Detection of mycobacteria in the environment of the Moravian Karst (Bull Rock Cave and the relevant water catchment area): the impact of water sediment, earthworm castings and bat guano

Abstract

ABSTRACT:The presence of mycobacteria was studied in Bull Rock Cave ("Byci skala") and the water catchment area of Jedovnice Brook ("Jedovnicky potok") using direct microscopy after Ziehl-Neelsen (ZN) staining, culture examination and molecular techniques. Mycobacteria were detected in 47.1% of a total of 68 samples. The mycobacterial genes hsp65 and dnaA were detected and sequenced in 37 (74.0%) out of the 50 cave environmental samples and in 10 (55.6%) out of the 18 samples of water catchment sediments. Nine species of slowly growing mycobacteria (M. terrae, M. arupense, M. gordonae, M. lentiflavum, M. parascrofulaceum, M. parmense, M. saskatchewanense, M. simiae and M. xenopi) and two subsp. (M. avium subsp. avium and M. avium subsp. hominissuis) were detected. Fourteen species of rapidly growing mycobacteria (M. chelonae, M. chubuense, M. poriferae, M. flavescens, M. fortuitum, M. porcinum, M. rhodesiae, M. gilvum, M. goodii, M. peregrinum, M. mageritense, M. vanbaalenii, M. gadium and M. insubricum) were detected. The highest mycobacterial presence was documented by ZN staining and/or culture examinations in earthworm castings and bat guano (73.3% positivity out of the 15 samples) in the cave environment and in the water sediments collected under the outflow from the wastewater treatment plants (77.8% positivity out of nine samples). The highest total organic carbon (TOC) was detected in wooden material and earthworm castings with pH values between 5.0 and 7.7 in the cave environment and in water sediments collected under the outflow from the wastewater treatment plants with pH between 5.8 and 7.0. It could be concluded that the karst cave environment with its running surface water contaminated with different microorganisms or chemical substances creates favourable conditions not only for animals (especially earthworms) but also for mycobacteria. This fact is also demonstrated by the presence of these mycobacteria in the cave environment mainly in earthworm castings and bat guano.

Influence of Human Activity on Surface Water Quality in Moravian Karst

“…1). The Jedovnice Brook flows through the cave system of Rudice Swallow outside the basin and springs to the surface again behind Bull Rock Cave after 5.8 km [6]. The PLA Moravian Karst was established in 1956.…”

“…In karst areas and areas communicating through the aquatic environment with these areas, it is necessary to pay special attention not only to the danger of discharging all waste treated and untreated water from settlements and recreational sites but also to the proper management of agricultural land. It is recommended to exclude the amelioration accelerating runoff, dumping waste in inappropriate places and the application of chemicals, including pesticides [6,28].…”

“…Significant contamination with organic substances and nutrients was observed. The contamination leads to water eutrophication and represents numerous hazards both on the surface and in the karst underground [5,6].…”

Development of Population and Technical Infrastructure in Municipalities of the Moravian Karst: Potential Influence on the Organic Pollution of Water

“…Vyvstává proto otázka, jaké jsou hlavní zdroje organického znečištění vody (povrchové a krasové) v současné době a jak k němu přispívá činnost lidí a rozsah urbanizovaného území. Z předešlých ekologických studií prováděných v rámci sledování environmentálních a podmíněně patogenních mykobakterií v MK ve vodním prostředí vyplývá, že je významné organické znečištění nejenom přímo v prostoru CHKO MK, ale také v celém povodí vodních toků ústících do MK (Modra et al, 2016, Pavlík a kol., 2016a. Komunální odpadní voda se v současné době jeví jako jeden z nejvýznamnějších rizikových faktorů při eutrofizaci povrchových vod a při šíření různých původců infekčních i neinfekčních onemocnění lidí a zvířat v prostředí ekonomicky rozvojových i rozvinutých zemí (Pavlík, 2014(Pavlík, , 2016.…”

“…V povrchových tocích a ve vodě v krasovém prostředí jsou nyní často zjišťovány intenzivní kontaminace organickými složkami, které vedou k eutrofizaci vody a představují jak na povrchu, tak v krasovém podzemí značná zdravotní pobytová rizika (Pavlík a kol., 2016a, Modra et al, 2016.…”