Various products are observed in biological oxidation and reduction of molecules containing elements of variable valence. The variability is caused by the diversity of microorganisms and their metabolic enzymes, which may develop into novel processes in wastewater treatment. The study aimed to develop a novel denitrification process forming nitrite and ammonium in wastewaters containing thiocyanate. High-efficiency nitrite and ammonium production was observed due to autotrophic partial denitrification and ammonification as a result of nitrate and thiocyanate removal. Nitrite, ammonium and sulfate were observed as the ultimate products. The increased NO3--N/SCN--N ratio in the treated wastewater resulted in the decreased removal efficiency of nitrate, and the increased nitrate-to-nitrite transformation ratio and the ratio of NO2--N to NH4+-N. Thiocyanate sulfur was oxidized to sulfate via intermediate elementary sulfur providing electron to nitrate or nitrite. The Thiobacillus genus dominated in the sludge providing ammonium and nitrite as substrate for the potentially anammox process.