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Su J, Tan Y, Liu S, Zou H, Huang X, Chen S, Zhang H, Li S, Zeng H. Characterization of a novel lytic phage vB_AbaM_AB4P2 encoding depolymerase and its application in eliminating biofilms formed by Acinetobacter baumannii. BMC Microbiol 2025; 25:123. [PMID: 40057696 PMCID: PMC11889872 DOI: 10.1186/s12866-025-03854-3] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/16/2024] [Accepted: 02/28/2025] [Indexed: 05/13/2025] Open
Abstract
BACKGROUND Acinetobacter baumannii strains are a primary cause of hospital-acquired infections. This bacterium frequently causes biofilm-related infections, notably ventilator-associated pneumonia and catheter-related infections, which exhibit remarkable resistance to antibiotic treatment, posing a severe challenge in the prevention of A. baumannii infections. Therefore, strategies to eliminate the biofilm of A. baumannii in catheters are becoming increasingly important. Phages are capable of lysing bacteria and have a certain effect on the ablation of biofilms. METHODS Sewage treatment plant water was collected for the isolation of A. baumannii phages. The morphological, host range, one-step growth, temperature and pH stability, bactericidal activity, sequencing and genomic analysis were performed to characterize the isolated phage. The three-dimensional structure of the tail fiber protein was predicted by AlphaFold3. The efficacy of phage in clearing biofilms of A. baumannii from 24-well plates and PVC catheters was also evaluated. RESULTS In this study, A. baumannii lytic phage vB_AbaM_AB4P2 was isolated from sewage treatment plant water, showing a clear plaque with halo zone. One-step growth assays unveiled a 20-minute latent period and a burst size of 61 plaque forming unit/cell (PFU/cell). At the same time, phage AB4P2 exhibited remarkable stability at pH 3-11 and temperatures 30-70 °C. Its dsDNA genome is composed of 45,680 bp with a G + C content of 46.13%. Genomic and phylogenetic analysis situated phage AB4P2 as a new species of Caudoviricetes class. Its fiber protein possesses a pectin lyase-like domain that is linked to depolymerase activity, playing a crucial role in disrupting biofilms. Additionally, it also encodes a lysis cassette comprising endolysin, holin and Rz-like spanin, yet lacks any genes responsible for antibiotic resistance and virulence factors. Phage AB4P2 can completely inhibit A. baumannii growth for 16 h. In the 24-well plate and the polyvinyl chloride (PVC) catheter model experiments, phage AB4P2 achieved a significant biofilm ablation rate and effectively killed the live bacterial cells in the biofilm. CONCLUSIONS Phage AB4P2 had good environmental stability and strong ability to inhibit the growth of A. baumannii and destroy formed biofilms by A. baumannii. It exhibits promising potential for development as an alternative environmental disinfectant against A. baumannii in the hospital. CLINICAL TRIAL NUMBER Not applicable.
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Affiliation(s)
- Jianhui Su
- School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Waihuan West Road 100, Guangzhou City, Guangdong Province, 510006, China
| | - Yujing Tan
- School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Waihuan West Road 100, Guangzhou City, Guangdong Province, 510006, China
| | - Shenshen Liu
- School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Waihuan West Road 100, Guangzhou City, Guangdong Province, 510006, China
| | - Huanhuan Zou
- School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Waihuan West Road 100, Guangzhou City, Guangdong Province, 510006, China
| | - Xiaoyi Huang
- School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Waihuan West Road 100, Guangzhou City, Guangdong Province, 510006, China
| | - Siyi Chen
- School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Waihuan West Road 100, Guangzhou City, Guangdong Province, 510006, China
| | - Hongmei Zhang
- School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Waihuan West Road 100, Guangzhou City, Guangdong Province, 510006, China
| | - Shaoting Li
- School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Waihuan West Road 100, Guangzhou City, Guangdong Province, 510006, China
| | - Haiyan Zeng
- School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Waihuan West Road 100, Guangzhou City, Guangdong Province, 510006, China.
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Lee MM, O'Neil CA, Vogt L, Kwon JH. Environmental hygiene strategies to combat antimicrobial resistance in healthcare settings. ANTIMICROBIAL STEWARDSHIP & HEALTHCARE EPIDEMIOLOGY : ASHE 2025; 5:e71. [PMID: 40109919 PMCID: PMC11920907 DOI: 10.1017/ash.2025.33] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 09/05/2024] [Revised: 01/11/2025] [Accepted: 01/14/2025] [Indexed: 03/22/2025]
Abstract
In this manuscript, we highlight current literature on environmental hygiene techniques to combat reservoirs of antibiotic resistant organisms in the healthcare environment. We discuss several topics for each strategy, including mechanism of action, assessment of effectiveness based on studies, cost, and real-world translatability. The techniques and topics summarized here are not inclusive of all available environmental hygiene techniques but highlight some of the more popular and investigated strategies. We focus on the following: Ultraviolet radiation, hydrogen peroxide vapor, copper-coated surfaces, phages, interventions involving sinks, and educational initiatives.
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Affiliation(s)
- Mary Morgan Lee
- Division of Infectious Diseases, John T. Milliken Department of Internal Medicine, Washington University School of Medicine, St Louis, MO, USA
| | - Caroline A O'Neil
- Division of Infectious Diseases, John T. Milliken Department of Internal Medicine, Washington University School of Medicine, St Louis, MO, USA
| | - Lucy Vogt
- Division of Infectious Diseases, John T. Milliken Department of Internal Medicine, Washington University School of Medicine, St Louis, MO, USA
| | - Jennie H Kwon
- Division of Infectious Diseases, John T. Milliken Department of Internal Medicine, Washington University School of Medicine, St Louis, MO, USA
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Zellner AA, Wirtz DC, Schildberg FA. In Vitro Efficacy of Phage Therapy Against Common Biofilm-forming Pathogens in Orthopedics and Trauma Surgery. ZEITSCHRIFT FUR ORTHOPADIE UND UNFALLCHIRURGIE 2025. [PMID: 39832775 DOI: 10.1055/a-2436-7394] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/22/2025]
Abstract
Formation of biofilms by bacteria is a major challenge in a clinical setting. The importance of these biofilms increases in specialties where foreign bodies and prosthetic material are used. Orthopaedics is such a speciality and phage therapy could offer additional therapeutic options when dealing with biofilm infections.We conducted a systematic literature review using the PubMed database. We searched for phage activity against biofilms of the most common pathogens found in orthopaedics.The results of the systematic review were broken down into different categories and discussed accordingly. We concentrated on the time the biofilms were allowed to mature, and the surface they were grown on. In addition, we checked the efficacy of bacteriophages compared to antibiotics and when applied simultaneously with antibiotics. We also investigated the source of the phages, how they were tested for sensibility against the biofilms, as well the conditions (pH, temperature) under which they remained active and stable.The data suggests that the in vitro efficacy of phages does not change under a wide spectrum of temperature and pH. To further explore the use of bacteriophages in orthopaedics, we need further studies that test biofilms which matured for several weeks on surfaces that are common in arthroplasty and traumatology.
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Affiliation(s)
- Alberto Alfieri Zellner
- Klinik und Poliklinik für Orthopädie und Unfallchirurgie, Universitätsklinikum Bonn, Bonn, Deutschland
| | - Dieter Christian Wirtz
- Klinik und Poliklinik für Orthopädie und Unfallchirurgie, Universitätsklinikum Bonn, Bonn, Deutschland
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Hosseini Hooshiar M, Salari S, Nasiri K, Salim US, Saeed LM, Yasamineh S, Safaralizadeh R. The potential use of bacteriophages as antibacterial agents in dental infection. Virol J 2024; 21:258. [PMID: 39425223 PMCID: PMC11490148 DOI: 10.1186/s12985-024-02510-y] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/13/2024] [Accepted: 09/18/2024] [Indexed: 10/21/2024] Open
Abstract
Dental infections, such as apical Periodontitis, periodontitis, and peri-implantitis (PI), are closely associated with specific bacterial species, including Streptococcus mutans (S. mutans), Porphyromonas gingivalis (P. gingivalis), and Fusobacterium nucleatum (F. nucleatum), among others. Antibiotics are extensively utilized for prophylactic and therapeutic purposes in the treatment of dental infections and other dental-related issues. Unfortunately, the rapid emergence of antimicrobial resistance has accompanied the increased use of antibiotics in recent years. Specific bacterial pathogens have reached a critical stage of antibiotic resistance, characterized by the proliferation of pan-resistant strains and the scarcity of viable therapeutic alternatives. Therapeutic use of particular bacteriophage (phage) particles that target bacterial pathogens is one potential alternative to antibiotics that are now being seriously considered for treating bacterial illnesses. A kind of virus known as a phage is capable of infecting and eliminating bacteria. Because they can't infect cells in plants and animals, phages might be a harmless substitute for antibiotics. To control oral disorders including periodontitis and dental caries, several research have been conducted in this area to study and identify phages from human saliva and dental plaque. The capacity of these agents to disturb biofilms expands their effectiveness against dental plaque biofilms and oral pathogens in cases of periodontitis, PI, and apical periodontitis. This review summarizes the current antibacterial properties of phages used to treat a variety of dental infections, such as periodontitis, peri-implantitis, infected dentin, and apical periodontitis.
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Affiliation(s)
| | - Sara Salari
- Doctor of Dental Surgery, Islamic Azad University of Medical Sciences, Esfahan, Iran
| | - Kamyar Nasiri
- Department of Dentistry, Islamic Azad University of Medical Sciences, Tehran, Iran
| | - Ula Samir Salim
- Department of Dentistry, Al-Noor University College, Nineveh, Iraq
| | - Lamya M Saeed
- Collage of Dentist, National University of Science and Technology, Dhi Qar, 64001, Iraq
| | - Saman Yasamineh
- Young Researchers and Elite Club, Tabriz Branch, Islamic Azad University, Tabriz, Iran.
| | - Reza Safaralizadeh
- Restorative Dentistry Department of Dental Faculty, TABRIZ Medical University, Tabriz, Iran.
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Beck C, Krusche J, Notaro A, Walter A, Kränkel L, Vollert A, Stemmler R, Wittmann J, Schaller M, Slavetinsky C, Mayer C, De Castro C, Peschel A. Wall teichoic acid substitution with glucose governs phage susceptibility of Staphylococcus epidermidis. mBio 2024; 15:e0199023. [PMID: 38470054 PMCID: PMC11005348 DOI: 10.1128/mbio.01990-23] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/26/2023] [Accepted: 02/20/2024] [Indexed: 03/13/2024] Open
Abstract
The species- and clone-specific susceptibility of Staphylococcus cells for bacteriophages is governed by the structures and glycosylation patterns of wall teichoic acid (WTA) glycopolymers. The glycosylation-dependent phage-WTA interactions in the opportunistic pathogen Staphylococcus epidermidis and in other coagulase-negative staphylococci (CoNS) have remained unknown. We report a new S. epidermidis WTA glycosyltransferase TagE whose deletion confers resistance to siphoviruses such as ΦE72 but enables binding of otherwise unbound podoviruses. S. epidermidis glycerolphosphate WTA was found to be modified with glucose in a tagE-dependent manner. TagE is encoded together with the enzymes PgcA and GtaB providing uridine diphosphate-activated glucose. ΦE72 transduced several other CoNS species encoding TagE homologs, suggesting that WTA glycosylation via TagE is a frequent trait among CoNS that permits interspecies horizontal gene transfer. Our study unravels a crucial mechanism of phage-Staphylococcus interaction and horizontal gene transfer, and it will help in the design of anti-staphylococcal phage therapies.IMPORTANCEPhages are highly specific for certain bacterial hosts, and some can transduce DNA even across species boundaries. How phages recognize cognate host cells remains incompletely understood. Phages infecting members of the genus Staphylococcus bind to wall teichoic acid (WTA) glycopolymers with highly variable structures and glycosylation patterns. How WTA is glycosylated in the opportunistic pathogen Staphylococcus epidermidis and in other coagulase-negative staphylococci (CoNS) species has remained unknown. We describe that S. epidermidis glycosylates its WTA backbone with glucose, and we identify a cluster of three genes responsible for glucose activation and transfer to WTA. Their inactivation strongly alters phage susceptibility patterns, yielding resistance to siphoviruses but susceptibility to podoviruses. Many different CoNS species with related glycosylation genes can exchange DNA via siphovirus ΦE72, suggesting that glucose-modified WTA is crucial for interspecies horizontal gene transfer. Our finding will help to develop antibacterial phage therapies and unravel routes of genetic exchange.
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Affiliation(s)
- Christian Beck
- Cluster of Excellence “Controlling Microbes to Fight Infections (CMFI)”, University of Tübingen, Tübingen, Germany
- Interfaculty Institute of Microbiology and Infection Medicine Tübingen, Infection Biology, University of Tübingen, Tübingen, Germany
- German Center for Infection Research (DZIF), Partner Site Tübingen, Tübingen, Germany
| | - Janes Krusche
- Cluster of Excellence “Controlling Microbes to Fight Infections (CMFI)”, University of Tübingen, Tübingen, Germany
- Interfaculty Institute of Microbiology and Infection Medicine Tübingen, Infection Biology, University of Tübingen, Tübingen, Germany
- German Center for Infection Research (DZIF), Partner Site Tübingen, Tübingen, Germany
| | - Anna Notaro
- Department of Agricultural Sciences, University of Naples, Naples, Italy
| | - Axel Walter
- Cluster of Excellence “Controlling Microbes to Fight Infections (CMFI)”, University of Tübingen, Tübingen, Germany
- Interfaculty Institute of Microbiology and Infection Medicine Tübingen, Organismic Interactions/Glycobiology, University of Tübingen, Tübingen, Germany
| | - Lara Kränkel
- Cluster of Excellence “Controlling Microbes to Fight Infections (CMFI)”, University of Tübingen, Tübingen, Germany
- Interfaculty Institute of Microbiology and Infection Medicine Tübingen, Infection Biology, University of Tübingen, Tübingen, Germany
- German Center for Infection Research (DZIF), Partner Site Tübingen, Tübingen, Germany
| | - Anneli Vollert
- Electron-Microscopy, Department of Dermatology, University Hospital Tübingen, Tübingen, Germany
| | - Regine Stemmler
- Cluster of Excellence “Controlling Microbes to Fight Infections (CMFI)”, University of Tübingen, Tübingen, Germany
- Interfaculty Institute of Microbiology and Infection Medicine Tübingen, Infection Biology, University of Tübingen, Tübingen, Germany
- German Center for Infection Research (DZIF), Partner Site Tübingen, Tübingen, Germany
| | - Johannes Wittmann
- Leibniz Institute, DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
| | - Martin Schaller
- Electron-Microscopy, Department of Dermatology, University Hospital Tübingen, Tübingen, Germany
| | - Christoph Slavetinsky
- Cluster of Excellence “Controlling Microbes to Fight Infections (CMFI)”, University of Tübingen, Tübingen, Germany
- Interfaculty Institute of Microbiology and Infection Medicine Tübingen, Infection Biology, University of Tübingen, Tübingen, Germany
- German Center for Infection Research (DZIF), Partner Site Tübingen, Tübingen, Germany
- Pediatric Surgery and Urology, University Children's Hospital Tübingen, University of Tübingen, Tübingen, Germany
| | - Christoph Mayer
- Cluster of Excellence “Controlling Microbes to Fight Infections (CMFI)”, University of Tübingen, Tübingen, Germany
- Interfaculty Institute of Microbiology and Infection Medicine Tübingen, Organismic Interactions/Glycobiology, University of Tübingen, Tübingen, Germany
| | | | - Andreas Peschel
- Cluster of Excellence “Controlling Microbes to Fight Infections (CMFI)”, University of Tübingen, Tübingen, Germany
- Interfaculty Institute of Microbiology and Infection Medicine Tübingen, Infection Biology, University of Tübingen, Tübingen, Germany
- German Center for Infection Research (DZIF), Partner Site Tübingen, Tübingen, Germany
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Novo DC, Edgar KJ. Smart fluorescent polysaccharides: Recent developments and applications. Carbohydr Polym 2024; 324:121471. [PMID: 37985079 PMCID: PMC10661488 DOI: 10.1016/j.carbpol.2023.121471] [Citation(s) in RCA: 5] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/29/2023] [Revised: 09/27/2023] [Accepted: 10/06/2023] [Indexed: 11/22/2023]
Abstract
Polysaccharides are ubiquitous, generally benign in nature, and compatible with many tissues in biomedical situations, making them appealing candidates for new materials such as therapeutic agents and sensors. Fluorescent labeling can create the ability to sensitively monitor distribution and transport of polysaccharide-based materials, which can for example further illuminate drug-delivery mechanisms and therefore improve design of delivery systems. Herein, we review fluorophore selection and ways of appending polysaccharides, utility of the product fluorescent polysaccharides as new smart materials, and their stimulus-responsive nature, with focus on their biomedical applications as environment-sensitive biosensors, imaging, and as molecular rulers. Further, we discuss the advantages and disadvantages of these methods, and future prospects for creation and use of these self-reporting materials.
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Affiliation(s)
- Diana C Novo
- Department of Sustainable Biomaterials, Virginia Tech, Blacksburg, VA 24061, United States; Department of Chemistry, Virginia Tech, Blacksburg, VA 24061, United States
| | - Kevin J Edgar
- Department of Sustainable Biomaterials, Virginia Tech, Blacksburg, VA 24061, United States; Macromolecules Innovation Institute, Virginia Tech, Blacksburg, VA 24061, United States; GlycoMIP, National Science Foundation Materials Innovation Platform, United States.
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7
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Plumet L, Morsli M, Ahmad-Mansour N, Clavijo-Coppens F, Berry L, Sotto A, Lavigne JP, Costechareyre D, Molle V. Isolation and Characterization of New Bacteriophages against Staphylococcal Clinical Isolates from Diabetic Foot Ulcers. Viruses 2023; 15:2287. [PMID: 38140529 PMCID: PMC10747802 DOI: 10.3390/v15122287] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/05/2023] [Revised: 11/09/2023] [Accepted: 11/19/2023] [Indexed: 12/24/2023] Open
Abstract
Staphylococcus sp. is the most common bacterial genus in infections related to diabetic foot ulcers (DFUs). The emergence of multidrug-resistant bacteria places a serious burden on public health systems. Phage therapy is an alternative treatment to antibiotics, overcoming the issue of antibiotic resistance. In this study, six phages (SAVM01 to SAVM06) were isolated from effluents and were used against a panel of staphylococcal clinical samples isolated from DFUs. A genomic analysis revealed that the phages belonged to the Herelleviridae family, with sequences similar to those of the Kayvirus genus. No lysogeny-associated genes, known virulence or drug resistance genes were identified in the phage genomes. The phages displayed a strong lytic and antibiofilm activity against DFU clinical isolates, as well as against opportunistic pathogenic coagulase-negative staphylococci. The results presented here suggest that these phages could be effective biocontrol agents against staphylococcal clinical isolates from DFUs.
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Affiliation(s)
- Lucile Plumet
- VBIC, INSERM U1047, University of Montpellier, 34095 Montpellier, France; (L.P.); (N.A.-M.)
| | - Madjid Morsli
- VBIC, INSERM U1047, Department of Microbiology and Hospital Hygiene, University of Montpellier, CHU Nîmes, 30908 Nîmes, France; (M.M.); (J.-P.L.)
| | - Nour Ahmad-Mansour
- VBIC, INSERM U1047, University of Montpellier, 34095 Montpellier, France; (L.P.); (N.A.-M.)
| | | | - Laurence Berry
- Laboratory of Pathogen and Host Immunity, CNRS UMR5294, University of Montpellier, 34095 Montpellier, France;
| | - Albert Sotto
- VBIC, INSERM U1047, Department of Infectious Diseases, University of de Montpellier, CHU Nîmes, 30908 Nîmes, France;
| | - Jean-Philippe Lavigne
- VBIC, INSERM U1047, Department of Microbiology and Hospital Hygiene, University of Montpellier, CHU Nîmes, 30908 Nîmes, France; (M.M.); (J.-P.L.)
| | | | - Virginie Molle
- VBIC, INSERM U1047, University of Montpellier, 34095 Montpellier, France; (L.P.); (N.A.-M.)
- VBIC, INSERM U1047, Department of Microbiology and Hospital Hygiene, University of Montpellier, CHU Nîmes, 30908 Nîmes, France; (M.M.); (J.-P.L.)
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Fajardo-Lubian A, Venturini C. Use of Bacteriophages to Target Intracellular Pathogens. Clin Infect Dis 2023; 77:S423-S432. [PMID: 37932114 DOI: 10.1093/cid/ciad515] [Citation(s) in RCA: 7] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2023] Open
Abstract
Bacteriophages (phages) have shown great potential as natural antimicrobials against extracellular pathogens (eg, Escherichia coli or Klebsiella pneumoniae), but little is known about how they interact with intracellular targets (eg, Shigella spp., Salmonella spp., Mycobacterium spp.) in the mammalian host. Recent research has demonstrated that phages can enter human cells. However, for the design of successful clinical applications, further investigation is required to define their subcellular behavior and to understand the complex biological processes that underlie the interaction with their bacterial targets. In this review, we summarize the molecular evidence of phage internalization in eucaryotic cells, with specific focus on proof of phage activity against their bacterial targets within the eucaryotic host, and the current proposed strategies to overcome poor penetrance issues that may impact therapeutic use against the most clinically relevant intracellular pathogens.
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Affiliation(s)
- Alicia Fajardo-Lubian
- Faculty of Medicine and Health, Sydney ID Institute, University of Sydney, Sydney, New South Wales, Australia
- Centre for Infectious Diseases and Microbiology, Westmead Institute for Medical Research, Westmead, New South Wales, Australia
| | - Carola Venturini
- Centre for Infectious Diseases and Microbiology, Westmead Institute for Medical Research, Westmead, New South Wales, Australia
- Faculty of Science, Sydney School of Veterinary Science, University of Sydney, Sydney, New South Wales, Australia
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Rajab AAH, Hegazy WAH. What’s old is new again: Insights into diabetic foot microbiome. World J Diabetes 2023; 14:680-704. [PMID: 37383589 PMCID: PMC10294069 DOI: 10.4239/wjd.v14.i6.680] [Citation(s) in RCA: 23] [Impact Index Per Article: 11.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/26/2022] [Revised: 02/20/2023] [Accepted: 04/10/2023] [Indexed: 06/14/2023] Open
Abstract
Diabetes is a chronic disease that is considered one of the most stubborn global health problems that continues to defy the efforts of scientists and physicians. The prevalence of diabetes in the global population continues to grow to alarming levels year after year, causing an increase in the incidence of diabetes complications and health care costs all over the world. One major complication of diabetes is the high susceptibility to infections especially in the lower limbs due to the immunocompromised state of diabetic patients, which is considered a definitive factor in all cases. Diabetic foot infections continue to be one of the most common infections in diabetic patients that are associated with a high risk of serious complications such as bone infection, limb amputations, and life-threatening systemic infections. In this review, we discussed the circumstances associated with the high risk of infection in diabetic patients as well as some of the most commonly isolated pathogens from diabetic foot infections and the related virulence behavior. In addition, we shed light on the different treatment strategies that aim at eradicating the infection.
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Affiliation(s)
- Azza A H Rajab
- Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagzig 44511, Egypt
| | - Wael A H Hegazy
- Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagzig 44511, Egypt
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10
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Štrancar V, Marušić M, Tušar J, Praček N, Kolenc M, Šuster K, Horvat S, Janež N, Peterka M. Isolation and in vitro characterization of novel S. epidermidis phages for therapeutic applications. Front Cell Infect Microbiol 2023; 13:1169135. [PMID: 37293203 PMCID: PMC10244729 DOI: 10.3389/fcimb.2023.1169135] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/18/2023] [Accepted: 05/08/2023] [Indexed: 06/10/2023] Open
Abstract
S. epidermidis is an important opportunistic pathogen causing chronic prosthetic joint infections associated with biofilm growth. Increased tolerance to antibiotic therapy often requires prolonged treatment or revision surgery. Phage therapy is currently used as compassionate use therapy and continues to be evaluated for its viability as adjunctive therapy to antibiotic treatment or as an alternative treatment for infections caused by S. epidermidis to prevent relapses. In the present study, we report the isolation and in vitro characterization of three novel lytic S. epidermidis phages. Their genome content analysis indicated the absence of antibiotic resistance genes and virulence factors. Detailed investigation of the phage preparation indicated the absence of any prophage-related contamination and demonstrated the importance of selecting appropriate hosts for phage development from the outset. The isolated phages infect a high proportion of clinically relevant S. epidermidis strains and several other coagulase-negative species growing both in planktonic culture and as a biofilm. Clinical strains differing in their biofilm phenotype and antibiotic resistance profile were selected to further identify possible mechanisms behind increased tolerance to isolated phages.
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Affiliation(s)
- Vida Štrancar
- Centre of Excellence for Biosensors, Instrumentation and Process Control, Ajdovščina, Slovenia
- Department of Animal Science, Biotechnical Faculty, University of Ljubljana, Domžale, Slovenia
| | - Monika Marušić
- Centre of Excellence for Biosensors, Instrumentation and Process Control, Ajdovščina, Slovenia
| | - Jasmina Tušar
- Centre of Excellence for Biosensors, Instrumentation and Process Control, Ajdovščina, Slovenia
| | - Neža Praček
- Centre of Excellence for Biosensors, Instrumentation and Process Control, Ajdovščina, Slovenia
| | - Marko Kolenc
- Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia
| | - Katja Šuster
- Valdoltra Orthopaedic Hospital, Ankaran, Slovenia
| | - Simon Horvat
- Department of Animal Science, Biotechnical Faculty, University of Ljubljana, Domžale, Slovenia
| | - Nika Janež
- Centre of Excellence for Biosensors, Instrumentation and Process Control, Ajdovščina, Slovenia
| | - Matjaž Peterka
- Centre of Excellence for Biosensors, Instrumentation and Process Control, Ajdovščina, Slovenia
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11
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França A. The Role of Coagulase-Negative Staphylococci Biofilms on Late-Onset Sepsis: Current Challenges and Emerging Diagnostics and Therapies. Antibiotics (Basel) 2023; 12:antibiotics12030554. [PMID: 36978421 PMCID: PMC10044083 DOI: 10.3390/antibiotics12030554] [Citation(s) in RCA: 7] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/31/2022] [Revised: 02/24/2023] [Accepted: 03/06/2023] [Indexed: 03/12/2023] Open
Abstract
Infections are one of the most significant complications of neonates, especially those born preterm, with sepsis as one of the principal causes of mortality. Coagulase-negative staphylococci (CoNS), a group of staphylococcal species that naturally inhabit healthy human skin and mucosa, are the most common cause of late-onset sepsis, especially in preterms. One of the risk factors for the development of CoNS infections is the presence of implanted biomedical devices, which are frequently used for medications and/or nutrient delivery, as they serve as a scaffold for biofilm formation. The major concerns related to CoNS infections have to do with the increasing resistance to multiple antibiotics observed among this bacterial group and biofilm cells’ increased tolerance to antibiotics. As such, the treatment of CoNS biofilm-associated infections with antibiotics is increasingly challenging and considering that antibiotics remain the primary form of treatment, this issue will likely persist in upcoming years. For that reason, the development of innovative and efficient therapeutic measures is of utmost importance. This narrative review assesses the current challenges and emerging diagnostic tools and therapies for the treatment of CoNS biofilm-associated infections, with a special focus on late-onset sepsis.
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Affiliation(s)
- Angela França
- Centre of Biological Engineering, LIBRO—Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal;
- LABBELS—Associate Laboratory in Biotechnology and Bioengineering and Microelectromechanical Systems, Braga and Guimarães, Portugal
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12
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Suh GA, Patel R. Clinical phage microbiology: a narrative summary. Clin Microbiol Infect 2023:S1198-743X(23)00059-9. [PMID: 36805835 DOI: 10.1016/j.cmi.2023.02.006] [Citation(s) in RCA: 6] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/26/2022] [Revised: 02/06/2023] [Accepted: 02/07/2023] [Indexed: 02/17/2023]
Abstract
BACKGROUND Although phage therapy has been in existence for a century, a recent resurgence in interest has occurred because of the continued emergence of antimicrobial resistance and the rising use of indwelling medical devices, resulting in biofilm-associated infections, for which conventional antibiotics are of limited use. Despite this, the clinical successes have been inconsistent because of multiple reasons, including (1) the narrow host range of phages, (2) challenges with concentrating phages at the site of infection, (3) development of resistance of bacteria to phages and (4) immune neutralization. Microbiologic assays have the potential to help guide the course of clinical therapy and improve outcomes. Methods developed decades ago remain the mainstay of phage diagnostics and recently, newer diagnostics are closing the gap needed to further advance clinical phage therapy. OBJECTIVES To review the current state of clinical phage microbiology and identify gaps. SOURCES A PubMed search was performed using the terms "phage microbiology", "phage susceptibility test", "phage host range", "phage biofilm", and "phage therapeutic monitoring". CONTENT Phage susceptibility testing, biofilm assays, phage-antibiotic combination testing, therapeutic drug monitoring, and immune monitoring assays are the current foundation for clinical phage diagnostics. Standardization of these assays and better understanding as to if and how they should be used in terms of clinical management of patients receiving phage therapy is needed. IMPLICATIONS A substantial gap between in vitro studies and in vivo outcomes indicates that further work is needed in phage pharmacokinetics to accurately assay phage particles at the site of infection; recapitulate in vivo biofilm; capture the complex interactions between phages and antibiotics, phages and their target bacteria, among phages in a cocktail, and with the superhost immune system.
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Affiliation(s)
- Gina A Suh
- Division of Public Health, Infectious Diseases, and Occupational Medicine, Department of Medicine, Mayo Clinic, Rochester, MN, USA.
| | - Robin Patel
- Division of Public Health, Infectious Diseases, and Occupational Medicine, Department of Medicine, Mayo Clinic, Rochester, MN, USA; Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA
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13
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Lila ASA, Rajab AAH, Abdallah MH, Rizvi SMD, Moin A, Khafagy ES, Tabrez S, Hegazy WAH. Biofilm Lifestyle in Recurrent Urinary Tract Infections. LIFE (BASEL, SWITZERLAND) 2023; 13:life13010148. [PMID: 36676100 PMCID: PMC9865985 DOI: 10.3390/life13010148] [Citation(s) in RCA: 52] [Impact Index Per Article: 26.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 11/23/2022] [Revised: 12/27/2022] [Accepted: 12/30/2022] [Indexed: 01/06/2023]
Abstract
Urinary tract infections (UTIs) represent one of the most common infections that are frequently encountered in health care facilities. One of the main mechanisms used by bacteria that allows them to survive hostile environments is biofilm formation. Biofilms are closed bacterial communities that offer protection and safe hiding, allowing bacteria to evade host defenses and hide from the reach of antibiotics. Inside biofilm communities, bacteria show an increased rate of horizontal gene transfer and exchange of resistance and virulence genes. Additionally, bacterial communication within the biofilm allows them to orchestrate the expression of virulence genes, which further cements the infestation and increases the invasiveness of the infection. These facts stress the necessity of continuously updating our information and understanding of the etiology, pathogenesis, and eradication methods of this growing public health concern. This review seeks to understand the role of biofilm formation in recurrent urinary tact infections by outlining the mechanisms underlying biofilm formation in different uropathogens, in addition to shedding light on some biofilm eradication strategies.
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Affiliation(s)
- Amr S. Abu Lila
- Department of Pharmaceutics, College of Pharmacy, University of Ha’il, Ha’il 81442, Saudi Arabia
- Molecular Diagnostics and Personalized Therapeutics Unit, University of Ha’il, Ha’il 81442, Saudi Arabia
- Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Zagazig University, Zagazig 44519, Egypt
- Correspondence: (A.S.A.L.); (W.A.H.H.)
| | - Azza A. H. Rajab
- Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagazig 44519, Egypt
| | - Marwa H. Abdallah
- Department of Pharmaceutics, College of Pharmacy, University of Ha’il, Ha’il 81442, Saudi Arabia
- Molecular Diagnostics and Personalized Therapeutics Unit, University of Ha’il, Ha’il 81442, Saudi Arabia
- Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Zagazig University, Zagazig 44519, Egypt
| | - Syed Mohd Danish Rizvi
- Department of Pharmaceutics, College of Pharmacy, University of Ha’il, Ha’il 81442, Saudi Arabia
- Molecular Diagnostics and Personalized Therapeutics Unit, University of Ha’il, Ha’il 81442, Saudi Arabia
| | - Afrasim Moin
- Department of Pharmaceutics, College of Pharmacy, University of Ha’il, Ha’il 81442, Saudi Arabia
- Molecular Diagnostics and Personalized Therapeutics Unit, University of Ha’il, Ha’il 81442, Saudi Arabia
| | - El-Sayed Khafagy
- Department of Pharmaceutics, College of Pharmacy, Prince Sattam bin Abdulaziz University, Al-Kharj 11942, Saudi Arabia
- Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Suez Canal University, Ismailia 41522, Egypt
| | - Shams Tabrez
- King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589, Saudi Arabia
- Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah 21589, Saudi Arabia
| | - Wael A. H. Hegazy
- Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagazig 44519, Egypt
- Pharmacy Program, Department of Pharmaceutical Sciences, Oman College of Health Sciences, Muscat 113, Oman
- Correspondence: (A.S.A.L.); (W.A.H.H.)
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14
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Ramadhan F, Alfiko Y, Purwantomo S, Mubarok AF, Budinarta W, Suwanto A, Budiarti S. A New Approach for Controlling Agrobacterium tumefaciens Post Transformation Using Lytic Bacteriophage. PLANTS (BASEL, SWITZERLAND) 2022; 11:3124. [PMID: 36432853 PMCID: PMC9698577 DOI: 10.3390/plants11223124] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/03/2022] [Revised: 10/26/2022] [Accepted: 11/02/2022] [Indexed: 06/16/2023]
Abstract
Overgrowth of Agrobacterium tumefaciens has frequently been found in Agrobacterium-mediated plant transformation. This overgrowth can reduce transformation efficiency and even lead to explant death. Therefore, this research investigates an alternative way to mitigate or eliminate Agrobacterium after transformation using a bacteriophage. To develop this alternative method, we conducted effectiveness studies of two lytic bacteriophages (ΦK2 and ΦK4) and performed an application test to control Agrobacterium growth after transformation. According to plaque morphological characterization and molecular analysis, the two bacteriophages used in this experiment were distinct. Moreover, some stability physicochemical and growth kinetics, such as adsorption time and susceptibility test, also showed that both bacteriophages differed. On the other hand, the optimum temperature and pH of both phages were the same at 28-30 °C and pH 7. Further investigation showed that both ΦK2 and ΦK4 were able to reduce the overgrowth of A. tumefaciens post transformation. Moreover, applying the cocktail (mixture of ΦK2 and ΦK4) with antibiotic application eradicated A. tumefaciens (0% overgrowth percentage). This result indicates that the application of bacteriophage could be used as an alternative way to eradicate the overgrowth of A. tumefaciens subsequent to transformation.
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Affiliation(s)
- Fiqih Ramadhan
- Graduate School of Biotechnology, IPB University, Bogor 16680, Indonesia
| | - Yuzer Alfiko
- Biotech Laboratory, Wilmar Benih Indonesia, Bekasi 17530, Indonesia
| | - Sigit Purwantomo
- Biotech Laboratory, Wilmar Benih Indonesia, Bekasi 17530, Indonesia
| | | | - Widyah Budinarta
- Biotech Laboratory, Wilmar Benih Indonesia, Bekasi 17530, Indonesia
| | - Antonius Suwanto
- Graduate School of Biotechnology, IPB University, Bogor 16680, Indonesia
- Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Bogor 16680, Indonesia
| | - Sri Budiarti
- Graduate School of Biotechnology, IPB University, Bogor 16680, Indonesia
- Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Bogor 16680, Indonesia
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15
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Antibiofilm activity of a lytic Salmonella phage on different Salmonella enterica serovars isolated from broiler farms. Int Microbiol 2022; 26:205-217. [PMID: 36334144 PMCID: PMC10148789 DOI: 10.1007/s10123-022-00294-1] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/22/2022] [Revised: 10/20/2022] [Accepted: 10/27/2022] [Indexed: 11/06/2022]
Abstract
AbstractBacteriophages have been mainly used in treating infections caused by planktonic bacterial cells in the veterinary sector. However, their applications as antibiofilm agents have received little attention. Accordingly, a previously isolated Salmonella infecting Siphoviridae phage was investigated for host range against 15 Salmonella enterica isolates (S. Cape, S. Gallinarum, 4 S. Enteritidis, 3 S. Montevideo, S. Uno, S. Oritamerin, S. Belgdam, S. Agona, S. Daula, and S. Aba) recovered from the litters of commercial broiler farms. All S. enterica isolates were examined for their biofilm activity using a microtiter plate assay and for adrA, csgD, and gcpA genes using conventional PCR. The phage efficacy against established biofilms produced by the selected seven S. enterica isolates (S. Gallinarum, S. Enteritidis, S. Montevideo, S. Uno, S. Oritamerin, S. Belgdam, and S. Agona) was assessed using microtiter plate assay and reverse transcriptase real-time PCR over different incubation times of 5 and 24 h. All S. enterica isolates were strong biofilm formers. Moreover, the phage effectively reduced the biofilm activity of the established S. enterica biofilms in the microtiter plate assay using the independent sample t-test (P < 0.050). Furthermore, the relative expression levels of csgD, gcpA, and adrA genes in the biofilm cells of S. enterica isolate after phage treatment were significantly up-regulated to variable degrees using the independent sample t-test (P < 0.050). In conclusion, the present study revealed the potential use of Salmonella phage in reducing established biofilms produced by S. enterica serovars isolated from broiler farms.
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16
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Lisac A, Birsa E, Podgornik A. E. coli biofilm formation and its susceptibility towards T4 bacteriophages studied in a continuously operating mixing - tubular bioreactor system. Microb Biotechnol 2022; 15:2450-2463. [PMID: 35638465 PMCID: PMC9437887 DOI: 10.1111/1751-7915.14079] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/02/2022] [Accepted: 05/10/2022] [Indexed: 12/03/2022] Open
Abstract
A system consisting of a connected mixed and tubular bioreactor was designed to study bacterial biofilm formation and the effect of its exposure to bacteriophages under different experimental conditions. The bacterial biofilm inside silicone tubular bioreactor was formed during the continuous pumping of bacterial cells at a constant physiological state for 2 h and subsequent washing with a buffer for 24 h. Monitoring bacterial and bacteriophage concentration along the tubular bioreactor was performed via a piercing method. The presence of biofilm and planktonic cells was demonstrated by combining the piercing method, measurement of planktonic cell concentration at the tubular bioreactor outlet, and optical microscopy. The planktonic cell formation rate was found to be 8.95 × 10-3 h-1 and increased approximately four-fold (4×) after biofilm exposure to an LB medium. Exposure of bacterial biofilm to bacteriophages in the LB medium resulted in a rapid decrease of biofilm and planktonic cell concentration, to below the detection limit within < 2 h. When bacteriophages were supplied in the buffer, only a moderate decrease in the concentration of both bacterial cell types was observed. After biofilm washing with buffer to remove unadsorbed bacteriophages, its exposure to the LB medium (without bacteriophages) resulted in a rapid decrease in bacterial concentration: again below the detection limit in < 2 h.
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Affiliation(s)
- Ana Lisac
- Faculty of Chemistry and Chemical TechnologyUniversity of LjubljanaVečna pot113LjubljanaSlovenia
| | - Elfi Birsa
- Faculty of Chemistry and Chemical TechnologyUniversity of LjubljanaVečna pot113LjubljanaSlovenia
| | - Aleš Podgornik
- Faculty of Chemistry and Chemical TechnologyUniversity of LjubljanaVečna pot113LjubljanaSlovenia
- COBIKMirce 215270AjdovščinaSlovenia
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17
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Šuster K, Cör A. Fast and specific detection of staphylococcal PJI with bacteriophage-based methods within 104 sonicate fluid samples. J Orthop Res 2022; 40:1358-1364. [PMID: 34432330 DOI: 10.1002/jor.25167] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/25/2021] [Revised: 07/19/2021] [Accepted: 07/30/2021] [Indexed: 02/04/2023]
Abstract
The number of prosthetic joint infection (PJI) cases is increasing along with total joint arthroplasties. There is currently no diagnostic test available with 100% sensitivity to identify PJI. The aim of the study was to assess and compare two different bacteriophage K-based methods with standard microbiological culturing methods to detect staphylococci. Samples were retrieved from 104 patients undergoing revision surgery due to suspected PJI. Implants were subjected to sonication and sonicate fluid (SF) was assessed with the methods of qPCR detection of bacteriophage K DNA and adenosine triphosphate (ATP) detection after bacteriophage K lysis. The results were compared with the results of standard microbiological culturing methods. PJI was confirmed in 33 cases according to the PJI definition. Using the methods of ATP and bacteriophage K DNA detection 100% specificity and predictive value were achieved. The sensitivity of qPCR detection was higher (81.25%) than the sensitivity of ATP detection (62.50%) when analyzing SF directly. The sensitivity of the methods significantly improved (to 94.12%) with SF pre-cultivation. Importantly, both methods provided results in 3-4 h when analyzing SF directly, while results from pre-cultivated SF were obtained 19-20 h after sample collection. Our results suggest that bacteriophage-based methods are specific and sensitive and importantly, faster than standard culturing methods. The addition of new bacteriophages to expand the bacterial detection spectrum could lead to the development of a faster, more sensitive, specific, and also economical, and handy method for PJI diagnosis.
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Affiliation(s)
- Katja Šuster
- Department of Research, Valdoltra Orthopaedic Hospital, Ankaran, Slovenia
| | - Andrej Cör
- Department of Research, Valdoltra Orthopaedic Hospital, Ankaran, Slovenia.,Faculty of Education, University of Primorska, Koper, Slovenia
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18
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Visnapuu A, Van der Gucht M, Wagemans J, Lavigne R. Deconstructing the Phage-Bacterial Biofilm Interaction as a Basis to Establish New Antibiofilm Strategies. Viruses 2022; 14:v14051057. [PMID: 35632801 PMCID: PMC9145820 DOI: 10.3390/v14051057] [Citation(s) in RCA: 13] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/01/2021] [Revised: 05/11/2022] [Accepted: 05/11/2022] [Indexed: 12/19/2022] Open
Abstract
The bacterial biofilm constitutes a complex environment that endows the bacterial community within with an ability to cope with biotic and abiotic stresses. Considering the interaction with bacterial viruses, these biofilms contain intrinsic defense mechanisms that protect against phage predation; these mechanisms are driven by physical, structural, and metabolic properties or governed by environment-induced mutations and bacterial diversity. In this regard, horizontal gene transfer can also be a driver of biofilm diversity and some (pro)phages can function as temporary allies in biofilm development. Conversely, as bacterial predators, phages have developed counter mechanisms to overcome the biofilm barrier. We highlight how these natural systems have previously inspired new antibiofilm design strategies, e.g., by utilizing exopolysaccharide degrading enzymes and peptidoglycan hydrolases. Next, we propose new potential approaches including phage-encoded DNases to target extracellular DNA, as well as phage-mediated inhibitors of cellular communication; these examples illustrate the relevance and importance of research aiming to elucidate novel antibiofilm mechanisms contained within the vast set of unknown ORFs from phages.
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19
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Understanding the Mechanisms That Drive Phage Resistance in Staphylococci to Prevent Phage Therapy Failure. Viruses 2022; 14:v14051061. [PMID: 35632803 PMCID: PMC9146914 DOI: 10.3390/v14051061] [Citation(s) in RCA: 18] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/31/2022] [Revised: 05/11/2022] [Accepted: 05/13/2022] [Indexed: 02/07/2023] Open
Abstract
Despite occurring at the microscopic scale, the armed race between phages and their bacterial hosts involves multiple mechanisms, some of which are just starting to be understood. On the one hand, bacteria have evolved strategies that can stop the viral infection at different stages (adsorption, DNA injection and replication, biosynthesis and assembly of the viral progeny and/or release of the newly formed virions); on the other, phages have gradually evolved counterattack strategies that allow them to continue infecting their prey. This co-evolutionary process has played a major role in the development of microbial populations in both natural and man-made environments. Notably, understanding the parameters of this microscopic war will be paramount to fully benefit from the application of phage therapy against dangerous, antibiotic-resistant human pathogens. This review gathers the current knowledge regarding the mechanisms of phage resistance in the Staphylococcus genus, which includes Staphylococcus aureus, one of the most concerning microorganisms in terms of antibiotic resistance acquisition. Some of these strategies involve permanent changes to the bacterial cell via mutations, while others are transient, adaptive changes whose expression depends on certain environmental cues or the growth phase. Finally, we discuss the most plausible strategies to limit the impact of phage resistance on therapy, with a special emphasis on the importance of a rational design of phage cocktails in order to thwart therapeutic failure.
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20
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Abstract
Increasing antimicrobial resistance and medical device-related infections have led to a renewed interest in phage therapy as an alternative or adjunct to conventional antimicrobials. Expanded access and compassionate use cases have risen exponentially but have varied widely in approach, methodology, and clinical situations in which phage therapy might be considered. Large gaps in knowledge contribute to heterogeneity in approach and lack of consensus in many important clinical areas. The Antibacterial Resistance Leadership Group (ARLG) has convened a panel of experts in phage therapy, clinical microbiology, infectious diseases, and pharmacology, who worked with regulatory experts and a funding agency to identify questions based on a clinical framework and divided them into three themes: potential clinical situations in which phage therapy might be considered, laboratory testing, and pharmacokinetic considerations. Suggestions are provided as answers to a series of questions intended to inform clinicians considering experimental phage therapy for patients in their clinical practices.
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21
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Wang L, Pang X, Zhao J, Jin H, Yang X, Fu S, Cheng S, Li H, Miao C, Man C, Jiang Y. Isolation and characteristics of new phage JK004 and application to control Cronobacter sakazakii on material surfaces and powdered infant formula. Lebensm Wiss Technol 2022. [DOI: 10.1016/j.lwt.2021.112571] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
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22
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Mgomi FC, Yuan L, Chen CW, Zhang YS, Yang ZQ. Bacteriophages: A weapon against mixed-species biofilms in the food processing environment. J Appl Microbiol 2021; 133:2107-2121. [PMID: 34932868 DOI: 10.1111/jam.15421] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/04/2021] [Revised: 11/18/2021] [Accepted: 12/17/2021] [Indexed: 11/28/2022]
Abstract
Mixed-species biofilms represent the most frequent actual lifestyles of microorganisms in food processing environments, and they are usually more resistant to control methods than single-species biofilms. The persistence of biofilms formed by foodborne pathogens is believed to cause serious human diseases. These challenges have encouraged researchers to search for novel, natural methods that are more effective towards mixed-species biofilms. Recently, the use of bacteriophages to control mixed-species biofilms have grown significantly in the food industry as an alternative to conventional methods. This review highlights a comprehensive introduction of mixed-species biofilms formed by foodborne pathogens and their enhanced resistance to anti-biofilm removal strategies. Additionally, several methods for controlling mixed-species biofilms briefly focused on applying bacteriophages in the food industry have also been discussed. This article concludes by suggesting that using bacteriophage, combined with other 'green' methods, could effectively control mixed-species biofilms in the food industry.
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Affiliation(s)
- Fedrick C Mgomi
- College of Food Science and Engineering, Yangzhou University, Yangzhou, Jiangsu, 225127, PR China
| | - Lei Yuan
- College of Food Science and Engineering, Yangzhou University, Yangzhou, Jiangsu, 225127, PR China
| | - Cao-Wei Chen
- College of Food Science and Engineering, Yangzhou University, Yangzhou, Jiangsu, 225127, PR China
| | - Yuan-Song Zhang
- College of Food Science and Engineering, Yangzhou University, Yangzhou, Jiangsu, 225127, PR China
| | - Zhen-Quan Yang
- College of Food Science and Engineering, Yangzhou University, Yangzhou, Jiangsu, 225127, PR China
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23
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Biofilm Matrix Formation in Human: Clinical Significance, Diagnostic Techniques, and Therapeutic Drugs. ARCHIVES OF CLINICAL INFECTIOUS DISEASES 2021. [DOI: 10.5812/archcid.107919] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
Abstract
Context: Some recent reports have indicated that almost 80% of clinical infections in humans have biofilm origin and impose additional healthcare costs. This study was an updated review of extracellular polymeric substance matrix (Biofilm) formation in humans and elaborated on its clinical significance, diagnosis, and therapeutic approaches. Evidence Acquisition: This narrative study reviewed the most recent information on the significance of microbial biofilm formation in clinical settings, common biofilm-producing bacterial species, its diagnosis, antibiotic drug resistance, and new approaches to the treatment of infections associated with biofilm formation. Results: Evidence indicated a permanent increase in the frequency of microbial biofilm in the central venous catheter, mechanical heart valve, and urinary catheter, as well as persistent infections. However, antimicrobial resistance induced by biofilms formation and the antimicrobial treatment of biofilms were problematic. Moreover, several assays and lab devices were described to evaluate biofilm formation. Furthermore, new attitudes towards anti-biofilm treatments were introduced in this paper. Conclusions: The number of different mechanisms were in accordance with the recent knowledge on how biofilms play a critical role in the disease pathogenesis. Biofilm strikes the treatment and surveillance of patients bearing infectious diseases under different conditions. The use of new methods in anti-biofilm treatments is effective for the recovery of infected patients.
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24
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Vitale C, Ma TM, Sim J, Altheim C, Martinez-Nieves E, Kadiyala U, Solomon MJ, VanEpps JS. Staphylococcus epidermidis Has Growth Phase Dependent Affinity for Fibrinogen and Resulting Fibrin Clot Elasticity. Front Microbiol 2021; 12:649534. [PMID: 34220741 PMCID: PMC8241941 DOI: 10.3389/fmicb.2021.649534] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/05/2021] [Accepted: 05/21/2021] [Indexed: 11/22/2022] Open
Abstract
Bacterial infection and thrombosis are highly correlated, especially in patients with indwelling medical devices. Coagulase-negative staphylococci, typified by Staphylococcus epidermidis, are a common cause of medical device infections owing to their biofilm forming capacity which provides protection from antibiotics and host immune response. Attention has been drawn to the interaction between S. epidermidis and host proteins, specifically fibrinogen. However, little is known regarding the impact of the transition from planktonic to biofilm forming phenotype on this interaction. Here we investigate the growth phase dependence of bacteria-fibrinogen interaction and the resulting effect on fibrin clot formation, structure, and mechanics. Flow cytometry demonstrated growth phase dependent affinity for fibrinogen. To mimic intravascular device seeding, we quantified the adhesion of S. epidermidis to a fibrinogen coated surface under continuous flow conditions in vitro. The bacterial deposition rate onto fibrinogen was significantly greater for stationary (5,360 ± 1,776 cells/cm2s) versus exponential phase (2,212 ± 264, cells/cm2 s). Furthermore, the expression of sdrG–a cell surface adhesion protein with specificity for fibrinogen–was upregulated ∼twofold in the stationary versus the exponential phase. Rheometry and confocal microscopy demonstrated that stationary phase S. epidermidis slows clot formation and generates a more heterogeneous fibrin network structure with greater elasticity (G′ = 5.7 ± 1.0 Pa) compared to sterile fibrinogen (G′ = l.5 ± 0.2 Pa), while exponential phase cells had little effect. This work contributes to the current understanding of the growth phase dependent regulation of bacterial virulence factors and the correlation between bacterial infection and thrombosis.
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Affiliation(s)
- Carolyn Vitale
- Department of Pediatric Cardiology, University of Michigan, Ann Arbor, MI, United States
| | - Tianhui Maria Ma
- Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, United States
| | - Janice Sim
- Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, United States
| | - Christopher Altheim
- Department of Emergency Medicine, University of Michigan, Ann Arbor, MI, United States
| | - Erika Martinez-Nieves
- Department of Emergency Medicine, University of Michigan, Ann Arbor, MI, United States
| | - Usha Kadiyala
- Department of Emergency Medicine, University of Michigan, Ann Arbor, MI, United States
| | - Michael J Solomon
- Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, United States.,Biointerfaces Institute, University of Michigan, Ann Arbor, MI, United States
| | - J Scott VanEpps
- Department of Emergency Medicine, University of Michigan, Ann Arbor, MI, United States.,Biointerfaces Institute, University of Michigan, Ann Arbor, MI, United States.,Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, United States.,Michigan Center for Integrative Research in Critical Care, University of Michigan, Ann Arbor, MI, United States.,Macromolecular Science and Engineering, University of Michigan, Ann Arbor, MI, United States
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25
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Sartini S, Permana AD, Mitra S, Tareq AM, Salim E, Ahmad I, Harapan H, Emran TB, Nainu F. Current State and Promising Opportunities on Pharmaceutical Approaches in the Treatment of Polymicrobial Diseases. Pathogens 2021; 10:245. [PMID: 33672615 PMCID: PMC7924209 DOI: 10.3390/pathogens10020245] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/10/2021] [Revised: 02/10/2021] [Accepted: 02/18/2021] [Indexed: 12/18/2022] Open
Abstract
In recent years, the emergence of newly identified acute and chronic infectious disorders caused by diverse combinations of pathogens, termed polymicrobial diseases, has had catastrophic consequences for humans. Antimicrobial agents have been clinically proven to be effective in the pharmacological treatment of polymicrobial diseases. Unfortunately, an increasing trend in the emergence of multi-drug-resistant pathogens and limited options for delivery of antimicrobial drugs might seriously impact humans' efforts to combat polymicrobial diseases in the coming decades. New antimicrobial agents with novel mechanism(s) of action and new pharmaceutical formulations or delivery systems to target infected sites are urgently required. In this review, we discuss the prospective use of novel antimicrobial compounds isolated from natural products to treat polymicrobial infections, mainly via mechanisms related to inhibition of biofilm formation. Drug-delivery systems developed to deliver antimicrobial compounds to both intracellular and extracellular pathogens are discussed. We further discuss the effectiveness of several biofilm-targeted delivery strategies to eliminate polymicrobial biofilms. At the end, we review the applications and promising opportunities for various drug-delivery systems, when compared to conventional antimicrobial therapy, as a pharmacological means to treat polymicrobial diseases.
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Affiliation(s)
- Sartini Sartini
- Faculty of Pharmacy, Hasanuddin University, Makassar 90245, Indonesia; (S.S.); (A.D.P.)
| | - Andi Dian Permana
- Faculty of Pharmacy, Hasanuddin University, Makassar 90245, Indonesia; (S.S.); (A.D.P.)
| | - Saikat Mitra
- Department of Pharmacy, Faculty of Pharmacy, University of Dhaka, Dhaka 1000, Bangladesh; or
| | - Abu Montakim Tareq
- Department of Pharmacy, International Islamic University Chittagong, Chittagong 4318, Bangladesh; or
| | - Emil Salim
- Faculty of Pharmacy, Universitas Sumatera Utara, North Sumatera 20155, Indonesia;
| | - Islamudin Ahmad
- Faculty of Pharmacy, Universitas Mulawarman, East Kalimantan 75119, Indonesia;
| | - Harapan Harapan
- Medical Research Unit, School of Medicine, Universitas Syiah Kuala, Banda Aceh 23111, Indonesia;
- Tropical Disease Centre, School of Medicine, Universitas Syiah Kuala, Banda Aceh 23111, Indonesia
- Department of Microbiology, School of Medicine, Universitas Syiah Kuala, Banda Aceh 23111, Indonesia
| | - Talha Bin Emran
- Department of Pharmacy, BGC Trust University Bangladesh, Chittagong 4381, Bangladesh;
| | - Firzan Nainu
- Faculty of Pharmacy, Hasanuddin University, Makassar 90245, Indonesia; (S.S.); (A.D.P.)
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França A, Gaio V, Lopes N, Melo LDR. Virulence Factors in Coagulase-Negative Staphylococci. Pathogens 2021; 10:170. [PMID: 33557202 PMCID: PMC7913919 DOI: 10.3390/pathogens10020170] [Citation(s) in RCA: 79] [Impact Index Per Article: 19.8] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/30/2020] [Revised: 01/29/2021] [Accepted: 01/29/2021] [Indexed: 12/13/2022] Open
Abstract
Coagulase-negative staphylococci (CoNS) have emerged as major pathogens in healthcare-associated facilities, being S. epidermidis, S. haemolyticus and, more recently, S. lugdunensis, the most clinically relevant species. Despite being less virulent than the well-studied pathogen S. aureus, the number of CoNS strains sequenced is constantly increasing and, with that, the number of virulence factors identified in those strains. In this regard, biofilm formation is considered the most important. Besides virulence factors, the presence of several antibiotic-resistance genes identified in CoNS is worrisome and makes treatment very challenging. In this review, we analyzed the different aspects involved in CoNS virulence and their impact on health and food.
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Affiliation(s)
- Angela França
- Laboratory of Research in Biofilms Rosário Oliveira, Centre of Biological Engineering, University of Minho, 4710-057 Braga, Portugal; (V.G.); (N.L.)
| | | | | | - Luís D. R. Melo
- Laboratory of Research in Biofilms Rosário Oliveira, Centre of Biological Engineering, University of Minho, 4710-057 Braga, Portugal; (V.G.); (N.L.)
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Luo Y, Yang Q, Zhang D, Yan W. Mechanisms and Control Strategies of Antibiotic Resistance in Pathological Biofilms. J Microbiol Biotechnol 2021; 31:1-7. [PMID: 33323672 PMCID: PMC9706009 DOI: 10.4014/jmb.2010.10021] [Citation(s) in RCA: 29] [Impact Index Per Article: 7.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/12/2020] [Revised: 11/29/2020] [Accepted: 12/03/2020] [Indexed: 12/15/2022]
Abstract
Bacterial biofilm is a community of bacteria that are embedded and structured in a self-secreted extracellular matrix. An important clinical-related characteristic of bacterial biofilms is that they are much more resistant to antimicrobial agents than the planktonic cells (up to 1,000 times), which is one of the main causes of antibiotic resistance in clinics. Therefore, infections caused by biofilms are notoriously difficult to eradicate, such as lung infection caused by Pseudomonas aeruginosa in cystic fibrosis patients. Understanding the resistance mechanisms of biofilms will provide direct insights into how we overcome such resistance. In this review, we summarize the characteristics of biofilms and chronic infections associated with bacterial biofilms. We examine the current understanding and research progress on the major mechanisms of antibiotic resistance in biofilms, including quorum sensing. We also discuss the potential strategies that may overcome biofilm-related antibiotic resistance, focusing on targeting biofilm EPSs, blocking quorum sensing signaling, and using recombinant phages.
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Affiliation(s)
- Ying Luo
- Department of Pharmacy, Hangzhou Geriatric Hospital, Hangzhou 30022, P.R. China
| | - Qianqian Yang
- Department of Pharmacy, Hangzhou Geriatric Hospital, Hangzhou 30022, P.R. China
| | - Dan Zhang
- Department of Pharmacy, Hangzhou Geriatric Hospital, Hangzhou 30022, P.R. China
| | - Wei Yan
- Department of Pharmacy, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou 310006, P.R. China,Corresponding author Phone/Fax: +86-571-5600-7510 E-mail:
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Pallavi B, Puneeth TG, Shekar M, Girisha SK. Isolation, characterization and genomic analysis of vB-AhyM-AP1, a lytic bacteriophage infecting Aeromonas hydrophila. J Appl Microbiol 2021; 131:695-705. [PMID: 33420733 DOI: 10.1111/jam.14997] [Citation(s) in RCA: 14] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/15/2020] [Revised: 12/28/2020] [Accepted: 01/04/2021] [Indexed: 12/11/2022]
Abstract
AIMS Aeromonas hydrophila is a zoonotic pathogen displaying resistance to multiple antibiotics. Here, we aim to develop a candidate biocontrol agent against A. hydrophila. METHODS AND RESULTS In this study, we isolated and characterized the phage vB-AhyM-AP1 from sewage. It showed lytic activity against A. hydrophila strains. One-step growth curve revealed that the latent period lasted for 40 min. The burst size of one lytic cycle was 1413 PFU per infected cell. Temperature stability studies showed that the phage vB-AhyM-AP1 was active over temperatures ranging from 4 to 45°C for 1 h. pH stability studies indicated that the phage remained active within a pH range of 5-10 after 24 h of incubation. Stability tests in salt solutions showed that the phage was stable at salinities ranging from 0·1 to 2%. The phage also showed stabilities in organic solvents when incubated for 10 min. The Illumina Hiseq sequencing of its genome indicated that the phage vB-AhyM-AP1was a jumbo phage with a genome size of 2, 54 490 bp and GC content of 40·3%. The phylogenetic analysis of the terminase large subunit and major capsid protein indicated that the phage closely clustered with other Tevenvirinae phages. The genome encoded 455 ORFs and 22 tRNAs. The phage resulted in a reduction of 0·8 log units of viable A. hydrophila cells in biofilms grown on PVC coupons maintained in a low nutrient medium for 10 days. CONCLUSIONS The phage showed lytic activity against planktonic and biofilm cells of A. hydrophila. Genome-based prediction showed it to be a strictly lytic phage without any virulence or antibiotic resistance genes indicating safety for environmental and clinical applications. SIGNIFICANCE AND IMPACT OF THE STUDY The multidrug-resistant strains of A. hydrophila pose a significant health risk to both cultured fishes and consumers leaving few options for treatment. Phage vB-AhyM-AP1 may be used as a candidate biocontrol agent against A. hydrophila strains.
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Affiliation(s)
- B Pallavi
- Department of Aquatic Animal Health Management, Karnataka Veterinary, Animal and Fisheries Sciences, University, College of Fisheries, Mangalore, India
| | - T G Puneeth
- Department of Aquatic Animal Health Management, Karnataka Veterinary, Animal and Fisheries Sciences, University, College of Fisheries, Mangalore, India
| | - M Shekar
- Department of Aquatic Animal Health Management, Karnataka Veterinary, Animal and Fisheries Sciences, University, College of Fisheries, Mangalore, India
| | - S K Girisha
- Department of Aquatic Animal Health Management, Karnataka Veterinary, Animal and Fisheries Sciences, University, College of Fisheries, Mangalore, India
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Düzgüneş N, Sessevmez M, Yildirim M. Bacteriophage Therapy of Bacterial Infections: The Rediscovered Frontier. Pharmaceuticals (Basel) 2021; 14:34. [PMID: 33466546 PMCID: PMC7824886 DOI: 10.3390/ph14010034] [Citation(s) in RCA: 24] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/22/2020] [Revised: 12/25/2020] [Accepted: 12/29/2020] [Indexed: 12/18/2022] Open
Abstract
Antibiotic-resistant infections present a serious health concern worldwide. It is estimated that there are 2.8 million antibiotic-resistant infections and 35,000 deaths in the United States every year. Such microorganisms include Acinetobacter, Enterobacterioceae, Pseudomonas, Staphylococcus and Mycobacterium. Alternative treatment methods are, thus, necessary to treat such infections. Bacteriophages are viruses of bacteria. In a lytic infection, the newly formed phage particles lyse the bacterium and continue to infect other bacteria. In the early 20th century, d'Herelle, Bruynoghe and Maisin used bacterium-specific phages to treat bacterial infections. Bacteriophages are being identified, purified and developed as pharmaceutically acceptable macromolecular "drugs," undergoing strict quality control. Phages can be applied topically or delivered by inhalation, orally or parenterally. Some of the major drug-resistant infections that are potential targets of pharmaceutically prepared phages are Pseudomonas aeruginosa, Mycobacterium tuberculosis and Acinetobacter baumannii.
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Affiliation(s)
- Nejat Düzgüneş
- Department of Biomedical Sciences, Arthur A. Dugoni School of Dentistry, University of the Pacific, San Francisco, CA 94103, USA
| | - Melike Sessevmez
- Department of Pharmaceutical Technology, Faculty of Pharmacy, Istanbul University, Istanbul 34116, Turkey;
| | - Metin Yildirim
- Department of Pharmacy Services, Vocational School of Health Services, Tarsus University, Mersin 33400, Turkey;
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Effects of bacteriophage on inhibition and removal of mixed biofilm of enterohemorrhagic Escherichia coli O157:H7 and O91:H-. Lebensm Wiss Technol 2020. [DOI: 10.1016/j.lwt.2020.109945] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/20/2022]
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31
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Rai A, Vittal RV, Mohan Raj JR. Isolation, Characterization, and Comparison of Efficiencies of Bacteriophages to Reduce Planktonic and Biofilm-Associated Staphylococcus aureus. JOURNAL OF HEALTH AND ALLIED SCIENCES NU 2020. [DOI: 10.1055/s-0040-1715773] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/23/2022]
Abstract
Abstract
Introduction In the present era, wherein occurrence of antimicrobial resistance compounded with biofilms in disease conditions has rendered present antibiotic therapy ineffective, the need for alternative strategies to treat bacterial infections has brought bacteriophages to the forefront. The antimicrobial activity of phages is often determined by a viable cell reduction assay which focuses only on planktonic forms. The physiology of an organism in biofilm differs from those that are planktonic; hence, there is a need to evaluate the activity of phages both on planktonic forms, as well as on biofilms, to select candidate therapeutic phages.
Materials and Methods Bacteriophages for Staphylococcus aureus were isolated from environmental samples and characterized based on growth kinetics and DNA fingerprint patterns. Activity of isolated phages on planktonic forms was determined by viable count reduction assay. Phage ability to prevent biofilm formation and ability to disperse formed biofilms were performed in 96-well microtiter plates and biofilm estimated by crystal violet assay.
Results Four bacteriophages designated, that is, P3, PD1, PE1, and PE2, were isolated and characterized. Planktonic cells of S. aureus were found to be sensitive to phages PD1, PE1, and PE2. Phages PD1 and PE2 were efficient in preventing biofilm formation and phages PD1, PE1, and P3 were efficient in dispersing formed biofilms.
Conclusion The ability of some phages to disperse biofilms effectively, while unable to show the same efficiency on planktonic cells, indicates that viable count reduction assay alone may not be a sufficient tool to imply bactericidal activity of bacteriophages, especially while trying to eradicate biofilms.
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Affiliation(s)
- Anoopkrishna Rai
- Division of Infectious Diseases, Nitte University Centre for Science Education and Research, Nitte (Deemed to be University), Deralakatte, Mangaluru, India
| | - Rajeshwari V. Vittal
- Division of Infectious Diseases, Nitte University Centre for Science Education and Research, Nitte (Deemed to be University), Deralakatte, Mangaluru, India
| | - Juliet R. Mohan Raj
- Division of Infectious Diseases, Nitte University Centre for Science Education and Research, Nitte (Deemed to be University), Deralakatte, Mangaluru, India
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Chegini Z, Khoshbayan A, Taati Moghadam M, Farahani I, Jazireian P, Shariati A. Bacteriophage therapy against Pseudomonas aeruginosa biofilms: a review. Ann Clin Microbiol Antimicrob 2020; 19:45. [PMID: 32998720 PMCID: PMC7528332 DOI: 10.1186/s12941-020-00389-5] [Citation(s) in RCA: 149] [Impact Index Per Article: 29.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/07/2020] [Accepted: 09/23/2020] [Indexed: 12/14/2022] Open
Abstract
Multi-Drug Resistant (MDR) Pseudomonas aeruginosa is one of the most important bacterial pathogens that causes infection with a high mortality rate due to resistance to different antibiotics. This bacterium prompts extensive tissue damage with varying factors of virulence, and its biofilm production causes chronic and antibiotic-resistant infections. Therefore, due to the non-applicability of antibiotics for the destruction of P. aeruginosa biofilm, alternative approaches have been considered by researchers, and phage therapy is one of these new therapeutic solutions. Bacteriophages can be used to eradicate P. aeruginosa biofilm by destroying the extracellular matrix, increasing the permeability of antibiotics into the inner layer of biofilm, and inhibiting its formation by stopping the quorum-sensing activity. Furthermore, the combined use of bacteriophages and other compounds with anti-biofilm properties such as nanoparticles, enzymes, and natural products can be of more interest because they invade the biofilm by various mechanisms and can be more effective than the one used alone. On the other hand, the use of bacteriophages for biofilm destruction has some limitations such as limited host range, high-density biofilm, sub-populate phage resistance in biofilm, and inhibition of phage infection via quorum sensing in biofilm. Therefore, in this review, we specifically discuss the use of phage therapy for inhibition of P. aeruginosa biofilm in clinical and in vitro studies to identify different aspects of this treatment for broader use.
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Affiliation(s)
- Zahra Chegini
- Department of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
| | - Amin Khoshbayan
- Student Research Committee, Iran University of Medical Sciences, Tehran, Iran
| | - Majid Taati Moghadam
- Department of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
| | - Iman Farahani
- Molecular and Medicine Research Center, Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran
| | - Parham Jazireian
- Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
| | - Aref Shariati
- Department of Microbiology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
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The Protective Effect of Staphylococcus epidermidis Biofilm Matrix against Phage Predation. Viruses 2020; 12:v12101076. [PMID: 32992766 PMCID: PMC7601396 DOI: 10.3390/v12101076] [Citation(s) in RCA: 21] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/01/2020] [Revised: 09/13/2020] [Accepted: 09/22/2020] [Indexed: 12/12/2022] Open
Abstract
Staphylococcus epidermidis is a major causative agent of nosocomial infections, mainly associated with the use of indwelling devices, on which this bacterium forms structures known as biofilms. Due to biofilms’ high tolerance to antibiotics, virulent bacteriophages were previously tested as novel therapeutic agents. However, several staphylococcal bacteriophages were shown to be inefficient against biofilms. In this study, the previously characterized S. epidermidis-specific Sepunavirus phiIBB-SEP1 (SEP1), which has a broad spectrum and high activity against planktonic cells, was evaluated concerning its efficacy against S. epidermidis biofilms. The in vitro biofilm killing assays demonstrated a reduced activity of the phage. To understand the underlying factors impairing SEP1 inefficacy against biofilms, this phage was tested against distinct planktonic and biofilm-derived bacterial populations. Interestingly, SEP1 was able to lyse planktonic cells in different physiological states, suggesting that the inefficacy for biofilm control resulted from the biofilm 3D structure and the protective effect of the matrix. To assess the impact of the biofilm architecture on phage predation, SEP1 was tested in disrupted biofilms resulting in a 2 orders-of-magnitude reduction in the number of viable cells after 6 h of infection. The interaction between SEP1 and the biofilm matrix was further assessed by the addition of matrix to phage particles. Results showed that the matrix did not inactivate phages nor affected phage adsorption. Moreover, confocal laser scanning microscopy data demonstrated that phage infected cells were less predominant in the biofilm regions where the matrix was more abundant. Our results provide compelling evidence indicating that the biofilm matrix can work as a barrier, allowing the bacteria to be hindered from phage infection.
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Wasfi R, Hamed SM, Amer MA, Fahmy LI. Proteus mirabilis Biofilm: Development and Therapeutic Strategies. Front Cell Infect Microbiol 2020; 10:414. [PMID: 32923408 PMCID: PMC7456845 DOI: 10.3389/fcimb.2020.00414] [Citation(s) in RCA: 80] [Impact Index Per Article: 16.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/28/2020] [Accepted: 07/06/2020] [Indexed: 01/21/2023] Open
Abstract
Proteus mirabilis is a Gram negative bacterium that is a frequent cause of catheter-associated urinary tract infections (CAUTIs). Its ability to cause such infections is mostly related to the formation of biofilms on catheter surfaces. In order to form biofilms, P. mirabilis expresses a number of virulence factors. Such factors may include adhesion proteins, quorum sensing molecules, lipopolysaccharides, efflux pumps, and urease enzyme. A unique feature of P. mirabilis biofilms that build up on catheter surfaces is their crystalline nature owing to their ureolytic biomineralization. This leads to catheter encrustation and blockage and, in most cases, is accompanied by urine retention and ascending UTIs. Bacteria embedded in crystalline biofilms become highly resistant to conventional antimicrobials as well as the immune system. Being refractory to antimicrobial treatment, alternative approaches for eradicating P. mirabilis biofilms have been sought by many studies. The current review focuses on the mechanism by which P. mirabilis biofilms are formed, and a state of the art update on preventing biofilm formation and reduction of mature biofilms. These treatment approaches include natural, and synthetic compounds targeting virulence factors and quorum sensing, beside other strategies that include carrier-mediated diffusion of antimicrobials into biofilm matrix. Bacteriophage therapy has also shown successful results in vitro for combating P. mirabilis biofilms either merely through their lytic effect or by acting as facilitators for antimicrobials diffusion.
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Affiliation(s)
- Reham Wasfi
- Department of Microbiology and Immunology, Faculty of Pharmacy, October University for Modern Sciences and Arts (MSA), Giza, Egypt
| | - Samira M Hamed
- Department of Microbiology and Immunology, Faculty of Pharmacy, October University for Modern Sciences and Arts (MSA), Giza, Egypt
| | - Mai A Amer
- Department of Microbiology and Immunology, Faculty of Pharmacy, October University for Modern Sciences and Arts (MSA), Giza, Egypt
| | - Lamiaa Ismail Fahmy
- Department of Microbiology and Immunology, Faculty of Pharmacy, October University for Modern Sciences and Arts (MSA), Giza, Egypt
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Efficacy of Lytic Phage Cocktails on Staphylococcus aureus and Pseudomonas aeruginosa in Mixed-Species Planktonic Cultures and Biofilms. Viruses 2020; 12:v12050559. [PMID: 32443619 PMCID: PMC7291191 DOI: 10.3390/v12050559] [Citation(s) in RCA: 27] [Impact Index Per Article: 5.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/24/2020] [Revised: 05/11/2020] [Accepted: 05/13/2020] [Indexed: 01/21/2023] Open
Abstract
The efficacy of phages in multispecies infections has been poorly examined. The in vitro lytic efficacies of phage cocktails AB-SA01, AB-PA01, which target Staphylococcus aureus and Pseudomonas aeruginosa, respectively, and their combination against their hosts were evaluated in S. aureus and P. aeruginosa mixed-species planktonic and biofilm cultures. Green fluorescent protein (GFP)-labelled P. aeruginosa PAO1 and mCherry-labelled S. aureus KUB7 laboratory strains and clinical isolates were used as target bacteria. During real-time monitoring using fluorescence spectrophotometry, the density of mCherry S. aureus KUB7 and GFP P. aeruginosa PAO1 significantly decreased when treated by their respective phage cocktail, a mixture of phage cocktails, and gentamicin. The decrease in bacterial density measured by relative fluorescence strongly associated with the decline in bacterial cell counts. This microplate-based mixed-species culture treatment monitoring through spectrophotometry combine reproducibility, rapidity, and ease of management. It is amenable to high-throughput screening for phage cocktail efficacy evaluation. Each phage cocktail, the combination of the two phage cocktails, and tetracycline produced significant biofilm biomass reduction in mixed-species biofilms. This study result shows that these phage cocktails lyse their hosts in the presence of non-susceptible bacteria. These data support the use of phage cocktails therapy in infections with multiple bacterial species.
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36
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Costa SP, Dias NM, Melo LDR, Azeredo J, Santos SB, Carvalho CM. A novel flow cytometry assay based on bacteriophage-derived proteins for Staphylococcus detection in blood. Sci Rep 2020; 10:6260. [PMID: 32277078 PMCID: PMC7148305 DOI: 10.1038/s41598-020-62533-7] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/26/2019] [Accepted: 12/06/2019] [Indexed: 02/08/2023] Open
Abstract
Bloodstream infections (BSIs) are considered a major cause of death worldwide. Staphylococcus spp. are one of the most BSIs prevalent bacteria, classified as high priority due to the increasing multidrug resistant strains. Thus, a fast, specific and sensitive method for detection of these pathogens is of extreme importance. In this study, we have designed a novel assay for detection of Staphylococcus in blood culture samples, which combines the advantages of a phage endolysin cell wall binding domain (CBD) as a specific probe with the accuracy and high-throughput of flow cytometry techniques. In order to select the biorecognition molecule, three different truncations of the C-terminus of Staphylococcus phage endolysin E-LM12, namely the amidase (AMI), SH3 and amidase+SH3 (AMI_SH3) were cloned fused with a green fluorescent protein. From these, a higher binding efficiency to Staphylococcus cells was observed for AMI_SH3, indicating that the amidase domain possibly contributes to a more efficient binding of the SH3 domain. The novel phage endolysin-based flow cytometry assay provided highly reliable and specific detection of 1-5 CFU of Staphylococcus in 10 mL of spiked blood, after 16 hours of enrichment culture. Overall, the method developed herein presents advantages over the standard BSIs diagnostic methods, potentially contributing to an early and effective treatment of BSIs.
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Affiliation(s)
- Susana P Costa
- Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal
- International Iberian Nanotechnology Laboratory (INL), Av. Mestre José Veiga s/n, 4715-330, Braga, Portugal
| | - Nicolina M Dias
- Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal
| | - Luís D R Melo
- Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal
| | - Joana Azeredo
- Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal
| | - Sílvio B Santos
- Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal
| | - Carla M Carvalho
- Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal.
- International Iberian Nanotechnology Laboratory (INL), Av. Mestre José Veiga s/n, 4715-330, Braga, Portugal.
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Walsh DJ, Livinghouse T, Durling GM, Chase-Bayless Y, Arnold AD, Stewart PS. Sulfenate Esters of Simple Phenols Exhibit Enhanced Activity against Biofilms. ACS OMEGA 2020; 5:6010-6020. [PMID: 32226882 PMCID: PMC7098047 DOI: 10.1021/acsomega.9b04392] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 12/20/2019] [Accepted: 02/27/2020] [Indexed: 06/10/2023]
Abstract
The recalcitrance exhibited by microbial biofilms to conventional disinfectants has motivated the development of new chemical strategies to control and eradicate biofilms. The activities of several small phenolic compounds and their trichloromethylsulfenyl ester derivatives were evaluated against planktonic cells and mature biofilms of Staphylococcus epidermidis and Pseudomonas aeruginosa. Some of the phenolic parent compounds are well-studied constituents of plant essential oils, for example, eugenol, menthol, carvacrol, and thymol. The potency of sulfenate ester derivatives was markedly and consistently increased toward both planktonic cells and biofilms. The mean fold difference between the parent and derivative minimum inhibitory concentration against planktonic cells was 44 for S. epidermidis and 16 for P. aeruginosa. The mean fold difference between the parent and derivative biofilm eradication concentration for 22 tested compounds against both S. epidermidis and P. aeruginosa was 3. This work demonstrates the possibilities of a new class of biofilm-targeting disinfectants deploying a sulfenate ester functional group to increase the antimicrobial potency toward microorganisms in biofilms.
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Affiliation(s)
- Danica J Walsh
- Chemistry and Biochemistry, Montana State University, Bozeman, Montana 59717, United States
- Center for Biofilm Engineering, Montana State University, Bozeman, Montana 59717, United States
| | - Tom Livinghouse
- Chemistry and Biochemistry, Montana State University, Bozeman, Montana 59717, United States
| | - Greg M Durling
- Chemistry and Biochemistry, Montana State University, Bozeman, Montana 59717, United States
| | - Yenny Chase-Bayless
- Fish and Wildlife, Montana State University, Bozeman, Montana 59717, United States
| | - Adrienne D Arnold
- Microbiology and Immunology, Montana State University, Bozeman, Montana 59717, United States
| | - Philip S Stewart
- Center for Biofilm Engineering, Montana State University, Bozeman, Montana 59717, United States
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Lewis R, Clooney AG, Stockdale SR, Buttimer C, Draper LA, Ross RP, Hill C. Isolation of a Novel Jumbo Bacteriophage Effective Against Klebsiella aerogenes. Front Med (Lausanne) 2020; 7:67. [PMID: 32185177 PMCID: PMC7058600 DOI: 10.3389/fmed.2020.00067] [Citation(s) in RCA: 17] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/05/2019] [Accepted: 02/13/2020] [Indexed: 12/14/2022] Open
Abstract
Increasing levels of bacterial resistance to many common and last resort antibiotics has increased interest in finding new treatments. The low rate of approval of new antibiotics has led to the search for new and alternative antimicrobial compounds. Bacteriophages (phages) are bacterial viruses found in almost every environment. Phage therapy was historically investigated to control bacterial infections and is still in use in Georgia and as a treatment of last resort. Phage therapy is increasingly recognized as an alternative antimicrobial treatment for antibiotic resistant pathogens. A novel lytic Klebsiella aerogenes phage N1M2 was isolated from maize silage. Klebsiella aerogenes, a member of the ESKAPE bacterial pathogens, is an important target for new antimicrobial therapies. Klebsiella aerogenes can form biofilms on medical devices which aids its environmental persistence and for this reason we tested the effect of phage N1M2 against biofilms. Phage N1M2 successfully removed a pre-formed Klebsiella aerogenes biofilm. Biofilm assays were also carried out with Staphylococcus aureus and Phage K. Phage K successfully removed a preformed Staphylococcus aureus biofilm. Phage N1M2 and Phage K in combination were significantly better at removing a mixed community biofilm of Klebsiella aerogenes and Staphylococcus aureus than either phage alone.
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Affiliation(s)
- Rhea Lewis
- APC Microbiome Ireland, University College Cork, Cork, Ireland.,School of Microbiology, University College Cork, Cork, Ireland
| | - Adam G Clooney
- APC Microbiome Ireland, University College Cork, Cork, Ireland.,School of Microbiology, University College Cork, Cork, Ireland
| | - Stephen R Stockdale
- APC Microbiome Ireland, University College Cork, Cork, Ireland.,School of Microbiology, University College Cork, Cork, Ireland
| | - Colin Buttimer
- APC Microbiome Ireland, University College Cork, Cork, Ireland.,School of Microbiology, University College Cork, Cork, Ireland
| | - Lorraine A Draper
- APC Microbiome Ireland, University College Cork, Cork, Ireland.,School of Microbiology, University College Cork, Cork, Ireland
| | - R Paul Ross
- APC Microbiome Ireland, University College Cork, Cork, Ireland.,School of Microbiology, University College Cork, Cork, Ireland
| | - Colin Hill
- APC Microbiome Ireland, University College Cork, Cork, Ireland.,School of Microbiology, University College Cork, Cork, Ireland
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Adnan M, Ali Shah MR, Jamal M, Jalil F, Andleeb S, Nawaz MA, Pervez S, Hussain T, Shah I, Imran M, Kamil A. Isolation and characterization of bacteriophage to control multidrug-resistant Pseudomonas aeruginosa planktonic cells and biofilm. Biologicals 2019; 63:89-96. [PMID: 31685418 DOI: 10.1016/j.biologicals.2019.10.003] [Citation(s) in RCA: 34] [Impact Index Per Article: 5.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/09/2019] [Revised: 09/05/2019] [Accepted: 10/12/2019] [Indexed: 01/21/2023] Open
Abstract
Pseudomonas aeruginosa is Gram-negative bacterium, one of the leading cause of drug-resistant nosocomial infections in developing countries. This bacterium possesses chromosomally encoded efflux pumps, poor permeability of outer-membrane and high tendency for biofilm formation which are tools to confer resistance. Bacteriophages are regarded as feasible treatment option for control of resistant P. aeruginosa. The aim of the current study was isolate and characterized a bacteriophage against P. aeruginosa with MDR and biofilm ability. A bacteriophage MA-1 with moderate host range was isolated from waste water. The phage was considerable heat and pH stable. Electron microscopy revealed that phage MA-1 belongs to Myoviridae family. Its genome was dsDNA (≈50 kb), coding for eighteen different proteins (ranging from 12 to 250 KDa). P. aeruginosa-2949 log growth phase was significantly reduced by phage MA-1 (2.5 × 103 CFU/ml) as compared to control (without phage). Phage MA-1 also showed significant reductions of 2.0, 2.5 and 3.2 folds in 24, 48, and 74 h old biofilms after 6 h treatment with phage respectively as compared to control. It was concluded from this study that phage MA-1 has capability of killing P. aeruginosa planktonic cells and biofilm, but for complete eradication cocktail will more effective to avoid resistance.
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Affiliation(s)
- Muhammad Adnan
- Department of Biotechnology, Abdul Wali Khan University, Mardan, 23200, Pakistan
| | | | - Muhsin Jamal
- Department of Microbiology, Abdul Wali Khan University, Mardan, 23200, Pakistan.
| | - Fazal Jalil
- Department of Biotechnology, Abdul Wali Khan University, Mardan, 23200, Pakistan
| | - Saadia Andleeb
- Atta-ur-Rahman School of Applied Biosciences, National University of Sciences and Technology, Islamabad, 44000, Pakistan
| | - Muhammad Asif Nawaz
- Department of Biotechnology, Shaheed Benazir Bhutto University, Sheringal, Dir (Upper), Pakistan
| | - Sidra Pervez
- The Karachi Institute of Biotechnology and Genetic Engineering (KIBGE), University of Karachi, Karachi, 75270, Pakistan
| | - Tahir Hussain
- Department of Microbiology, Abdul Wali Khan University, Mardan, 23200, Pakistan
| | - Ismail Shah
- Department of Microbiology, University of Health Sciences, Lahore, Pakistan
| | - Muhammad Imran
- Department of Pharmacy, Abdul Wali Khan University, Mardan, 23200, Pakistan
| | - Atif Kamil
- Department of Biotechnology, Abdul Wali Khan University, Mardan, 23200, Pakistan
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40
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Sharahi JY, Azimi T, Shariati A, Safari H, Tehrani MK, Hashemi A. Advanced strategies for combating bacterial biofilms. J Cell Physiol 2019; 234:14689-14708. [PMID: 30693517 DOI: 10.1002/jcp.28225] [Citation(s) in RCA: 80] [Impact Index Per Article: 13.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/18/2018] [Accepted: 01/16/2019] [Indexed: 01/24/2023]
Abstract
Biofilms are communities of microorganisms that are formed on and attached to living or nonliving surfaces and are surrounded by an extracellular polymeric material. Biofilm formation enjoys several advantages over the pathogens in the colonization process of medical devices and patients' organs. Unlike planktonic cells, biofilms have high intrinsic resistance to antibiotics and sanitizers, and overcoming them is a significant problematic challenge in the medical and food industries. There are no approved treatments to specifically target biofilms. Thus, it is required to study and present innovative and effective methods to combat a bacterial biofilm. In this review, several strategies have been discussed for combating bacterial biofilms to improve healthcare, food safety, and industrial process.
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Affiliation(s)
- Javad Yasbolaghi Sharahi
- Department of Microbiology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Taher Azimi
- Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
| | - Aref Shariati
- Department of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
| | - Hossein Safari
- Health Promotion Research Center, Iran University of Medical Sciences, Tehran, Iran
| | - Melika Khanzadeh Tehrani
- Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
| | - Ali Hashemi
- Department of Microbiology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Abstract
Bacteria in the genus Staphylococcus are important targets for phage therapy due to their prevalence as pathogens and increasing antibiotic resistance. Here we review Staphylococcus outer surface features and specific phage resistance mechanisms that define the host range, the set of strains that an individual phage can potentially infect. Phage infection goes through five distinct phases: attachment, uptake, biosynthesis, assembly, and lysis. Adsorption inhibition, encompassing outer surface teichoic acid receptor alteration, elimination, or occlusion, limits successful phage attachment and entry. Restriction-modification systems (in particular, type I and IV systems), which target phage DNA inside the cell, serve as the major barriers to biosynthesis as well as transduction and horizontal gene transfer between clonal complexes and species. Resistance to late stages of infection occurs through mechanisms such as assembly interference, in which staphylococcal pathogenicity islands siphon away superinfecting phage proteins to package their own DNA. While genes responsible for teichoic acid biosynthesis, capsule, and restriction-modification are found in most Staphylococcus strains, a variety of other host range determinants (e.g., clustered regularly interspaced short palindromic repeats, abortive infection, and superinfection immunity) are sporadic. The fitness costs of phage resistance through teichoic acid structure alteration could make staphylococcal phage therapies promising, but host range prediction is complex because of the large number of genes involved, and the roles of many of these are unknown. In addition, little is known about the genetic determinants that contribute to host range expansion in the phages themselves. Future research must identify host range determinants, characterize resistance development during infection and treatment, and examine population-wide genetic background effects on resistance selection.
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Affiliation(s)
- Abraham G Moller
- Program in Microbiology and Molecular Genetics (MMG), Graduate Division of Biological and Biomedical Sciences (GDBBS), Emory University School of Medicine, Atlanta, Georgia, USA
- Division of Infectious Diseases, Department of Medicine, Emory University School of Medicine, Atlanta, Georgia, USA
| | - Jodi A Lindsay
- Institute of Infection and Immunity, St. George's, University of London, London, United Kingdom
| | - Timothy D Read
- Division of Infectious Diseases, Department of Medicine, Emory University School of Medicine, Atlanta, Georgia, USA
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Dakheel KH, Rahim RA, Neela VK, Al-Obaidi JR, Hun TG, Isa MNM, Yusoff K. Genomic analyses of two novel biofilm-degrading methicillin-resistant Staphylococcus aureus phages. BMC Microbiol 2019; 19:114. [PMID: 31138130 PMCID: PMC6540549 DOI: 10.1186/s12866-019-1484-9] [Citation(s) in RCA: 36] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/02/2018] [Accepted: 05/07/2019] [Indexed: 01/21/2023] Open
Abstract
Background Methicillin-resistant Staphylococcus aureus (MRSA) biofilm producers represent an important etiological agent of many chronic human infections. Antibiotics and host immune responses are largely ineffective against bacteria within biofilms. Alternative actions and novel antimicrobials should be considered. In this context, the use of phages to destroy MRSA biofilms presents an innovative alternative mechanism. Results Twenty-five MRSA biofilm producers were used as substrates to isolate MRSA-specific phages. Despite the difficulties in obtaining an isolate of this phage, two phages (UPMK_1 and UPMK_2) were isolated. Both phages varied in their ability to produce halos around their plaques, host infectivity, one-step growth curves, and electron microscopy features. Furthermore, both phages demonstrated antagonistic infectivity on planktonic cultures. This was validated in an in vitro static biofilm assay (in microtiter-plates), followed by the visualization of the biofilm architecture in situ via confocal laser scanning microscopy before and after phage infection, and further supported by phages genome analysis. The UPMK_1 genome comprised 152,788 bp coding for 155 putative open reading frames (ORFs), and its genome characteristics were between the Myoviridae and Siphoviridae family, though the morphological features confined it more to the Siphoviridae family. The UPMK_2 has 40,955 bp with 62 putative ORFs; morphologically, it presented the features of the Podoviridae though its genome did not show similarity with any of the S. aureus in the Podoviridae family. Both phages possess lytic enzymes that were associated with a high ability to degrade biofilms as shown in the microtiter plate and CLSM analyses. Conclusions The present work addressed the possibility of using phages as potential biocontrol agents for biofilm-producing MRSA. Electronic supplementary material The online version of this article (10.1186/s12866-019-1484-9) contains supplementary material, which is available to authorized users.
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Affiliation(s)
- Khulood Hamid Dakheel
- Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor Darul Ehsan, Malaysia.,Department of Biology, College of Science, Mustansiriyah University, Palestine Street, PO Box 14022, Baghdad, Iraq
| | - Raha Abdul Rahim
- Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor Darul Ehsan, Malaysia.,Institute of Bioscience, Universiti Putra Malaysia, 43400, Serdang, Selangor Darul Ehsan, Malaysia
| | - Vasantha Kumari Neela
- Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor Darul Ehsan, Malaysia
| | - Jameel R Al-Obaidi
- Agro-biotechnology Institute Malaysia (ABI), c/o MARDI Headquarters, 43400, Serdang, Selangor, Malaysia
| | - Tan Geok Hun
- Department of Agriculture Technology, Faculty of Agriculture, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia
| | - Mohd Noor Mat Isa
- Malaysia Genome Institute (MGI), Jalan Bangi, 43000, Kajang, Selangor, Malaysia
| | - Khatijah Yusoff
- Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor Darul Ehsan, Malaysia. .,Institute of Bioscience, Universiti Putra Malaysia, 43400, Serdang, Selangor Darul Ehsan, Malaysia.
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Big Impact of the Tiny: Bacteriophage-Bacteria Interactions in Biofilms. Trends Microbiol 2019; 27:739-752. [PMID: 31128928 DOI: 10.1016/j.tim.2019.04.006] [Citation(s) in RCA: 85] [Impact Index Per Article: 14.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/15/2019] [Revised: 04/10/2019] [Accepted: 04/19/2019] [Indexed: 01/17/2023]
Abstract
Bacteriophages (phages) have been shaping bacterial ecology and evolution for millions of years, for example, by selecting for defence strategies. Evidence supports that bacterial biofilm formation is one such strategy and that biofilm-mediated protection against phage infection depends on maturation and composition of the extracellular matrix. Interestingly, studies have revealed that phages can induce and strengthen biofilms. Here we review interactions between bacteria and phages in biofilms, discuss the underlying mechanisms, the potential of phage therapy for biofilm control, and emphasize the importance of considering biofilms in future phage research. This is especially relevant as biofilms are associated with increased tolerance towards antibiotics and are implicated in the majority of chronic infections.
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Abstract
Staphylococci, with the leading species Staphylococcus aureus and Staphylococcus epidermidis, are the most frequent causes of infections on indwelling medical devices. The biofilm phenotype that those bacteria adopt during device-associated infection facilitates increased resistance to antibiotics and host immune defenses. This review presents and discusses the molecular mechanisms contributing to staphylococcal biofilm development and their in-vivo importance. Furthermore, it summarizes current strategies for the development of therapeutics against staphylococcal biofilm-associated infection.
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Isolation and characterization of a potentially novel Siphoviridae phage (vB_SsapS-104) with lytic activity against Staphylococcus saprophyticus isolated from urinary tract infection. Folia Microbiol (Praha) 2018; 64:283-294. [PMID: 30284669 DOI: 10.1007/s12223-018-0653-9] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/13/2018] [Accepted: 09/25/2018] [Indexed: 02/08/2023]
Abstract
Antibiotic resistance is increasing among Staphylococcus saprophyticus strains isolated from urinary tract infection. This necessitates alternative therapies. For this, a lytic phage (vB_SsapS-104) against S. saprophyticus, which formed round and clear plaques on bacterial culture plates, was isolated from hospital wastewater and characterized. Microscopy analysis showed that it had a small head (about 50 nm), tail (about 80 nm), and a collar (about 22 nm in length and 19 nm in width) indicating to be a phage within Siphoviridae family. Phage vB_SsapS-104 showed a large latency period of about 40 min, rapid adsorption rate that was significantly enhanced by MgCl2 and CaCl2, and high stability to a wide range of temperatures and pH values. Restriction analyses demonstrated that phage consists of a double-stranded DNA with an approximate genome size of 40 Kb. BLAST results did not show high similarity (megablast) with other previously identified phages. But, in Blastn, similarity with Staphylococcus phages was observed. Phage vB_SsapS-104 represented high anti-bacterial activity against S. saprophyticus isolates in vitro as it was able to lyse 8 of the 9 clinical isolates (%88.8) obtained from a hospital in Gorgan, Iran. It was a S. saprophyticus-specific phage because no lytic activity was observed on some other pathogenic bacteria tested. Therefore, phage vB_SsapS-104 can be considered as a specific virulent phage against of S. saprophyitcus isolated from urinary tract infection. This study provided the partial genomic characterization of S. saprophyticus phage and its application against urinary tract infection associated with S. saprophyticus. This phage also can be considered as a good candidate for a therapeutic alternative in the future.
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46
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Jamal M, Andleeb S, Jalil F, Imran M, Nawaz MA, Hussain T, Ali M, Ur Rahman S, Das CR. Isolation, characterization and efficacy of phage MJ2 against biofilm forming multi-drug resistant Enterobacter cloacae. Folia Microbiol (Praha) 2018; 64:101-111. [PMID: 30090964 DOI: 10.1007/s12223-018-0636-x] [Citation(s) in RCA: 31] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/10/2018] [Accepted: 07/31/2018] [Indexed: 11/26/2022]
Abstract
Biofilm is involved in a variety of infections, playing a critical role in the chronicity of infections. Enterobacter cloacae is a biofilm-forming and multi-drug-resistant (MDR) nosocomial pathogen leading to significant morbidity and mortality. This study aimed at isolation of a bacteriophage against MDR clinical strain of E. cloacae and its efficacy against bacterial planktonic cells and biofilm. A bacteriophage MJ2 was successfully isolated from wastewater and was characterized. The phage exhibited a wide range of thermal and pH stability and demonstrated considerable adsorption to host bacteria in the presence of CaCl2 or MgCl2. Transmission electron microscopy (TEM) showed MJ2 head as approximately 62 and 54 nm width and length, respectively. It had a short non-contractile tail and was characterized as a member of the family Podoviridae [order Caudovirales]. The phage MJ2 was found to possess 11 structural proteins (12-150 kDa) and a double-stranded DNA genome with an approximate size of 40 kb. The log-phase growth of E. cloacae both in biofilm and suspension was significantly reduced by the phage. The E. cloacae biofilm was formed under different conditions to evaluate the efficacy of MJ2 phage. Variable reduction pattern of E. cloacae biofilm was observed while treating it for 4 h with MJ2, i.e., biofilm under static conditions. The renewed media with intervals of 24, 72, and 120 h showed biomass decline of 2.8-, 3-, and 3.5-log, respectively. Whereas, the bacterial biofilm formed with dynamic conditions with refreshing media after 24, 72, and 120 h demonstrated decline in growth at 2.5-, 2.6-, and 3.3-log, respectively. It was, therefore, concluded that phage MJ2 possessed considerable inhibitory effects on MDR E. cloacae both in planktonic and biofilm forms.
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Affiliation(s)
- Muhsin Jamal
- Department of Microbiology, Abdul Wali Khan University, Garden Campus, Mardan, 23200, Pakistan.
- Emerging Pathogens Institute (EPI), University of Florida (UF), Gainesville, FL, USA.
- College of Veterinary Sciences & Animal Husbandry, Abdul Wali Khan University, Mardan, Pakistan.
| | - Saadia Andleeb
- Emerging Pathogens Institute (EPI), University of Florida (UF), Gainesville, FL, USA
- Atta-ur-Rahman School of Applied Biosciences, National University of Sciences and Technology, Islamabad, Pakistan
| | - Fazal Jalil
- Atta-ur-Rahman School of Applied Biosciences, National University of Sciences and Technology, Islamabad, Pakistan
- Department of Biotechnology, Abdul Wali Khan University, Mardan, 23200, Pakistan
| | - Muhammad Imran
- Department of Biotechnology, Abdul Wali Khan University, Mardan, 23200, Pakistan
- Department of Microbiology, University of Health Sciences, Lahore, Pakistan
| | - Muhammad Asif Nawaz
- Department of Microbiology, University of Health Sciences, Lahore, Pakistan
- Department of Biotechnology, Shaheed Benazir Bhutto University, Sheringal, Dir (Upper), Pakistan
| | - Tahir Hussain
- Department of Microbiology, Abdul Wali Khan University, Garden Campus, Mardan, 23200, Pakistan
| | - Muhammad Ali
- Department of Biotechnology, Shaheed Benazir Bhutto University, Sheringal, Dir (Upper), Pakistan
- Department of Life Sciences, School of Science, University of Management and Technology, C-II Johar Town, Lahore, 54770, Pakistan
| | - Sadeeq Ur Rahman
- College of Veterinary Sciences & Animal Husbandry, Abdul Wali Khan University, Mardan, Pakistan
- Department of Life Sciences, School of Science, University of Management and Technology, C-II Johar Town, Lahore, 54770, Pakistan
| | - Chythanya Rajanna Das
- Emerging Pathogens Institute (EPI), University of Florida (UF), Gainesville, FL, USA
- College of Veterinary Sciences & Animal Husbandry, Abdul Wali Khan University, Mardan, Pakistan
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Melo LDR, França A, Brandão A, Sillankorva S, Cerca N, Azeredo J. Assessment of Sep1virus interaction with stationary cultures by transcriptional and flow cytometry studies. FEMS Microbiol Ecol 2018; 94:5061119. [DOI: 10.1093/femsec/fiy143] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/28/2018] [Accepted: 07/26/2018] [Indexed: 12/24/2022] Open
Affiliation(s)
- Luís D R Melo
- CEB - Centre of Biological Engineering, LIBRO - Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, 4710-057 Braga, Portugal
| | - Angela França
- CEB - Centre of Biological Engineering, LIBRO - Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, 4710-057 Braga, Portugal
| | - Ana Brandão
- CEB - Centre of Biological Engineering, LIBRO - Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, 4710-057 Braga, Portugal
| | - Sanna Sillankorva
- CEB - Centre of Biological Engineering, LIBRO - Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, 4710-057 Braga, Portugal
| | - Nuno Cerca
- CEB - Centre of Biological Engineering, LIBRO - Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, 4710-057 Braga, Portugal
| | - Joana Azeredo
- CEB - Centre of Biological Engineering, LIBRO - Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, 4710-057 Braga, Portugal
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Yazdi M, Bouzari M, Ghaemi E. Isolation and Characterization of a Lytic Bacteriophage (vB_PmiS-TH) and Its Application in Combination with Ampicillin against Planktonic and Biofilm Forms of Proteus mirabilis Isolated from Urinary Tract Infection. J Mol Microbiol Biotechnol 2018; 28:37-46. [DOI: 10.1159/000487137] [Citation(s) in RCA: 26] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/09/2017] [Accepted: 01/22/2018] [Indexed: 11/19/2022] Open
Abstract
<i>Proteus mirabilis</i> is one of the most common causes of urinary tract infection (UTI), particularly in patients undergoing long-term catheterization. Phage vB_PmiS-TH was isolated from wastewater with high lytic activity against <i>P. mirabilis</i> (TH) isolated from UTI. The phage had rapid adsorption, a large burst size (∼260 PFU per infected cell), and high stability at a wide range of temperatures and pH values. As analyzed by transmission electron microscopy, phage vB_PmiS-TH had an icosahedral head of ∼87 × 62 nm with a noncontractile tail about 137 nm in length and 11 nm in width. It belongs to the family <i>Siphoviridae</i>. Combination of the phage vB_PmiS-TH with ampicillin had a higher removal activity against planktonic cells of <i>P. mirabilis</i> (TH) than the phage or the antibiotic alone. Combination of the phage at a multiplicity of infection of 100 with a high dose of ampicillin (246 µg/mL) showed the highest biofilm removal activity after 24 h. This study demonstrates that using a combination of phage and antibiotic could be significantly more effective against planktonic and biofilm forms of <i>P. mirabilis</i> (TH).
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49
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Anjum S, Singh S, Benedicte L, Roger P, Panigrahi M, Gupta B. Biomodification Strategies for the Development of Antimicrobial Urinary Catheters: Overview and Advances. GLOBAL CHALLENGES (HOBOKEN, NJ) 2018; 2:1700068. [PMID: 31565299 PMCID: PMC6607219 DOI: 10.1002/gch2.201700068] [Citation(s) in RCA: 29] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 08/14/2017] [Revised: 10/05/2017] [Indexed: 05/27/2023]
Abstract
Microbial burden associated with medical devices poses serious health challenges and is accountable for an increased number of deaths leading to enormous medical costs. Catheter-associated urinary tract infections are the most common hospital-acquired infections with enhanced patient morbidity. Quite often, catheter-associated bacteriuria produces apparent adverse outcomes such as urosepsis and even death. Taking this into account, the methods to modify urinary catheters to control microbial infections with relevance to clinical drug resistance are systematically evaluated in this review. Technologies to restrict biofilm formation at initial stages by using functional nanomaterials are elucidated. The conventional methodology of using single therapeutic intervention for developing an antimicrobial catheter lacks clinically meaningful benefit. Therefore, catheter modification using naturally derived antimicrobials such as essential oils, curcumin, enzymes, and antimicrobial peptides in combination with synthetic antibiotics/nanoantibiotics is likely to exert sufficient inhibitory effect on uropathogens and is extensively discussed. Futuristic efforts in this area are projected here that demand clinical studies to address areas of uncertainty to avoid development of bacterial resistance to the new generation therapy with minimum discomfort to the patients.
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Affiliation(s)
- Sadiya Anjum
- Bioengineering LaboratoryDepartment of Textile TechnologyIndian Institute of TechnologyNew Delhi110016India
| | - Surabhi Singh
- Bioengineering LaboratoryDepartment of Textile TechnologyIndian Institute of TechnologyNew Delhi110016India
| | - Lepoittevin Benedicte
- ICMMO ‐ LG2M ‐ Bât 420Université Paris‐Sud XI, 15rue Georges Clémenceau91405Orsay CedexFrance
| | - Philippe Roger
- ICMMO ‐ LG2M ‐ Bât 420Université Paris‐Sud XI, 15rue Georges Clémenceau91405Orsay CedexFrance
| | - Manoj Panigrahi
- Department of Urology and PathologySikkim Manipal Institute of Medical SciencesGangtokSikkim737101India
| | - Bhuvanesh Gupta
- Bioengineering LaboratoryDepartment of Textile TechnologyIndian Institute of TechnologyNew Delhi110016India
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50
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Abedon ST. Bacteriophage-Mediated Biocontrol of Wound Infections, and Ecological Exploitation of Biofilms by Phages. BIOFILM, PILONIDAL CYSTS AND SINUSES 2018. [DOI: 10.1007/15695_2018_110] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
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