Global efforts emphasize the need to incorporate human, animal, and environmental components into coordinated and standardized surveillance programs. This requires microbiological methods be standardized, including using standardized positive and negative bacterial control strains. There is a need to provide bacterial control strains that are sourced from and relevant for environmental surveillance efforts.· Two control strains are available in support of environmental antibiotic resistance surveillance efforts.· Escherichia coli ARS-C301 is extended-spectrum β-lactamase positive and tetracycline resistant.· E. coli ARS-C101 is cefotaxime and tetracycline susceptible and serves as a negative control.· Strains are DNA sequenced, and deposited in publicly available culture collections.Strain Availability:ARS-C101 (negative control): WGS is available from NCBI, accession number PRJNA1003888, BioSample B-65681; Isolate is available from the ARS Culture Collection https://nrrl.ncaur.usda.gov/ (Accession number: B-65681); and from ATCC (isolate ID BAA-3340).ARS-C301 (positive control): WGS is available from NCBI, accession number PRJNA1003888, BioSample SAMN36910816; Isolate is available from the ARS Culture Collection https://nrrl.ncaur.usda.gov/ (Accession number: B-65682); from ATCC (isolate ID BAA-3341); and from Microbiologics as a quantitative pellet, via a custom order (contact industrial microbiology QC products regional sales manager by email requesting a quote for ARS-C301 https://www.microbiologics.com/contact-us).