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Updated: Jun 26, 2026

Isolation and Identification of Waterborne Antibiotic-Resistant Bacteria and Molecular Characterization of their Antibiotic Resistance Genes
Published on: March 3, 2023
Bacteria isolated from sewage influent resistant to ciprofloxacin, chloramphenicol and tetracycline
Sam R Zwenger1, Eric T Gillock
1School of Biological Sciences, University of Northern Colorado, Greeley, Colorado, USA.
Abstract:
This study assessed the presence of antibiotic-resistant bacteria in sewage influent. Resistance was measured by determining the lowest concentration of antibiotic, in micrograms per milliliter (microg mL(- 1)). To determine the minimum inhibitory concentration (MIC), which is used in diagnostic laboratories, we used the Etest, a plastic strip containing an antibiotic concentration gradient. In total, we sampled five sewage treatment plants of various sizes in Kansas and isolated bacteria resistant to three broad-spectrum antibiotics; ciprofloxacin (1-cyclopropyl-6-fluoro-4-oxo-7-piperazin-1-yl-quinoline-3-carboxylic acid), chloramphenicol 2,2-dichlor-N-[(aR, bR)-b-hydroxy-a-hydroxymethyl-4-nitrophenethyl] acetamide), and tetracycline (2-(amino-hydroxy-ethylidene)-4-dimethylamino-6,10,11,12a-tetrahydroxy-6-methyl-4,4a,5,5a-tetrahydrotetracene-1,3,12-trione). In total, 25 Gram-negative isolates were found to be resistant to at least one of the antibiotics tested. Some isolates were multi-drug resistant, regardless of the amount of influent the sewage treatment plant received. A Pseudomonas isolate from the smallest sewage treatment plant (approximately 2 million gallons treated per day) showed resistance to all three antibiotics, albeit at low levels (10 microg mL(- 1)). The largest number of bacteria (6 species) were isolated from the largest sewage treatment plant (45 million gallons per day). Regardless, the results of this study are in agreement with similar studies, antibiotic resistance can persist long after the antibiotics have been forgotten.
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