ANAMMOX BACTERIA DETERMINATION IN THE PHARMACEUTICAL PRODUCTION WASTEWATER

Abstract

Anammox bacteria are found in sewage treatment systems, as well as in the other ecological niches, where there are anaerobic conditions. The aim of the study was to establish the ANAMMOX bacteria (Anaerobic AMMonium OXidation) presence and their systematic affiliation in active sludge samples. Methods. The ammonium, nitrite and nitrate concentration determination in the samples was obtained using spectrophotometric reactions. The ANAMMOX process presence was determined by the active sludge incubation method with mineral nutrient medium under anaerobic conditions. The anammox bacteria presence and genus affiliation were determined by FISH (fluorescence hybridization in situ) technique using specific tagged primers: the universal Tamra-Amx-0368, and also FamAmx-0820 and Fam-Kst-1 275. Results. For the anammox bacteria synthetic nutrient medium preparation there were determined the ammonium and nitrite ions concentration in the experimental test. The nutrient mineral medium was analyzed for the residual ammonium and nitrite ions concentration after cultivation. For further visual examination of the ANAMMOX bacteria, FISH reaction and the samples microscopy there were performed. Three tagged primers were used in the reaction: the universal Tamra-Amx-0368, and also Fam-Amx-0820 and FamKst-1 275. Conclusions.The decrease of the concentration of ammonium ions by 0,0395 g / l and ions nitrites at 0.0179 g / l in a synthetic nutrient medium in anaerobic conditions with gas release indicates the presence of microorganisms responsible for the ANAMMOX process. The hybridization results indicated Can. Brocadia and Can. Kuenenia presence in the sludge and water sample, in the range of 8 to 10 microcolonial units per 50 μl, as well as Kuenenia stuttgartiensis, but in smaller number, 3 to 4, of microcolonial units.