Abstract:[Objective] We detected and analyzed β-lactam resistance in E. coli in broiler chickens in Shandong province. [Methods] The viscera samples were obtained from broiler chickens in the slaughterhouse. We screened β-lactam resistant E. coli, used the disk diffusion assay to determine antimicrobial-resistance, extracted bacterial DNA, conducted phylogenetic and biochemical assays, identified β-lactam-resistance genes and the structure of integrons and performed conjugation assays. [Results] More than 80% isolates were resistant to 3 or more β-lactams. The blaTEM and blaCTX-M resistance genes encoding class A β-lactamases have a higher rate, 86.7% and 81%, respectively, but only blaCTX-M and β-lactam resistance showed a significant correlation. The rate of β-lactam resistance genes in B1 and D2 subtypes of E. coli was higher, and the resistance was significantly enhanced, while the A0 and A1 subtypes were more sensitive. Although integrons were commonly detected in E. coli, their correlation with β-lactamase genes is low. [Conclusion] The results show that broiler-derived E. coli has highly resistant to β-lactams, and multi-drug resistance is widespread. The relationship between the resistance of β-lactams and the phylogeny in E. coli is clarified, and it provides a reference for the epidemiological surveillance of β-lactam resistance in E. coli from broilers.