New complexes [FeO(DMBG)]2 and [Fe(DMBG)2]Cl·0.5H2O (HDMBG: N,N-dimethylbiguanide) have been synthesized and characterized by microanalytical, IR and UV–Vis data. Electronic spectra of the complexes are characteristic for a tetrahedral stereochemistry, whilst the modifications in the IR spectra indicate the presence of the DMBG anion as chelate. The thermal decomposition investigated in air by thermal analysis confirmed the proposed formulas for the complexes and provided information concerning the modifications during heating and also the thermodynamic effects accompanying them. The thermal transformations are complex, according to TG and DTG curves including dehydration, hydrochloric acid elimination, oxidative degradation and condensation of –C=N– unit. The final product of decomposition was iron (III) oxide as powder XRD indicates. Complexes were screened for their antimicrobial properties against Gram-positive, Gram-negative bacterial, as well as fungal pathogenic strains. The results indicate that Fe(III) complexes exhibit an improved antibacterial activity against S. aureus and E. coli strains in comparison with free biguanide. The Fe(III) complexes also inhibit the ability of S. aureus, B. subtilis, P. aeruginosa and E. coli strains to colonize the inert substratum, accounting for their possible use as anti-biofilm agents.