Three hundred and thirty-seven isolates of Salmonella Pullorum from eastern China between 1962 and 2010 were characterized for antimicrobial susceptibility (disk diffusion method), the presence of integrons (polymerase chain reaction followed by sequencing) and the ability to form biofilms (semi-quantitative adherence assay). Two hundred and fifty-eight isolates (76.6%) exhibited multiple drug resistance (MDR; resistant to at least three different classes of antimicrobials), and the level of drug resistance is increasing with time. There were three isolates (9.4%) exhibiting MDR from 1962 to 1968. MDR rates began to increase for isolates between 1970 to 1979 and 1980 to 1987 (64.6 to 78.7%). The MDR rates reached 96.6% for isolates between 1990 and 2010. Polymerase chain reaction screening for integrons showed that 75 isolates (22.3%) were positive for class 1 integrons while none were positive for class 2 integrons. All of the class 1 integron-positive isolates exhibited MDR and were more frequently resistant than the negative isolates. Two hundred and twenty isolates (65.3%) had the ability to form biofilms, and bacterial resistance levels to cefamandole, trimethoprim and trimethoprim/sulfamethoxazole were significantly higher for biofilm-positive groups than the biofilm-negative groups. Our data show that multidrug resistance is common among S. Pullorum isolated from eastern China, being more frequent after 1990 than before 1990, and the presence of class 1 integrons is associated with multidrug resistance.