment automatically extracting morphology. Our combines YOLOX two Cascade R-CNNs using weight box fusion enhance performance. It achieves an mIOU 0.753 mAP 0.699 higher compared (mAP = 0.218). In addition, use computer vision approach extract features including cell membrane, DNA, color intensity classify treated which comparable performance (F1-Score 0.75, 0.79 respectively). We believe our work be used automatic tool efficiency prediction profiles AMR applications. code web application are available at https://github.com/biodatlab/bacteria-detection.

Language: Английский

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Imaging‐based profiling for elucidation of antibacterial mechanisms of action

Biotechnology and Applied Biochemistry, Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 28, 2024

Abstract In this review, we aim to summarize experimental data and approaches identifying cellular targets or mechanisms of action antibacterials based on imaging techniques. Imaging‐based profiling methods, such as bacterial cytological profiling, dynamic morphology imaging, others, have become a useful research tool for mechanistic studies new antibiotics well combinations with conventional ones other therapeutic options. The main methodological details obtained results are summarized discussed. review covers the literature up February 2024.

Language: Английский

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