Development and in vivo validation of a novel triggered release antibiofilm coating for orthopaedic implants and vascular grafts. (TRA-COAT)

One of the most challenging problems related to implanted medical devices is the high prevalence of microbial infections. Previous studies showed that implant-related infections are often caused by biofilm forming microbes. In contrast to infections caused by free-floating microbes, biofilms generally increase the tolerance to antimicrobials and increase the risk for antimicrobial resistance development.

The TRA-COAT project aims to address the incidence of implant infections by developing a novel, resistance-proof antimicrobial coating with innovative functionalities, including antibiofilm activity and triggered on-demand release of classic antimicrobials. The focus will be on orthopaedic trauma implants and vascular grafts, but can later be extended to other implants.

The coating will combine a covalently bonded biofilm inhibitor and a reversibly bonded classic antimicrobial. The action of the inhibitor will reduce the slimy matrix production during biofilm formation which will allow the immune system to better remove the microorganisms. If biofilm should still remain, growth of the bacteria on the surface will locally trigger release of the antimicrobial, killing the pathogens.
The coating is designed to be resistance-proof. Indeed, the biofilm inhibitors are intrinsically robust to resistance and will enhance the effectivity of the classic antimicrobial. The local and time-restricted release of the antimicrobial will reduce the risk of resistance development and benefit the patient.

Project partners

  • Hans Steenackers, Katholieke Universiteit Leuven, Belgium (Coordinator)
  • Stephan Zeiter, AO research institute, Switzerland
  • Annette Moter, Charité – Universitaetsmedizin Berlin, Germany


Disrupting drug resistance by exploiting collateral sensitivity to design novel therapeutic strategies for C. auris and C. glabrata (CycleDrug)

Multidrug resistance is a major problem in Candida auris, the first pathogenic fungus officially considered an urgent antimicrobial resistance threat by the CDC, and in Candida glabrata, which accounts for 20-40% of all systemic Candida infections. Antifungal resistance often leads to treatment failure, which significantly reduces survival rates of lethal candidiasis. Meanwhile, the antifungal drug market comprises only four classes.

By evolving C. auris and C. glabrata in different drugs and mapping their responses to other drugs, we have discovered collateral sensitivity (CS) and cross resistance (XR). CS is the process in which the acquisition of drug resistance towards one drug, confers an increased sensitivity towards another drug. Conversely, XR confers reduced susceptibility to more than one drug upon exposure to one drug. Information regarding the evolutionary tendencies of pathogenic fungi can be leveraged to improve therapeutic approaches for treating fungal infections. Both CS and XR have been studied extensively in tumors and in bacteria but remain unexplored in fungi. In this study, we will explore novel treatment schemes that have the potential to prevent the development of antifungal drug resistance in MDR species of most concern: C. auris and C. glabrata.

Project partners

  • Patrick Van Dijck, Katholieke Universiteit Leuven, Belgium (Coordinator)
  • Katrien Lagrou, University Hospitals Leuven, Belgium
  • Johan Maertens, University Hospitals Leuven, Belgium
  • Micha Fridman, Tel Aviv University, Israel
  • Juan Antonio Gabaldon Estevan, Institute for Research in Biomedicine, Spain
  • Berman Judith, Tel Aviv University, Israel


New Inhalation Formulations for Improved Delivery of Antimicrobials (APRINHA)

Our body’s branching airway system that delivers oxygen deep into our lungs presents an effective barrier to protect our lungs from contaminants in the air, such as dust, pollen, and pollutants, but also bacteria and viruses.

The same barrier that protects us from inhaling unwanted matter also presents a considerable hurdle in using the human airways as an effective route of drug delivery. A variety of respiratory diseases benefit from inhaling a drug to route its active ingredient directly to its desired location in the human lung, rather than taking a detour through the stomach or blood circulation. Respiratory infections by bacteria, also known as bacterial pneumonia, can turn into life-threating conditions that could more easily be cured with powerful antibiotic inhalations. A complicating factor in the development of inhaled antibiotics is the fact that large amounts of antibiotics need to be applied, of which only a fraction end up at the actual site of infection.

Attempts to improve existing inhalation technologies have seen some progress in recent years. The APRINHA project aims to leverage on some of these achievements to study and compare a variety of novel formulation technologies using the new antibiotic apramycin as a promising case study. Apramycin is expected to be perfectly suited for such studies because it belongs to a drug class that has already been shown to be suited for inhalation. The project aims to develop an antibiotic inhalation with high penetration, deposition, retention, and efficacy of drug, so as to more effectively cure pneumonia patients.

Project partners

  • Sven Hobbie, University of Zurich, Switzerland (Coordinator)
  • Dorothee Winterberg, Fraunhofer Institute for Toxicology and Experimental Medicine, Germany
  • Iraida Loinaz, CIDETEC, Spain
  • Frédéric Tewes, INSERM U1070, France
  • Edgars Liepins, Latvian Institute of Organic Synthesis, Latvia
  • Anna Fureby, RISE Reaserch Institute of Sweden AB, Sweden
  • Per Gerde, Inhalation Sciences Sweden AB, Sweden


Design and implementation of effective cOmbination of Phages and Antibiotics for improved TheRApy protocols against KLEbsiella pneumoniae (KLEOPATRA)

The multidrug-resistant bacteria Klebsiella pneumoniae represents a critically emerging pathogen that confounds treatment in a clinical setting. This is due to their pan-resistance to antibiotics and specific biological traits, including a protective capsule, that makes it insensitive to the host immune system.

Bacterial viruses, also called (bacterio)phages are natural predators of bacteria. Our previous research has identified phages that overcome the capsule barrier by specific enzymes and resensitize bacteria to the innate immune system as well as to antibiotic treatment.

In this project, we introduce the concept of “K-sensitization” which uses these phages and their enzymes (capsule depolymerases) to strip the protective capsules from prevalent K. pneumoniae strains and provide a proof-of-concept of how phages/depolymerases act synergistically to antibiotic treatment. Besides the development of a protocol for compassionate phage therapy against these encapsulated pathogens, we will comprehensively analyze the prevalent K. pneumoniae circulating in human & animal reservoirs, as well as environmental ecosystems. This in turn will lead to the establishment of tailored phage banks & associated diagnostics tools, supported by computational models to ensure rationally designed phage therapy cocktails for antibiotic/phage/enzyme combination treatments.
In this manner, the KLEOPATRA consortium aims to contribute to improving the treatment of this critical WHO priority 1 pathogen within a ‘One Health’ setting.

Project partners

  • Zuzanna Drulis-Kawa, University of Wroclaw, Poland (Coordinator)
  • Rob Lavigne, KU Leuven, Belgium
  • Regis Tournebize, Sorbonne Université, France
  • Joachim J. Bugert, Bundeswehr Institute of Microbiology, Germany
  • Jens Andre Hammerl, German Federal Institute For Risk Assessment, Germany
  • Ronen Hazan, Hebrew University of Jerusalem, Israel
  • Kilian Vogele, INVITRIS SME, Germany


Antimicrobial Stewardship in Hospitals, Resistance Selection and Transfer in a One Health Context (STRESST)

The transfer of antibiotic residues and antibiotic resistant bacteria into the environment and subsequently into animal drinking water may have an effect on the transmission of resistant bacteria and their resistance genes back into the human population.

This holistic One Health view of antibiotic resistance is at the heart of our project. We want to determine if hospital wide antimicrobial stewardship implementation will reduce antibiotics and antibiotic resistant bacteria from entering the environment and if the reduction of antibiotic concentrations will lower the transfer of resistance genes within and between bacteria in the environment and in animals. We will show that hospital wastewater is a hotspot for selection of resistance and pave the way for future, targeted interventions aimed at reducing the amounts of antibiotics released into the environment even further.

Project partners

  • Adam Roberts, Liverpool School of Tropical Medicine, United Kingdom (Coordinator)
  • Andrew Singer, UK Centre for Ecology and Hydrology, United Kingdom
  • Nina Langeland, University of Bergen, Norway
  • Michael Brouwer, Wageningen Bioveterinary Research, Netherlands


Specific Targeting of Antimicrobial Resistant Strains in situ using Targeted-Antibacterial-Plasmids (STARS-TAP)

The global spread of antimicrobial resistance (AMR) among pathogenic bacteria is recognized as one of the biggest concerns in public health and a research priority in microbiology.

Drug-resistance increases exponentially for certain bacterial organisms and is becoming the main threat to human health worldwide. As a consequence, national and international authorities have emphasized the need to taking a broad, coordinated approach to develop new antimicrobial strategies to fight against drug-resistant bacteria across multiple sectors such as human health and animal health, agriculture and environment1 (i.e. ‘One health’ major challenge).

The STARS-TAP research program aims at developing an innovative non-antibiotic antibacterial methodology to specifically target AMR strains from natural bacterial communities in several ecosystems in situ. The proposed methodology is based on Targeted-Antibacterial-Plasmids (TAPs) that use DNA conjugation to deliver CRISPR/Cas systems exerting a strain-specific antibacterial activity. If successful, our research would represent a real breakthrough for clinical and environmental microbiology, and open new options for the elimination of AMR strains from various ecosystems. This unexplored and versatile strategy complementary to antibiotic treatments holds the potential to be used for preventive decolonization purposes, or even in agriculture to tackle AMR prevalence in amended soils.

Project partners

  • Christian Lesterlin, Centre National de la Recherche Scientifique, France (Coordinator)
  • Pierre Bogaerts, UCL – Université Catholique de Louvain, Belgium
  • Gregory Jubelin, French National Research Institute for Agriculture, Food and Environment, France
  • Anna Marzec-Grzadziel, Institute of Soil Science and Plant Cultivation – State Research Institute, Poland
  • William Couet, French National Institute of Health and Medical Research, France


Strengthening implementation of National Action Plans through a One Health AMR full economic costing exercise (SNAP ONE)

Antibiotic resistance costs lives and money. Yet if we don’t have a good grasp of the numbers we will never know where it lies in terms of our other national priorities.

In Africa we have especially little evidence on how people, animals, and the environment are affected by it. So those who make decisions do not see it at a particular problem and, as a consequence, those who hold the purse strings in government do not fund the necessary efforts to combat antibiotic resistance.

This project will estimate the full economic burden imposed by antibiotic resistance (epidemiological and financial) in Malawi and Zambia in order to fill this evidence gap. Findings from this work should help enhance political will to take greater action. It will also allow more informed decisions to be made about how best to tackle antibiotic resistance locally by calculating the important cost-related numbers that allow different strategies to be compared. Crucially this project will bring together the key people needed to make real and further progress on this issue each of these two countries. And, if successful, Malawi and Zambia will be setting the example for how to combat antibiotic resistance in other parts of Africa.

Project partners

  • Chantal Morel, University Hospital Bonn, Germany (Coordinator)
  • Finola Leonard, University College Dublin, Ireland
  • Chisomo Msefula, University of Malawi, College of Medicine, Malawi
  • Luigia Scudeller, Azienda Ospedaliero-Universitaria di Bologna, Italy
  • Lloyd Matowe, Eden University, Zambia
  • Herman Goosens, University of Antwerp, Belgium


Selecting Efficient Farm-level Antimicrobial Stewardship Interventions from a one health perspective (SEFASI)

Antimicrobial resistance (AMR) links together people, plants, animals and their environments under the One Health umbrella. In this work we will similarly link interventions aimed at AMR by considering their impact not only in terms of impact on hospitals, communities or farmers, but across all of these groups.

This is key to informing optimal intervention selection by governments in tackling AMR in the future. Our research will combine statistical analysis, mathematical simulations and economic-impact models within a single intervention assessment framework. We will bring together an interdisciplinary team of economists, mathematical modellers and veterinary scientists to apply this modelling framework to three country cases studies: England, Senegal and Denmark. All three countries are global leaders in terms of AMR data collection and intervention, providing ideal settings for intervention assessment. Our outcome will be a ranking of farm-level interventions for policymakers to assess their impact from a One Health perspective, and an insight into where more data in the future would be most beneficial, in terms of reducing uncertainty in such economic evaluations of interventions.

Project partners

  • Gwenan Knight, London School of Hygiene and Tropical Medicine, United Kingdom (Coordinator)
  • Michel Dione, International Livestock Research Institute, Senegal
  • Ana Mateus, Royal Veterinary College, United Kingdom
  • Nichola Naylor, Public Health England, United Kingdom
  • Dagim Belay, University of Copenhagen, Denmark


Phage Therapy to Reduce AMR Enterobacteria Spread from a One Health Perspective (Phage-Stop-AMR)

The spread of multi-drug resistant (MDR) bacteria in food-producing animals including broilers is a global public health concern.

Controlling growth of MDR bacteria and limiting the transmission of antimicrobial resistance genes in broilers could be an effective mitigation strategy. To counteract the spread of MDR bacteria among zoonotic pathogens in food-producing animals and reduce the risk of their transmission to humans or the environment, antibiotic use in animal husbandry has to be reduced. Bacteriophage therapy is increasingly accepted as an environmentally-friendly antimicrobial intervention strategy, effective at specifically targeting bacterial pathogens, to prevent the transmission of resistant bacteria from foods to humans and vice versa.

We use MDR Salmonella and E. coli in broilers as a model and will first select the most efficient phage combinations to specifically reduce these bacteria and MDR plasmids in broilers. Using laboratory, an experimental chicken gut model and farm-level experiments, we will then establish the efficacy of phage formulations as feed additives within a commercial farming context to reduce bacterial numbers and progressively reduce MDR plasmid carriage in broilers. We will test the effect of phage therapy on intestinal parameters of the treated broilers and also on the broiler intestinal microbiome and resistome composition. We will investigate the transmission of AMR plasmids between different enterobacteria in the broiler gut and improve on-site detection of MDR foodborne pathogens as an early warning system at farm level.

Project partners

  • Ulrich Dobrindt, Universität Münster, Germany (Coordinator)
  • Clara Marín-Orenga, Universidad Cardenal Herrera – CEU, Spain
  • Muna Anjum, Animal and Plant Health Agency, United Kingdom
  • Raul Fernandez Lopez, Universidad de Cantabria, Spain
  • Danish Malik, Loughborough University, United Kingdom
  • Annamária Szmolka, Veterinary Medical Research Institute, Hungary
  • Eliora Ron, Tel Aviv University, Israel


Phage treatment and wetland technology as intervention strategy to prevent dissemination of antibiotic resistance in surface waters (PhageLand)

PhageLand is aimed to develop a novel intervention strategy combining the low-cost and eco-friendly capacity of constructed wetlands with the specificity of bacteriophages (i.e., viruses killing bacteria) to prevent the dissemination of antibiotic resistance from wastewater into surface waters.

PhageLand will investigate the prevalence of antibiotic resistant bacterial pathogens (ARB) in low-middle income countries (LMICs) in Eastern Europe, which will be then used as targets for the development of a dedicated phage-based treatment for their specific removal from communal wastes.

In parallel, PhageLand will assess: a) the purification capacity of two reference, full-scale constructed wetlands operating in Spain and Moldova in the removal of antibiotic residues, ARB and antibiotic resistance genes; and b) the potential risk associated with the dissemination of these biological pollutants within indigenous bacterial communities and among animals inhabiting constructed wetlands.

Finally, PhageLand will develop a pilot plant to scale-up the phage-wetland combined technology to assess its performance under real environmental conditions. This proof-of-concept will be used to demonstrate the efficacy of this nature-based technology for the removal of multidrug-resistant pathogens from communal wastes and to encourage stakeholders for its implementation in wastewater treatment to prevent the dissemination of antimicrobial resistance. The PhageLand technology will be particularly useful in LMICs, where costly and power-demanding treatment plants are difficult to set up.

Project partners

  • Carles Borrego, Catalan Institute for Water Research, Spain (Coordinator)
  • Lukasz Dziewit, University of Warsaw, Poland
  • Malgorzata Grzesiuk-Bieniek, Warsaw University of Life Sciences, Poland
  • Rob Lavigne, KU Leuven, Belgium
  • Evelien Adriaenssens, Quadram Institute Bioscience, United Kingdom
  • David Weissbrodt, Delft University of Technology, Netherlands
  • Alina Ferdohleb, Nicolae Testemitanu State University of Medicine and Pharmacy, Moldova