Transnational Access Project: Annular Flume studies to test the effect on antibiotic resistant genes and use of CRISPR-Cas in E. coli from sediments affected by sewage pollution
Institutions involved
- University of Sheffield (SFD), United Kingdom
- Universitat Politècnica of Catalunya, Spain
- Universitat Politècnica de Catalunya-BarcelonaTech, Spain
Duration
14/03/2023-19/06/2023
About the project
The annular flume facility at the University of Sheffield was used for a Transnational Access led by Universitat Politècnica of Catalunya (UPC), investigating the effect of hydrological events on the presence of antibiotic resistance bacteria in rivers affected by faecal pollution. Whilst in the domain of urban drainage, this project has a heavy microbiological and genetic focus.
This project had a 2-phase experiment, with a part involving the annular flume (a circular flume creating an “infinite” flow for the study of sediment in moving water) and a part done in a bio-laboratory. The annular flume was involved determining if antibiotic resistant bacteria could be transferred between biofilms attached to sediments and river water. Biofilms were grown in the annular flume using water from a local river, and then the presence of the antibiotic-resistant bacteria was characterised in both the water and the sediment at varying levels of shear stress. The bio-laboratory tests involved the evaluation of CRISPR-Cas, a technique for editing genetic material, as a tool to control antimicrobial resistance in freshwater environments. Molecular biology experiments are very expensive, much more expensive than e.g. COD test kits. Whilst CRISPR-Cas is one of the most efficient and cost-effective gene editing tools, it is still very expensive when compared to basic biochemical testing.
Main findings
- Rivers that are deemed to be clean are, in truth, contaminated with a lot of disease-causing bacteria and antibiotic-resistant bacteria. Samples were not taken from a site that was anticipated to have high contamination levels as it was a recreational site used as a nature reserve for salmon. Stakeholders need to know that the rivers are contaminated so that a strategy to manage the pollution can be developed.
- The studies conducted in the annular flume provided valuable insights into the environmental and health impacts of antibiotic-resistant bacteria spreading in aquatic environments due to hydrological disturbances and showed proof of concept.
- The research generated crucial data for developing a CRISPR/Cas9-based strategy to effectively target and cleave the CTX-M-1 gene, with the goal of restoring antibiotic susceptibility in ESBL-producing bacteria, which is showing promise.
- More time and funding are needed to do repeats of this study to generate more robust conclusions and there are already ideas for the next steps if the opportunity arises. To continue the work on this project, partners are considering applying for further funding, likely at the European level.
Successful outcomes:
- This technology could in future be used to prevent the spread of antibiotic resistance spreading in the environment, which is an existing type of pollution with significant negative implications to health. As time goes on, this technology becomes cheaper and, thus, more accessible.
