The aim of this study was to identify bacteria present in ripened manganese removal filters for drinking water production. The bacterial population was identified with ‘next generation’ DNA sequencing, and specific bacteria were quantified with quantitative polymerase chain reaction (qPCR) and characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis. The ‘next generation’ DNA sequencing analysis showed a bacteria population shift from the iron oxidizing species Gallionella spp. in the Fe-filter to manganese and nitrite oxidizing species Pseudomonas spp. and Nitrospira spp., respectively, present in the manganese removal filter. qPCR analysis confirmed the presence of a low concentration of the well-known Mn2+-oxidizing species Ps. putida in the manganese removal filter backwash water. Bacteria of the genus Pseudomonas, isolated from backwash water from a manganese removal filter were cultured and identified with MALDI-TOF MS analysis. Amongst others, P. gessardii, P. grimontii, and P. koreensis were identified. The presence of several manganese oxidizing bacteria species in ripened filter media supports the assumption that a microbial consortium is involved in the oxidation of manganese. Understanding the mechanisms by which manganese coating of filter media commences could endorse the creation of conditions favouring Birnessite formation, and possibly help in reducing typically long ripening periods of manganese removal filters with virgin filter media.

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