We investigated the potential anti-microbial and anti-biofilm activities of previously synthesized 1,3-dioxolane derivatives against several pathogen microorganisms. In vitro antibacterial activities of new compounds against Staphylococcus aureus ATCC 29213, S. epidermidis ATCC 12228, Enterococcus faecalis 29212, Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 4352, Proteus mirabilis ATCC 14153 and antifungal activities against Candida albicans ATCC 10231 were investigated. The minimum inhibitory concentrations (MIC), and minimum biofilm eradication concentrations (MBEC) were determined by microbroth dilution technique. As a result, two molecules were found active against the planktonic and biofilms of the E. faecalis and S. epidermidis. The MIC and MBEC values of those two agents were 9.76 -39.06 mg/L and 2000 -5000 mg/L, respectively. In addition, they inhibited the biofilm attachment according to time, and they showed the significant inhibitor activity against biofilm formation at 24 h according to their concentrations. According to these results, two 1,3-dioxolane derivatives were active against the planktonic and biofilm cells of some bacteria.