Background: One of the major causes of pathogenic E. coli is the feces of infected livestock, and the management of the livestock environment is necessary to prevent pathogenic E. coli . Objectives: The prevalence of pathogenic E. coli was identified from livestock environments, and the molecular characteristics and antibiotic resistance profiles of the isolated pathogenic E. coli strains were analyzed. Methods: In 2022 and 2023, nine points of livestock houses at sites in Chungcheongnam-do Province were selected, and 100 cow feces or soil samples around the livestock houses were collected once per month. Pathogenic E. coli was isolated by selective culture and identified using multiplex PCR. Antibiotic resistance was tested on the isolated strains by using VITEK-2, and candidate strains were selected to perform 16s rRNA sequencing and phylogenetic analysis. Results: A total of 100 samples were tested, and 60 pathogenic E. coli strains were isolated. Of these, 45 and 15 isolates were determined to be single and hybrid pathogenic E. coli , respectively. Among the 15 hybrid pathogenic E. coli strains, eight, five, and two strains were respectively identified as EHEC/ETEC, EHEC/ EPEC, and EHEC/ETEC/EPEC hybrids. All 45 isolates showed resistance to at least one antibiotic, and they were susceptible to cefotaxime, amikacin, nalidixic acid, and ciprofloxacin. The highest resistance was against cefalotin, tetracyclin, and ampicillin (20.0%~58.3%). The 16s rRNA sequences of candidate isolates revealed nucleotide sequence identities of 99.1% to 100%. Conclusions: In order to manage pathogenic E. coli from the One Health animal environment perspective, the characteristics of the occurrence of pathogenic E. coli from the livestock environment and molecular biology and antibiotic resistance to isolated strains were analyzed. In order to prevent and manage the occurrence of pathogenic E. coli , these monitoring studies must be continuously conducted.