π-Conjugated azomethine ligands differing in the naphthalene or phenylmethane-centered core structure and their divalent cobalt, nickel, copper, and zinc metal complexes were prepared and well-characterized by spectral analyses in solid state. Magnetic natures of the complexes were determined by magnetic susceptibility measurements in solid-state. Their remarkable photophysical characteristics were recorded by Uv–vis and Fluorescence spectroscopic techniques. At their excitation wavelenght of 265 nm, all molecules exhibited triple fluorescence emission bands with promising intensities above 673 nm in near infra-red region. Antibacterial and antibiofilm activities of the π-conjugated azomethines are promising for potential applications in medical and healthcare settings. Hence, the antibacterial/antibiofilm activity of the π-conjugated azomethine ligands and their metal complexes against some clinically important bacteria namely Staphylococcus aureus (MSSA), Methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa and Proteus mirabilis was investigated, and the obtained results have shown that the ligands and complexes had a remarkable antibacterial effect, especially on Proteus mirabilis. Metal complexes have been found to have a significant inhibitory effect on biofilm formation by MRSA, MSSA, and P. mirabilis compared to ligands. The copper (II) complex of ligand-2 showed the highest inhibition percentage, significantly reducing biofilm formation for MRSA and MSSA. Furthermore, cobalt (II) complexes of the ligands selectively inhibited the growth of the opportunistic pathogen P. mirabilis biofilms, indicating that metal complexes might be a good choice for future antibiofilm studies.