<p>Antibiotic resistance in bacteria, including the pathogenic and innocuous commensals, has become a significant global issue. The one health paradigm has highlighted the impact of antibiotic resistance among environmental bacteria and the dangers associated with their transmission to humans. Resistance genes can be transmitted vertically or horizontally across bacterial populations. The recipient bacterial populations may serve as a reservoir for these resistance genes. The occurrence of tetracycline, sulfonamide, beta-lactam, and chloramphenicol antibiotic resistance genes (ARGs) was studied in coastal Karnataka using a culture-independent method. The distribution of ARGs in water from various water bodies, including lakes, rivers, ponds, the sea, aquaculture farms, sewage, and wells, was investigated. The procedure for environmental DNA extraction to study the presence of antibiotic resistance genes was standardised, and PCR was performed for all the samples. Among the various methods used for extracting environmental DNA, the Phenol–Chloroform (PCI) method, employing different precipitating agents, yielded high-quality DNA suitable for PCR and other applications. <i>blaTEM (</i>32%), followed by <i>tetG (</i>28%) and <i>sul1 (</i>26%) were the most prevalent genes in the water samples. <i>tetB</i> was not detected in any of the samples tested. This preliminary study explored the distribution of ARGs in the aquatic environment of coastal Karnataka. The findings suggest that natural water sources, regardless of any contaminants, serve as reservoirs of antibiotic resistance determinants, and their dissemination to pathogens and opportunistic pathogens in the aquatic environment poses a threat to public health.</p>