Abstract:720 samples of feces, soil and sewage were collected from 12 broiler farms in Huzhou, Jiaxing and Shaoxing cities in this study. 234 isolates of Escherichia coli were identified by 16s rRNA sequencing, antibiotic-resistant phenotypes were determined by K-B (Kirby-Bauer), antibiotic-resistant genotypes and integrons were identified by PCR. The transfer risk of antibiotic resistance of Escherichia coli was analyzed by co-occurrence network. 234 isolates of Escherichia coli were highly resistant to cefazolin, azithromycin, tetracycline and sulfonamides, and the resistance rate of isolates in Jiaxing to 17 antibiotics was higher than that in the other two cities. The results of genotype identification showed that the detection rate of aadA1 was the highest among the 234 strains of Escherichia coli, which was 88.89%, followed by tet(A), sul1 and blaTEM, respectively. Among the 29 antibiotic resistance genes, the detection rate of 18 genes in Jiaxing was higher than that in the other two cities. The detection rates of type I integrase in Huzhou, Jiaxing and Shaoxing were 28.38%, 40.96% and 32.47%, respectively. The results showed that the antibiotic resistance rate and the detection rate of antibiotic resistance genes of isolates in Jiaxing were relatively high. Co-occurrence network analysis showed that there was a high risk of co-transfer of qnrB and mph(A), fosA and blaTEM, fosA and sul1, sul2 and aadA1 in E. coli in Jiaxing. In summary, this study investigated antibiotic resistance and transmission risk in three cities of Zhejiang Province, providing scientific guidance for ensuring animal food safety and monitoring key specific antibiotic resistance genes.