Antibiotic resistance (AR) infections are a pressing environmental health issue. Many AR infections in humans come from exposure to soil, water, and air. New methods are thus needed to measure the risk associated with environmental AR. This project seeks new techniques to identify genetic markers for AR microbes in different environmental settings. This information will be used to track the transmission of these markers through the environment. The project will educate students in environmental engineering and interdisciplinary problem solving to tackle complex problems like AR transmission. Research opportunities will also be created for Puerto Rican and Hispanic students to broaden understanding and participation in STEM. Results will be shared with the public in English and Spanish to educate diverse communities on the link between the environment and human health. Knowledge gained from this project will help transform environmental biotechnology to address environmental AR for the protection of human and ecological health.<br/><br/>The link between human, animal, and environmental health (One Health) is exemplified by one of our most pressing public health issues: antibiotic resistance. Environmental hot spots of antibiotic resistance have been identified using techniques that do not identify the genetic context of antibiotic resistant genes (ARGs). Failure to understand the microbial ecology and mechanisms driving the proliferation of ARGs in the environment limits our ability to characterize the hazard posted by these genes and prevents the development of engineering solutions to limit the spread of antibiotic resistant infectious disease. To address these issues, field surveys and datamining are proposed to: 1) provide guidance on best practices for linking functional genes to putative host cells in metagenomic data from mixed microbial communities; 2) characterize the diversity of the ARG carrying host cells; and 3) determine the prevalence of waterborne pathogens carrying an ARGs. Samples from these studies will help define the relative abundance of select ARGs across air, water, and biofilms. A laboratory experiment will be performed to determine the relative contribution of host growth versus horizontal gene transfer (HGT) under selective pressure. If a minimum concentration to promote HGT can be observed, the genetic context of environmental ARG will be defined under selecting conditions. By studying host cells, horizontal gene transfers, and genetic context as a function of water quality, this research will contribute fundamental knowledge on the factors controlling HGT in water. This information can be used towards design of engineered treatment systems and is of critical importance for understanding the transfer of resistance and understanding conditions that promote beneficial and limiting transfers of functional genes of importance. The data generated will help fulfill a critical data gap for quantitative microbial risk assessment for ARGs. The project will provide Broader Impacts by improving curriculum, promoting network building, and facilitating interdisciplinary problem solving through an interdisciplinary course. It will also build bridges to education in environmental engineering with Hispanic students and communities. Results of the research will be shared at the Rutgers Day public open house in English and Spanish.<br/><br/>This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.