Aim. The aim of the study was to consider the possibility of using the consortia of chemolithoautotrophic ferrobacteria from Gallionella genus and heterotrophic bacteria from Lepthothrix genus for the biological method of groundwater purification. Methods. The photocolorimetric method to determine the concentrations of ammonium and iron ions, the titrimetric method to determine the hydrocarbon and total alkalinity, the method of determining the permanganate oxidizability using the Kubel method, the potentiometric method to determine the values of рН and Еh, the electronic microscopy using the X-ray spectral analysis of matrix structures of bio-minerals, microbiological and statistical methods. Results. The main technological parameters of the water deironing process were defined as follows: the filtration velocity of the bioreactor – 7–11 m/h, and of the filters – 3.5–5 m/h; the filter-cycle duration – 48 h. It was found that the application of the two-stage technology of biological deironing in the bioreactor and filters provided for the possible removal of Fe2+ compounds up to 5.0 mg/cdm, ammonium nitrogen — up to 1.5 mg/cdm, soluble organic substances by PO – up to 6.0 mg O2/cdm. It was determined that the optimal parameters for the process of biological purification of neutral groundwaters, containing increased concentrations of Fe2+ cations were as follows: рН 7.0–7.2; hydrocarbon alkalinity 2.5–2.2 mmol/cdm; content of soluble oxygen – 1.5–2.0 mg/cdm. The ability of concentrated (Dos 200 mg/cdm) matrix structures of Gallionella and Lepthothrix ferrobacteria to remove Cr6+ ions from natural groundwaters was determined. The study found no considerable differences in the efficiency of applying disinfectants, produced using polyhexamethylene guanidine chloride (PHMGchl) or polyhexamethylene biguanidine chloride (PHMBchl). In concentrations of 0.25–0.5 %, they effectively disinfect pathogenic microorganisms, including Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. The same was found true for the bacteria, most common in the systems of technical reverse water supply, the systems of water circulation, plant watering, and fire tanks, including Aeromonas hydrophila, Aeromonas salmonicida, Pseudomonas sp., E. coli, Flavobacterium columnare, Micrococcus lysodeikticus. Conclusions. This was the first study on the possibility of applying the biotechnology of groundwater purification from excessive amounts of iron in conditions of uneven hydraulic burden, notable for the water supply systems of rural areas and most agricultural enterprises in the north-western and northern regions of Ukraine. The specificities and perspectives of PHMGchl application in the systems of water preparation were studied. It was found that from the standpoint of safety and efficiency, the use of the water deironing processes involving iron bacteria was reasonable in the systems of water consumption and technical water supply, and the disinfection of water using PHMG was possible only in the second case, due to some toxicity of the preparation. The optimal parameters for the process of biological purification of neutral groundwaters, containing increased concentrations of Fe2+ cations were determined.