NONTUBERCULOUS MYCOBACTERIA DIVERSITY IN KARST WATERS AND BIOFILMS IN BULGARIAN CAVES

Abstract

Background: Nontuberculous Mycobacteria (NTM) are emerging pathogens causing opportunistic infections in humans and animals. Their distribution in the waters and caves of Bulgaria is poorly studied. Climatic changes are associated with changes in the amplitudes of ambient and water temperature, as well as changes in the amount of precipitation which play an essential role in the creation of reservoirs of some types of NTM in the environment. Material and Methods: We optimized the methods for successful isolation of environmental NTM and then used molecular genetic methods for identification. Results: A total of 235 samples (karst water, sediments, soil, bat guano) were collected in some caves of the following karst regions: 203 in Vratsa Karst area, 204 in Ponor Karst area, 205 in Bezdenski area and 303 in Karst and caves of Bosnek region. Primary isolation of mycobacteria by Löwenstein–Jensen at room temperature was more successful than on liquid media at 37°C. We identified NTM in 10% (n=24) from these materials. Diverse NTM included: M. chelonae (n=3), M. gordonae (n=2), M. intermedium (n=3), M. scrofulaceum (n=1), M. szulgai (n=4), M. fortuitum group (n=4), NTM mix culture (n=5), M. terrae complex (n=1), Mycobacterium sp. (n=1). Rapidly growing NTM (M. chelonae, M. fortuitum group) were the most common. The isolates belonged to group of environmental saprophytes (Risk group 1) and potential pathogens (Risk group 2). Conclusions: We successfully implemented a procedure for decontamination and isolation of NTM from the environment. For the first time in the country, NTM species were identified in biofilms, karst waters, soil and bat guano within caves. The presence of NTM in cave ecosystems represents a potential source for human infection.