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. 2023 Aug 28;12(9):1090.
doi: 10.3390/pathogens12091090.

Evaluation of a Tetracycline-Resistant E. coli Enumeration Method for Correctly Classifying E. coli in Environmental Waters in Kentucky, USA

Callie Boggs  1 , Kidus Shiferawe  1 , Eckhardt Karsten  2 , Jayden Hamlet  3 , S Travis Altheide  4 , Jason W Marion  1   5
Affiliations

Evaluation of a Tetracycline-Resistant E. coli Enumeration Method for Correctly Classifying E. coli in Environmental Waters in Kentucky, USA

Callie Boggs et al. Pathogens. .

Abstract

The global concern over antimicrobial resistance (AMR) and its impact on human health is evident, with approximately 4.95 million annual deaths attributed to antibiotic resistance. Regions with inadequate water, sanitation, and hygiene face challenges in responding to AMR threats. Enteric bacteria, particularly E. coli, are common agents linked to AMR-related deaths (23% of cases). Culture-based methods for detecting tetracycline-resistant E. coli may be of practical value for AMR monitoring in limited resource environments. This study evaluated the ColiGlow™ method with tetracycline for classifying tetracycline-resistant E. coli. A total of 61 surface water samples from Kentucky, USA (2020-2022), provided 61 presumed E. coli isolates, of which 28 isolates were obtained from tetracycline-treated media. Species identification and tetracycline resistance evaluation were performed. It was found that 82% of isolates were E. coli, and 18% were other species; 97% were identified as E. coli when using the API20E identification system. The MicroScan system yielded Enterobacter cloacae false positives in 20% of isolates. Adding tetracycline to ColiGlow increased the odds of isolating tetracycline-resistant E. coli 18-fold. Tetracycline-treated samples yielded 100% tetracycline-resistant E. coli when the total E. coli densities were within the enumeration range of the method. ColiGlow with tetracycline shows promise for monitoring tetracycline-resistant E. coli in natural waters and potentially aiding AMR surveillance in resource-limited settings among other environments.

Keywords: Enterobacter; Enterobacter cloacae; Escherichia coli; antibiotic resistance; antimicrobial resistance; culturable; monitoring; one health; tetracycline resistance; water quality.

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Conflict of interest statement

C.B., K.S., E.K., J.H. and T.S.A. declare no conflict of interest. J.W.M. declares being the inventor on an international PCT application and pending U.S. patent application by Eastern Kentucky University for a method for the detection of E. coli and antibiotic-resistant bacteria in water. J.W.M. declares being the principal for Eastern Scientific LLC, which is licensed by Eastern Kentucky University to commercialize the method. Eastern Scientific LLC has a pending U.S. trademark application for ColiGlow™. The funders of the research had no role in the design of the study, in the collection, analyses, interpretation of data, writing of the manuscript, or in the decision to publish the results. The content provided here represents the original findings of the authors and does not necessarily represent the views or interpretations of Eastern Kentucky University, Miami University of Ohio, Stockton University, or the National Science Foundation.

Figures

Figure 1
Figure 1
The results of one water sample evaluated for E. coli density by the ColiGlow method without tetracycline (left), and the ColiGlow method containing tetracycline (right) whereby the fluorescing wells under longwave ultraviolet light are indicative of presumed E. coli growth.
See this image and copyright information in PMC

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