bims-fagtap Biomed News
on Phage therapies and applications
Issue of 2025–05–11
fifteen papers selected by
Luca Bolliger, lxBio
  1. [Phage therapy in Germany-Update 2025].
  2. Pharmacokinetics and pharmacodynamics studies of phage therapy.
  3. [Medical ethical challenges of phage therapy-informed consent, study design and therapeutic trial].
  4. Infected burn wound healing using Hydroxy-propyl-methyl cellulose gel containing bacteriophages against Pseudomonas aeruginosa and Klebsiella pneumoniae.
  5. [Phage therapeutics and pharmaceutical legislation: classification, manufacture, placing on the market, and application].
  6. [Phage endolysins-a novel class of antibacterial agents with a wide range of applications].
  7. Bacteriophages Improve the Effectiveness of Rhamnolipids in Combating the Biofilm of Candida albicans.
  8. Engineering artificial microbial consortia for personalized gut microbiome modulation and disease treatment.
  9. Bacteriophage-based gene delivery: a novel approach for targeted breast cancer therapy.
  10. From resistance to treatment: the ongoing struggle with Acinetobacter baumannii.
  11. Evaluating the Antibiofilm Effects of Antibiotics on Staphylococcus Species from Pediatric Hematology-Oncology Patients.
  12. A review on smart dressings with advanced features.
  13. Development of a Korean Nutrition Model for In Silico Gut Microbiome Analyses Integrated With Nutrigenomics.
  14. Resistance profiles of Staphylococcus aureus isolates against frequently used antibiotics at private sector laboratories in Jordan.
  15. Cystic Fibrosis Aggregate Biofilm Model to Study Infection-relevant Gene Expression.
  1. Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz. 2025 May 08.
    [Phage therapy in Germany-Update 2025].
    Christian Willy, Felix Bröcker.
      Phage therapy is a promising approach to combat the antibiotic resistance crisis worldwide and in Germany. However, after widespread use in Germany from the 1930s to the 1960s, the number of patients treated with phages each year dropped to under a hundred in the following decades. An overview of the current situation of phage therapy in Germany and the translational research supporting it is provided. Aspects taken into account here are phage production, phage banks, clinical phage application, and ongoing translational research projects in Germany.There is a lack of phages for clinical use. Phage therapy is only used in a limited number of patients in a few clinics as an individual treatment attempt. There are several phage banks that, due to different institutional backgrounds and guidelines, do not work in a comparable way and only exchange phage stocks in the context of scientific projects. Projects in basic and translational phage research have increased considerably in Germany. The most urgent change seems to be the expansion of phage production capacities.There is still no high-quality evidence on the clinical success of phage therapy in the medical field by means of randomized controlled clinical trials. In close consultation with the regulatory authorities, it seems sensible to urgently enable a few centers to treat patients according to the Belgian model.
    Keywords:  Germany; Phage bank; Phage production; Phage therapy; Research projects
    DOI:  https://doi.org/10.1007/s00103-025-04063-z
  2. Farm Hosp. 2025 May 08. pii: S1130-6343(25)00052-2. [Epub ahead of print]
    Pharmacokinetics and pharmacodynamics studies of phage therapy.
    Lucía Blasco, Inés Bleriot, Patricia Fernández-Grela, José Ramón Paño-Pardo, Jesús Oteo-Iglesias, María Tomás.
      The need for new antimicrobial treatments that work alternatively or synergistically with antibiotics to address the problem of the emergence and transmission of antimicrobial resistance has increased interest in the use of minority therapies such as phage therapy. For safe and widespread application of this therapy, it is necessary to establish the pharmacokinetic and pharmacodynamic parameters for its use in humans. This systematic review analyzes the criteria necessary to establish the PK/PD of this therapy, as well as its current application, based on a review of 66 clinical cases that catch diverse infections and phage administration routes.
    Keywords:  Efficacy; Eficacia; Fagos; Fagoterapia; Farmacocinéticos y farmacodinámicos; Phage therapy; Phages; Pharmacokinetics and pharmacodynamics; Sinergias; Synergies
    DOI:  https://doi.org/10.1016/j.farma.2025.04.003
  3. Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz. 2025 May 06.
    [Medical ethical challenges of phage therapy-informed consent, study design and therapeutic trial].
    Hans Zillmann.
      The antibiotics crisis is increasing interest in bacteriophages and related therapeutic approaches. The translation of phage therapy into clinical application faces various ethical challenges. These include the patient's informed consent based on patient information as well as open questions regarding an ethically sound study design. In addition, further ethical problems arise from the current situation of phage therapy in Germany with regard to equity of access. This article reflects on the three aspects of phage therapy mentioned above. The results show that phage therapy does not pose any requirements for patient information that could not be solved by adapting the information provided. Questions regarding an adapted study design arise against the background of the placebo dilemma, possible resistance to antibiotics and the expected small number of study participants. An attempt can be made to respond to these challenges by adapting the study design. As a result, the studies deviate from the gold standard, which is randomized, controlled and double-blind studies. A balance must be struck here between evidence requirements and the protection of study participants. From an ethical point of view, the protection of participants must be given greater weight. The current situation of phage therapy in Germany poses an ethical problem, as both the individualised therapeutic trial and the resulting medical tourism that can be observed stand in the way of the demand for fair access and distribution. In order to overcome this status, the magistral prescription represents a possible interim step until clarity has been gained about an appropriate study design.
    Keywords:  Autonomy; Bacteriophages; Justice; Medical ethics; Translation
    DOI:  https://doi.org/10.1007/s00103-025-04056-y
  4. Iran J Microbiol. 2025 Feb;17(1): 69-79
    Infected burn wound healing using Hydroxy-propyl-methyl cellulose gel containing bacteriophages against Pseudomonas aeruginosa and Klebsiella pneumoniae.
    Meysam Adibi, Kazem Javanmardi, Noor Al-Huda Ali A H Saeed, Mohammad Ali Mobasher, Javad Jokar, Abdolmajid Ghasemian, Niloofar Rahimian, Ava Soltani Hekmat.
       Background and Objectives: Pseudomonas aeruginosa (P. aeruginosa) and Klebsiella pneumoniae (K. pneumoniae) are the two leading bacterial strains involved in wound infections. These bacteria have developed broad resistance to antibiotics, which has complicated their eradication. Additionally, the formation of a polymicrobial infection poses additional problems. Among alternative or complementary options, bacteriophages, viruses that parasitize bacterial hosts, have been promising.
    Materials and Methods: In this research work, bacteriophages' therapeutic effects against P. aeruginosa- and K. pneumoniae-infected burn wounds were studied. The infectious burn wound model was performed on Balb/C male mice, aged six weeks and weighing 25 ± 5 gr. The effects of the Hydroxy-propyl-methyl cellulose (HPMC) gel containing phage were investigated compared to gentamicin. All of these actions were performed in separate groups for each bacteria and mixed group of bacteria (to test multi-bacterial infections treating) and the result were compared.
    Results: Phages appear to be effective in gel forms. Pathologic samples of different groups confirmed therapeutic results of phages. These results at the microscopic level indicated the recovery of the tissue and the removal of the infection.
    Conclusion: The results of this study indicate that lytic phages are powerful biological tools for the treatment of bacterial infections in burn wounds, which can be considered as one of the alternatives for drug-resistant bacterial species and the high costs of antibiotics; though further animal and trial studies are needed. Meanwhile, the complications due to their widespread use in humans should be investigated in more details.
    Keywords:  Antibiotic resistance; Bacteriophages; Hydroxy-propyl-methyl cellulose
    DOI:  https://doi.org/10.18502/ijm.v17i1.17803
  5. Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz. 2025 May 09.
    [Phage therapeutics and pharmaceutical legislation: classification, manufacture, placing on the market, and application].
    Timo Faltus.
      Currently, most phage therapeutics are individualized, that is, prepared specifically for a single patient. In this case, the preparation is usually decentralized and takes place in the respective hospital pharmacies as extemporaneous preparations. The technical supply of phage medicinal products thus differs from the guiding principle of pharmaceutical law. The assurance of quality, efficacy, and safety of medicinal products is typically adapted to the supply of medicinal products that are centrally manufactured independently of the patient in advance and administered decentrally. Only in legally defined exceptional cases may medicinal products be prepared decentrally by pharmacies or physicians without the need for a manufacturing authorization or a marketing authorization prior to application.The article shows why, from a legal perspective, phage medicinal products based on wild-type phages are simple, non-specifically classified medicinal product by virtue of function. Based on this, the article describes the legal requirements under which individualized and non-individualized phage medicinal products can be manufactured, placed on the market and administered. The article also addresses legal issues relating to phage lysins. Furthermore, the article shows the impact of the legal handling of phage pharmaceuticals on the requirements for the reimbursement of phage pharmaceuticals by health insurance providers. In view of the technical requirements of phage therapy, the article discusses the extent to which the current legal framework enables the translation of phage medicinal products into standard medical care. Finally, the article provides an overview of currently planned legislative changes and their likely impact on the legal handling of phage medicinal products.
    Keywords:  Extemporaneous preparation; Hospital pharmacy; Personalized medicine; Phage lysins; Phage therapy medicinal products
    DOI:  https://doi.org/10.1007/s00103-025-04054-0
  6. Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz. 2025 May 06.
    [Phage endolysins-a novel class of antibacterial agents with a wide range of applications].
    Evgeny A Idelevich, Karsten Becker.
      As "enzybiotics," endolysins represent a new class of antibacterial agents that are naturally produced at the end of the lytic cycle in bacteriophage-infected bacterial cells to enable the phage particles formed to be released from the inside of the host cell. Their enzymatic effect on the cell wall peptidoglycan, which leads to lysis of the infected bacteria, can also be exerted externally as an applied agent. While the endolysin activity can be directly effective in Gram-positive bacteria, the endolysin must be modified for activity against Gram-negative bacteria so that it can overcome the outer cell membrane. For this reason, and to optimize endolysin specificity and stability, endolysins are increasingly being genetically modified and produced recombinantly, which is relatively easy to achieve due to their modular structure consisting of lytic and binding domains. Endolysins have already found increasing actual or extensively postulated use for preventive, therapeutic, and diagnostic purposes in human and veterinary medicine as well as in food safety, biotechnology, and the One Health sector; however, this still needs to be better substantiated by valid studies. Although, in contrast to phage therapy, the regulatory aspects can follow the approval procedures also required for other pharmaceuticals, only less than a dozen randomized controlled studies of phases 1 to 3 have been initiated or completed in the field of human medicine. Only very few endolysin formulations approved as medical products are currently available on the market and approval as medicinal drugs is being sought for some endolysins.
    Keywords:  Bacteriophage; Endolysin; Infection; Prevention; Therapy
    DOI:  https://doi.org/10.1007/s00103-025-04059-9
  7. Molecules. 2025 Apr 15. pii: 1772. [Epub ahead of print]30(8):
    Bacteriophages Improve the Effectiveness of Rhamnolipids in Combating the Biofilm of Candida albicans.
    Izabela Dusza, Dominika Jama, Grzegorz Skaradziński, Paulina Śliwka, Tomasz Janek, Aneta Skaradzińska.
      Biofilms formed by Candida albicans pose therapeutic challenges due to their resistance to conventional antimicrobials, highlighting the need for more effective treatments. Rhamnolipids (RLs) are biosurfactants with diverse antimicrobial properties. Bacteriophages are viruses that target specific bacterial strains. Recent studies have shown that they may affect biofilm formation by fungi and yeasts. This study investigated the combined antimicrobial effects of RLs and bacteriophages against C. albicans biofilms, focusing on their anti-adhesive and inhibitory effects on biofilm development. RT-PCR assays were used to analyze gene modulation in C. albicans biofilm formation in response to RLs and bacteriophage treatments, while hyphae formation was examined using microscopy. The results showed that RLs-bacteriophage combinations significantly reduced biofilm formation compared to individual treatments. A combination of 200 mg/L RLs with bacteriophage BF9 led to a 94.8% reduction in biofilm formation. In a subsequent model, the same RL concentration with bacteriophage LO5/1f nearly eliminated biofilm formation (~96%). Gene expression analysis revealed downregulation of key biofilm-associated genes when Candida cells were treated with 200 mg/L RLs and four bacteriophages (BF17, LO5/1f, JG004, FD). These results show the potential of RL and bacteriophage combinations in combating C. albicans biofilms, presenting a promising therapeutic approach against resilient infections.
    Keywords:  Candida albicans; antimicrobial agents; bacteriophages; biofilm assay; combined therapy; rhamnolipids
    DOI:  https://doi.org/10.3390/molecules30081772
  8. Ann N Y Acad Sci. 2025 May 05.
    Engineering artificial microbial consortia for personalized gut microbiome modulation and disease treatment.
    Timoth Mkilima.
      The human gut microbiome is a complex ecosystem that plays a vital role in maintaining health and contributing to the pathogenesis of various diseases. This review proposes a transformative approach that involves engineering artificial microbial consortia-precisely designed communities of microorganisms-for personalized modulation of the gut microbiome and targeted therapeutic interventions. By integrating synthetic biology, systems biology, and advanced culturing techniques, tailored microbial consortia can be developed to perform specific functions within the gut, including the production of therapeutic molecules, modulation of immune responses, and competition against pathogenic bacteria. In vitro and in vivo studies indicate that these engineered consortia can effectively restore microbial balance and enhance host resilience. This personalized approach holds immense potential to revolutionize healthcare by addressing the root causes of diseases such as metabolic disorders, inflammatory conditions, and gastrointestinal infections through precise manipulation of the gut microbiome. Future research should focus on rigorous clinical trials to evaluate the safety, efficacy, and long-term impacts of these engineered consortia in diverse human populations, paving the way for innovative microbial therapies that promote overall health and well-being.
    Keywords:  gut microbiome engineering; microbial consortia; personalized medicine; synthetic biology; therapeutic microbiology
    DOI:  https://doi.org/10.1111/nyas.15352
  9. Nucleosides Nucleotides Nucleic Acids. 2025 May 05. 1-19
    Bacteriophage-based gene delivery: a novel approach for targeted breast cancer therapy.
    Dilpreet Singh.
      Bacteriophage-based gene delivery systems are emerging as a promising alternative to traditional viral and non-viral vectors for targeted gene therapy in breast cancer. Their unique structural adaptability, low immunogenicity, and cost-effective production make them ideal candidates for precision medicine applications. Unlike conventional gene delivery platforms, bioengineered bacteriophages can be functionalized with tumor-specific ligands, modified for PEGylation to enhance circulation stability, and integrated with CRISPR/Cas9 gene-editing systems for precise genomic modifications. Additionally, bacteriophage vectors can be utilized in combination therapy, amplifying the effectiveness of chemotherapy and immunotherapy in breast cancer treatment. This mini-review discusses the bioengineering strategies used to enhance bacteriophage-based gene delivery, including surface modifications for tumor targeting, ligand-receptor binding for cellular uptake, and controlled genetic cargo release. We further examine in vitro and in vivo studies that demonstrate the potential of bacteriophage vectors in tumor suppression, gene expression efficiency, and immunomodulation. Furthermore, we explore the challenges and future directions of integrating bacteriophage-mediated gene therapy into clinical applications, addressing key issues such as systemic circulation half-life, off-target effects, and immune system interactions.
    Keywords:  Bacteriophage-based gene delivery; CRISPR/Cas9 delivery; bioengineered vectors; breast cancer treatment; synthetic biology in medicine; targeted gene therapy
    DOI:  https://doi.org/10.1080/15257770.2025.2500042
  10. Crit Rev Microbiol. 2025 May 06. 1-22
    From resistance to treatment: the ongoing struggle with Acinetobacter baumannii.
    Naji Naseef Pathoor, Vijetha Valsa, Pitchaipillai Sankar Ganesh, Rajesh Kanna Gopal.
      Acinetobacter baumannii (A. baumannii) has become a major hospital-acquired pathogen, well-known for its rapid development of resistance to multiple antibiotics. The rising incidence of antibiotic-resistant A. baumannii presents a significant global public health challenge. Gaining a deep understanding of the mechanisms behind this resistance is essential for creating effective treatment options. This comprehensive review explores the understanding of various antibiotic resistance mechanisms in A. baumannii. It covers intrinsic resistance, acquired resistance genes, efflux pumps, changes in outer membrane permeability, alterations in drug targets, biofilm formation, and horizontal gene transfer. Additionally, the review investigates the role of mobile genetic elements and the clinical implications of antibiotic resistance in A. baumannii infections. The insights provided may inform the development of new antimicrobial agents and the design of effective infection control strategies to curb the spread of multidrug-resistant (MDR) A. baumannii strains in healthcare environments. Unlike previous reviews, this study offers a more integrative perspective by also addressing the pathogen's environmental resilience, with particular emphasis on its resistance to desiccation and the formation of robust biofilms. It further evaluates both established and emerging therapeutic strategies, thereby expanding the current understanding of A. baumannii persistence and treatment.
    Keywords:  Acinetobacter baumannii; antibiotic resistance; biofilm; control measures
    DOI:  https://doi.org/10.1080/1040841X.2025.2497791
  11. Microb Drug Resist. 2025 May 09.
    Evaluating the Antibiofilm Effects of Antibiotics on Staphylococcus Species from Pediatric Hematology-Oncology Patients.
    Cansu Vatansever, Nilay Aksoy, Başak Adaklı Aksoy, Tunç Fışgın.
      Biofilms are microbial communities and occur on different medical devices such as catheters. The formation of bacterial biofilms on medical devices leads to indwelling medical device-related infections. Since biofilm bacteria are more resistant to antibiotics than planktonic bacteria, using these antibiotics in indwelling medical device-related infections causes recurrence of infections, treatment failure, and death. Minimum inhibitory concentration (MIC) is an important reference in treating acute infections caused by planktonic bacteria. However, MIC is ineffective in indwelling medical device-related infections caused by biofilm bacteria. The study aims to demonstrate the necessity and development of effective and standard methods such as minimum biofilm prevention concentration, minimum biofilm inhibitory concentration, and minimum biofilm eradication concentration in the case of indwelling medical device-related infection. The study was conducted with 10 isolates of Staphylococcus species from patients who developed infections in the Pediatric Hematology-Oncology Department at Medical Park Bahcelievler Hospital. According to the study results, even if planktonic bacteria are sensitive to antibiotics, they can become resistant to this antibiotic when they are in a biofilm (p < 0.05, Crosstab). Also, inhibiting the growth of planktonic bacteria does not prevent biofilm formation. The study additionally revealed that inhibiting and eradicating biofilm is more difficult than preventing biofilm formation (p < 0.05).
    Keywords:  Staphylococcus spp.; antibiotic resistance in biofilm; biofilm inhibition; biofilm prevention; indwelling medical device-related infections; pediatric hematology–oncology
    DOI:  https://doi.org/10.1089/mdr.2025.0006
  12. Wound Repair Regen. 2025 May-Jun;33(3):33(3): e70014
    A review on smart dressings with advanced features.
    Neda Akhavan-Kharazian, Hossein Izadi-Vasafi, Masih Tabashiri-Isfahani, Hossein Hatami-Boldaji.
      Wound care is a multifaceted and collaborative process, and chronic wounds can have significant repercussions on a patient's well-being and impose a financial burden on the healthcare industry. While traditional wound dressings can effectively facilitate healing, their limitations in addressing the intricacies of the wound healing process remain a formidable obstacle. Smart wound dressings have emerged as a promising solution to tackle this challenge, offering numerous advantages over conventional dressings, such as real-time monitoring of wound healing and enhanced wound care management. These advanced medical dressings incorporate microelectronic sensors that can monitor the wound environment and provide timely interventions for accelerated and comprehensive healing. Furthermore, advancements in drug delivery systems have enabled real-time monitoring, targeted therapy, and controlled release of medications. Smart wound dressings exhibit versatility, as they are available in various forms and can be utilised for treating different types of acute or chronic wounds. Ultimately, the development of innovative wound care technologies and treatments plays a vital role in addressing the complexities presented by wounds and enhancing patients' quality of life. This review sheds light on the diverse types of smart dressings and their distinctive features, emphasising their potential in advancing the field of wound care.
    Keywords:  drug delivery; sensor; smart wound dressing
    DOI:  https://doi.org/10.1111/wrr.70014
  13. Mol Nutr Food Res. 2025 May 04. e70090
    Development of a Korean Nutrition Model for In Silico Gut Microbiome Analyses Integrated With Nutrigenomics.
    Kyoung Su Kim, So-Yeon Yang, Hahyun Jeong, Minji Hong, Jihye Noh, Hong Koh, Dong-Woo Lee.
      The gut microbiome plays a crucial role in human health and disease, with diet serving as a critical determinant of microbial composition and metabolic function. However, most existing nutrition databases are Western-centric, lacking comprehensive dietary information for non-Western populations, including Koreans. This limitation hinders the accuracy of in silico gut microbiome analyses and microbiome-disease associations. We developed the Korean Nutrition Model (KNM) to enhance in silico microbiome analyses by incorporating detailed macronutrient and micronutrient compositions reflective of Korean dietary patterns. KNM was constructed using a decision algorithm that integrates data from the Ministry of Food and Drug Safety and FooDB. Comparative analysis with the European Nutrition Model revealed significant differences in carbohydrate and vitamin compositions, which in turn influenced microbial growth rates and metabolic fluxes in in silico simulations. We further evaluated gut microbiota differences between Korean and European cohorts, including healthy individuals and inflammatory bowel disease patients. Our findings demonstrate that using an appropriate, population-specific nutrition model significantly improves microbiome analyses, reducing the risk of false associations. This study underscores the importance of regionally tailored dietary models and provides a framework for enhancing global dietary models to facilitate precision nutrition and microbiome-based disease interventions.
    Keywords:  Korean Nutrition Model; diet; gut microbiome; in silico analyses; nutrigenomics
    DOI:  https://doi.org/10.1002/mnfr.70090
  14. Iran J Microbiol. 2025 Apr;17(2): 229-238
    Resistance profiles of Staphylococcus aureus isolates against frequently used antibiotics at private sector laboratories in Jordan.
    Rania Al-Groom, Ghina Al-Saraireh, Sultan Ayesh Mohammed Saghir, Mohd Sajjad Ahmad Khan, Areej M Almanaseer, Laila Alswalha, Wesal Alraei, Dalia Abu Al-Haijaa, Maha Hdaib, Anas Da'meh, Shereen Z Burjaq, Omar Al-Dmour, Fuad Alhawarat.
       Background and Objectives: Staphylococcus aureus (S. aureus) is one of the most important pathogens, responsible for a range of infections. This study aimed to assess resistance patterns in S. aureus isolates obtained from certain private-sector laboratories against commonly used antimicrobial agents.
    Materials and Methods: The process involved collecting various samples from several private laboratories and then identifying S. aureus isolates using biochemical characterization. The antibiotic susceptibility of these isolates was determined by disc diffusion method. Furthermore, Rt-PCR was employed to identify two genes namely the methicillin/oxacillin resistance genes (mecA), and (SCCmec).
    Results: The findings of the current study exhibited that females constituted a larger proportion of the participants (59.1%) compared to males (40.9%), with a mean participant age of 40.82 years. Gram-positive bacteria were more prevalent (71.3%) than Gram-negative bacteria (18.3%), with S. aureus being the most frequent isolate (60.9%). Urine samples represented the highest collected sample type (47.8%). Out of the 115 bacterial isolates, 85.2% exhibited multidrug resistance to antibiotics such as cefazolin, gentamicin, vancomycin, and ceftazidime. Clindamycin was the most effective antibiotic, with a sensitivity rate of 62.9%, followed by teicoplanin and meropenem, each with a sensitivity rate of 52.9%. Methicillin-resistant Staphylococcus aureus (MRSA) strains were susceptabile to vancomycin and teicoplanin. The methicillin/oxacillin resistant isolates showed significant association with mecA and SCCA genes.
    Conclusion: This study highlighted the multi-drug resistance in S. aureus isolates, stressing the need for stringent antibiotic stewardship, continuous surveillance, and further research into alternative treatments, including novel antibiotics and combination therapy, to combat resistant strains.
    Keywords:  MecA; Methicillin-resistant Staphylococcus aureus strains; Multidrug-resistant organisms; SCCmec; Staphylococcus aureus
    DOI:  https://doi.org/10.18502/ijm.v17i2.18382
  15. J Vis Exp. 2025 Apr 18.
    Cystic Fibrosis Aggregate Biofilm Model to Study Infection-relevant Gene Expression.
    Hollie J Leighton, Tegan M Hibbert, Grace I Ritchie, Daniel R Neill, Joanne L Fothergill.
      Standard pre-clinical testing methods for novel antimicrobial therapeutics used to treat chronic lung infections in people with cystic fibrosis do not reflect the environmental conditions of the hostile lung niche. Current reductionist testing conditions can lead to the progression of compounds along a preclinical pipeline without evidence of their activity under cystic fibrosis lung niche-appropriate conditions. Several approaches used to study traditional antimicrobials may not be suitable for antibiotic alternatives, including anti-virulence therapeutics like anti-quorum sensing agents and siderophore inhibitors. This protocol documents an aggregate biofilm model of Pseudomonas aeruginosa to compare resistance and infection-relevant gene expression in single-species and multi-species cultures (Staphylococcus aureus and Candida albicans), examining colony-forming unit (CFU) reductions and changes in gene expression, using algD as an exemplar. The model was optimized for small, static volumes of bacterial cultures to allow the study of novel compounds in the discovery phase of the drug development pipeline, where compound quantities may be limited. Single-species P. aeruginosa biofilms were formed in Synthetic Cystic Fibrosis Medium 2 (SCFM2) for 24 h before treatment with meropenem at different concentrations (1, 16, and 256 µg/mL) for a further 24 h. Polymicrobial biofilms were established by growing Staphylococcus aureus and Candida albicans together in SCFM2, then inoculating with P. aeruginosa for an additional 24 h and treating with meropenem. The lack of a direct connection between compound efficacy measures in pre-clinical testing and clinical trial results has cast doubt on the applicability of current laboratory screening tools. This model allows us to understand the impact of relevant factors on P. aeruginosa gene expression, including genes contributing to resistance and virulence, thereby bridging this gap.
    DOI:  https://doi.org/10.3791/67477
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