The efficient cyclocondensation reaction of fatty acid hydrazides with thioglycolic acid (TGA)/mercapto acetic acid and Thio lactic acid under anhydrous condition in the presence of catalyst anhydrous ZnCl2 in dimethyl formamide solvent is described. The synthesis provides the product, substituted thiazolidinone, in appropriate yield. The synthesized compounds were characterized by IR, 1 H NMR, 13C NMR, and mass spectra and these compounds were tested for in vitro antibacterial activity against the representative group of Gram-positive and Gram-negative bacteria such as Pseudomonas aeruginosa (ATCC-27853), Escherichia coli (ATCC-25922), Streptococcus pyogenes, and Klebsiella pneumonia (Clinical isolate), Methicillin resistant Staphylococcus aureus (MRSA ?ve), and also tested for antifungal activity against Aspergillus fumigates, Trichophyton mentagrophytes, Penicillium marneffei, Candida albicans. Minimum inhibitory concentration, minimum bacterial concentration, and minimum fungicidal concentration were determined for the test compounds along with the reference standards used.