The freshwater macroalgae (Cladophora glomerata), a filamentous green alga, collected from the sub-branch of the dam canal, was identified based on morpho-anatomical characters and chemical compositions. The algal biomass containing carbohydrate, protein, lipid, contents were 31.65± 1.5, 13.7± 1.5 and 2.05± 1.5 wt.%, respectively. Also, C. glomerata has chlorophylls and carotenoids pigments were 258.35±2.16 and 79.54±0.67 μg/ml, respectively. Furthermore, to evaluate the antimicrobial potential of methanolic solvent extracts derived from C. glomerata against bacteria and fungus. The feasibility of utilizing C. glomerata extracts as a potential source for the antibacterial, antifungal properties was investigated. The methanol solvent extracts of C. glomerata used in this article have revealed significant bacterial strains, namely, B. substilis, K. pneumonia, P. aeruginosa, P. vlgaris, S. aureus and E. coli. The Epidermophyton floccosum shows that higher inhibition was reached to 91.02 %. Fungal strains C. albicans, E. floccosum, M. canis, M. phaseolina, P. oedochilum, R. solani, A. flevus, N. oryzae and T. hamatum were examined with C. glomerata extracts. Staphylococcus aureus express that highed positive control Ampicillin compared to other fungus species in this article. These results suggest that the methanol extracts of C. glomerata possess an effective wide spectrum of antimicrobial and antifungal, which can assist as a remarkable source for antimicrobial compounds.