Industrial activities release waste into aquatic sources, which is hazardous because of the pathogenic and cytotoxic behaviour of the contaminants present in the effluent. To address this issue, the citrate sol-gel auto-combustion method synthesized nickel-substituted barium M-type hexaferrites BaNixFe12−xO19 (x = 0.0, 0.1, 0.2) nanoparticles. The nanoparticles show the single-phase hexaferrite structure having a crystallite size of 52.47 to 49.30 nm with the space group P63/mmc. Magnetic study of nanoparticles indicates that increased nickel ions give rise to saturation magnetization, whereas coercivity decreases. The band gap values of the pure and Ni-doped barium hexaferrites determined by Tauc plots have been noticed to range from 1.42 eV to 1.26 eV. The photocatalytic activity of nanoparticles for methyl orange (MO) dye has been investigated. The nanoparticles have shown a dye degradation efficiency of 84.5 % within 100 min. The antimicrobial properties of the synthesized nanoparticles have also been studied. The effectiveness of synthesized nanoparticles against two types of bacteria, gram-positive Staphylococcus aureus and gram-negative Escherichia coli, as well as against two pathogenic fungi, Candida albicans and Aspergillus niger, has been investigated. The antibacterial effect has been demonstrated by all samples at all concentrations, with MIC values ranging from 1 to 4 mg/ml. Therefore, this study investigates the potential applications of nickel-doped barium M-type hexaferrites nanoparticles in real-world biological settings, suggesting their promising role in fighting against bacterial and fungal infections.