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A series of 4-thiazolidinone derivatives containing 1,4-benzodioxane ring system was synthesized starting from 2,3-dihydro-1,4-benzodioxane-2-carbohydrazide. The synthesized compounds were characterized and evaluated for antibacterial activity against Staphylococcus aureus, Escherichia coli and Bacillus subtilis and antifungal activity against Aspergillus niger, Aspergillus flavus and Candida albicans by two fold serial dilution technique. Some of the synthesized compounds displayed comparable or even better antibacterial and antifungal activities than reference drugs norfloxacin, chloramphenicol and fluconazole, against tested strains.