Location: Agroecosystem Management Research
Project Number: 3042-12630-004-005-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Jun 16, 2020
End Date: Dec 16, 2022
Objective:
The overarching hypothesis is that non-MDR enterococci naturally present in manure and manure-impacted soils and waters use CRISPR-Cas to impede the spread of DNA in these environments. The first objective of this work is to use DNA sequencing to analyze the CRISPR memories of enterococci resident in manure and manure-impacted soil and water to determine what phage and plasmids are they encountering and acquiring memories against. The second objective experimentally assesses whether CC is an effective barrier to plasmid conjugation in manure microcosms.
Approach:
CRISPR memory of ARGs in manure-impacted habitats will be determined by targeted sequencing of Enterococcus field isolates. The Cooperator will provide the sequencing and analysis. ARS and ARs collaborators will provide the bacterial isolates. (O2) Tge Cooperator's in vivo mouse gut experiments will be replicated in microcosms simulating conditions in manure storage lagoons to determine if a functioning Enterococcus C-C also serves as a barrier to HGT in environmental manure storage structures. The Cooperator will use her Enterococcus isolates constructed for these analysis, and perform the studies in manure microcosms, with manure and microcosm design provided by ARS.
