Chlorhexidine bigluconate (CНX) is widely used as a disinfectant, but it is not effective against spore-forming microorganisms, as well as viruses. In this work, a method has been found to increase the biocidal activity of chlorhexidine by using it as part of a complex including ethylenediaminetetraacetic acid (EDTA) and zinc chloride. The structure of the three-component complex СНX-EDTA-zinc is proved by the MALDI-MS method. The biocidal activity of the chlorhexidine complex has been studied in vitro and in vivo experiments. It is shown that the complex is significantly superior to chlorhexidine alone, both in terms of activity level and in the breadth of biocidal action. In relation to the studied bacterial and fungal strains, the СНX-EDTA-Zn complex was 4–5 times more active than chlorhexidine bigluconate. In concentrations from 1.0 mg/ml to 0.008 mg/ml (depending on the type of micro-organism), in vitro the complex showed both bacteriostatic and bactericidal effects against the main pathogens of bacterial diseases of birds. In clinical conditions, the complex has shown high efficiency in the treatment of dermatitis in small domestic and farm animals. Also, in vitro and in vivo, the complex showed unexpectedly high antitubercular activity comparable to that of monofloxacin, including on drug-resistant strains of mycobacteria. in vitro experiments involving polio virus and adenovirus have shown that the СHX-EDTA-Zn complex possesses virulent action.