In this study, the effect of Carica papaya seed macerate on conjugal R plasmid transfer from Salmonella typhimurium to Escherichia coli was investigated in vitro and in the digestive tract of gnotobiotic mice. Twenty-five micrograms per milliliter and 430 mg (administered intragastrically twice a day) of papaya seed macerate concentrations were used during conjugation for in vitro and in vivo assays, respectively. High frequency of conjugation inhibition by macerate was observed for both in vitro and in vivo experiments, independently of bacterial growth and mating conditions. Papaya seed macerate caused a reduction of the transconjugant population ranging from 71% to about 100%. There was no lethal effect of the seed macerate on donor or recipient cells in the concentrations used. Once the mechanisms and magnitude of resistance gene transfer are clearly understood, strategies to reduce or minimize the dissemination of these genes could be relevant. The data here obtained show a clinical potential use of papaya seed macerate on this transfer.