2020: Selecting the sites for monitoring and investigations. Experiments set-up and dissemination of results.

Sites from lotic and soil ecosystems with low anthropogenic impact and presumptive contrasting biodiversity were identified by all partners of the consortium. A common approach in sampling methodology (SOP - Standard Operational Procedure) of biofilm samples/sediments from freshwater and of soil was developed. Sampling for the identification of sites with low or high microbial diversity for further investigation in the other activities of the project was performed.

In the case of the Romanian partner, areas with varying anthropogenic impacts have been identified in the Someșul Mic Basin, followed by the selection of points with presumptive contrasting microbial biodiversity and biological sampling campaigns (biofilm and sediments). For soil sampling, brown soils were selected from a varied geologic substrate, together with agricultural soil. The SOP were established in the project consortium, as a result of the effort of the various working groups involved. The document details the methodology for sampling heterotrophic biofilms and sediment in aquatic habitats and soil samples, their transport to the laboratory and the nucleic acid isolation protocol.

Prototypes of artificial exposure units were developed and tested, for subsequent use in experiments investigating the barrier effect of microbial biodiversity in water and soil environments. The Romanian partner was part of the working group focused on developing the experimental model for capturing the contrasting microbial diversity in lotic ecosystems and building of the most suitable prototype. In addition, a laboratory was set up to allow long-term microcosm experiments at controlled temperature for both aquatic and soil samples and a large volume filtration system was purchased to filter off-site waters to be used in project experiments.

The main goal of the ANTIVERSA project is to investigate the possible barrier effect imposed by microbial communities on the accumulation and spread of antibiotic resistance in the environment. This phase of the project was focused on a better understanding of the complex phenomenon of anthropogenic impacts by wastewater treatment plants through the spread of antibiotic resistant/multi-resistant pathogens in the environment.

The preliminary results, the experimental plans and the development of prototypes have been assessed continuously for optimization. Discussions were held with representatives of various stakeholders regarding the ANTIVERSA outputs and how can they be involved in the project implementation phases.