National Science Centre, SONATA 19, “Is there an escape from antibiotic-resistant ESKAPE pathogens present in wastewater – the significance of last-resort antibiotics and the latest therapeutic options” July 10, 2024 – July 9, 2027

Project Manager: Katarzyna Natalia Grudlewska-Buda, PhD

Funding: PLN 305,280.00

Project Description: The widespread use of antibiotics, both in human medicine and in agriculture and animal husbandry, has led to an increase in the occurrence of microbial resistance. The “One Health” concept, promoted by the World Health Organization, emphasizes the need for close cooperation between human health, animal health, and the state of the environment. Wastewater treatment plants create excellent conditions for the spread of resistant bacteria in the environment through discharged wastewater, sewage sludge used for fertilization purposes, and in the form of generated bioaerosol. A particularly dangerous phenomenon is the emerging resistance to new drugs and so-called “last-resort” antibiotics used in cases of infections with multidrug-resistant strains. The occurrence of such microorganisms can vary depending on the type of wastewater and the season. For many years, the assessment of antibiotic resistance focused on examining indicator bacteria. However, there is still a lack of information on the spread of resistant bacteria, not only to well-known and commonly used antimicrobials, but also to last-resort antibiotics and those classified as new therapeutic options. For this reason, the aim of this project is to assess the frequency of ESKAPE group strains (Enterococcus, Staphylococcus, Klebsiella, Acinetobacter, Pseudomonas, Enterobacter) resistant to last-resort drugs and new therapeutic options isolated from various types of wastewater, including hospital wastewater. Furthermore, the project will enable the characterization of the genetic basis of detected resistance in the studied strain population. The project will involve collecting samples of screenings, raw sewage, primary sludge, treated sludge for agricultural use, and treated wastewater from three mechanical-biological wastewater treatment plants differing in terms of population equivalent (PE). Studies will be conducted using both traditional microbial culture methods and modern genome analysis techniques. Additionally, the project plans to detect antibiotics in wastewater, which will further expand the number of obtained data and help achieve the project’s objectives. The following tests will be performed: • detection of last-resort antibiotics and new therapeutic options in wastewater • assessment of the occurrence of ESKAPE pathogens in wastewater and sewage sludge • detection of ESKAPE bacteria with antibiotic resistance mechanisms • description of the susceptibility of isolated strains to antibiotics, especially last-resort antibiotics and new therapeutic options • identification of detected antimicrobial resistance genes and mobile genetic elements responsible for resistance to last-resort antibiotics and new therapeutic options. The basic scientific research planned in the project will provide new knowledge on the occurrence of antibiotic-resistant bacteria of greatest clinical importance in wastewater and will also enable the determination of their resistance to last-resort drugs and new therapeutic options. Genome analysis will enable the detection of genes conditioning this resistance. The project’s implementation will allow for an analysis of the consistency between pheno- and genotypic research results and will fill gaps in the current state of knowledge on the genetic determinants conditioning bacterial resistance. During the project’s implementation, it will be particularly important to determine whether the studied ESKAPE group bacteria spread and persist in wastewater at various stages of its treatment. Considering the undeniable health threat posed by antibiotic resistance and its spread in the environment, the current project aligns with the contemporary trend of research related to understanding the co-occurrence of antibiotic-resistant bacteria in different environments.