Evaluation of a modified IDEXX method for antimicrobial resistance monitoring of extended Beta-lactamases-producing Escherichia coli in impacted waters near the U.S.-Mexico border

Abstract

As part of a One Health approach, the World Health Organization (WHO) has deemed extended beta-lactamases-producing Escherichia coli (ESBL-Ec) as an appropriate proxy for antimicrobial resistance (AMR) in human, animal, and environmental samples. Traditional methods for ESBL-Ec quantification involve a labor-intensive process of membrane filtration, culturing in the presence and absence of antibiotics, and colony confirmation. The emerging modified IDEXX method utilizes IDEXX Colilert-18 test kits, recognized by the USEPA for the enumeration of total coliforms and E. coli in water samples, modified with cefotaxime for measurement of ESBL-Ec in environmental samples. However, this method has yet to be validated for ocean or sewage-contaminated water and has not been compared against the plate-based method with mTEC for surface water. In this study, ESBL-Ec in ocean and river waters of the Tijuana River Estuary were analyzed by three methods: membrane filtration using mTEC plates (as outlined in USEPA Method 1603), membrane filtration using TBX plates (as outlined in the WHO Tricycle Protocol), and Colilert-18 spiked with cefotaxime (Hornsby et al., 2023). Levels of ESBL-Ec were elevated in the Tijuana River Estuary and nearby ocean samples, as high as 2.2 × 10⁶ CFU/100 mL. The modified IDEXX method correlated with membrane filtration methods using selective mTEC (r = 0.967, p < 0.001, n = 14) and TBX (r = 0.95, p < 0.001, n = 14) agars. These results indicate that the modified IDEXX method can be used as a more accessible alternative to the traditional culturing methods as a screening tool for antibiotic resistance in urban aquatic environments. Advantages of the IDEXX-based method including portability, lower Biosafety Level requirements, fewer dilutions to stay within the dynamic range, greater ease of maintaining sterility during analysis, and less required staff training are discussed. Future studies into the validity of the modified IDEXX method compared to qPCR and metagenomic sequencing are needed.