To investigate the prevalence and characteristics of ciprofloxacin and cefotaxime co-resistant Salmonella isolates in broiler flocks.Methods Ciprofloxacin and cefotaxime co-resistant Salmonella isolates were selected from isolates recovered from cloaca or rectal swabs and environment samples collected from 4 different districts in Henan Province. Ciprofloxacin and cefotaxime co-resistant Salmonella isolates were subjected to antimicrobial susceptibility testing, phylogenetic analysis and further characterized by screening for β-lactamase genes and quinolone resistance determinants by PCR and followed by DNA sequence analysis. Results Totally, five ciprofloxacin and cefotaxime co-resistant Salmonella isolates were recovered from 52 isolates, which all belonged to Salmonella Indiana. There were two antimicrobial resistant profiles which were AMP-CAZ-CHL-CIP-CTX-GEN (n=1) and AMP-CAZ-CHL-CIP-CTX-GEN-SXT-TET (n=4). Point mutations in topoismerase encoded genes gyrA and parC were identified, and all isolates carried plasmid-mediated quinolone resistance genes, including oqxAB and aac(6′)-Ib-cr, but qnrA, qnrB, qnrS, qnrC, qnrD and qepA were not detected. All of the five cefotaxime-resistant Salmonella were due to the production of plasmid borne blaCTX-M-65.Conclusion The extensive prevalence, complicated quinolone resistance mechanisms of isolates and the possible transmission of blaCTX-M-65 were existed in broiler flocks and environment. In order to clarify the farm-table-patient resistant relevance chain of infection transmission and to provide a scientific basis for antibiotic medication in clinical, the continuous surveillance of multidrug-resistant Salmonella in broiler flocks should be carried out.