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The health and safety of grazing animals was the subject of microbiological monitoring on natural source of drinking waters in the upper Molise region, Italy. Surface water samples, on spring-summer season, were collected and submitted to analyses using sterile membrane filtration, cultural medium, and incubation. The level of environmental microbial contamination (Total viable microbial count, yeasts and fungi) and faecal presence (Total and faecal coliforms, <i>E. coli</i>, and <i>Salmonellae</i> spp.) were carried out. By the selective microbiological screening, twenty-three <i>E. coli</i> strains from drinking waters were isolated and submitted to further studies to evaluate antibiotic resistance by antibiograms vs. three animal and two diffuse human antibiotics. Furthermore, after a fine chemical characterization by GC and GC-MS, three Essential Oils (EOs) of aromatic plants (<i>Timus vulgaris</i>, <i>Melaleuca alternifolia</i>, <i>Cinnamomun verum</i>) aromatograms were performed and results statistically compared. The effects of EOs vs. antibiotics on <i>E. coli</i> strains isolated from drinking waters showed a total absence of microbial resistance. In our experimental conditions, even if some suggestions will be further adopted for better managements of grazing animals, because the health and safety represent a guarantee for both animals and humans.