Pyrrolidine dithiocarbamate (PDTC), an antioxidant with a metal-chelating activity, has been used widely to inhibit the expression of inflammatory genes in vitro and in vivo. This study investigated whether PDTC has an antimicrobial activity against various bacteria. The antibacterial activity of PDTC and other compounds was evaluated in vitro by the broth microdilution method against Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Staphylococcus aureus, and Escherichia coli. Bacterial growth was inhibited by PDTC, where a wide range of sensitivity was demonstrated among the tested bacteria. The antibacterial activity of PDTC was reduced by the addition of copper chloride; in contrast, it was enhanced considerably by zinc chloride. Two different zinc chelators, Ca-saturated EDTA (Ca-EDTA) and N,N,N',N'-tetrakis (2-pyridylmethyl) ethylenediamine, blocked the antibacterial activity of PDTC, whereas Zn-EDTA failed to reduce the activity of PDTC. These results demonstrate for the first time that PDTC possesses an antibacterial activity, for which zinc is required, and suggest that PDTC, possessing a dual anti-inflammatory and antibacterial activity, may be considered for topical use for inflammatory diseases of bacterial origin.