The alarming rise in resistance to antibiotics is now widely accepted as being one of the most serious public health crises we face today. Recent studies indicate that the natural environment is a significant contributor to the emergence and transmission of resistant pathogens. Resistance is selected in environments polluted with antibiotic residues, heavy metals and other chemicals of emerging concern. The River Almond catchment area has all the ingredients of an antibiotic resistance transmission and selection ''hotspot''. Including nine waste-water treatment plants, numerous combined sewer overflows and septic tanks, farm animal waste, mining discharge and landfill sites. This projects will quantify the levels of antibiotic resistant bacteria and genes in a river catchment, evaluate potential sources of antibiotic resistance pollution and assess links between potential sources, prevalence of resistance and concentration of chemicals which select resistance. Such surveillance is key to inform public health interventions and the baseline data gathered in this project will inform suitable intervention targets. Aim To determine the abundance of antibiotic resistance bacteria (ARB) and genes (ARG) and investigate the links with antibiotic resistance selective chemicals along a river impacted by multiple pollution sources in order to provide a baseline for future risk assessment and intervention strategies. Objectives 1. Use culture dependent and qPCR to detect and quantify ARB and ARG along the length of the River Almond. 2. Identify the major sources of antibiotic resistance pollution in the River Almond, focusing on point sources (WWTP, combined Sewer Overflow, mining discharge) and diffuses sources (agricultural sites) using GIS data and Microbial Source Tracking. 3. Determine the potential relationship between the abundance of ARB and ARG and the concentration of antibiotic resistance selective chemicals. 4. Using metataxonomic and WGS investigate whether ''naturalized'' (environmental) E. coli, as distinct from enteric E. coli, can be used as indicators of environmental selection of resistance.