Comparative microbial sampling from eutrophic caves in Slovenia and Slovakia using RIDA ®COUNT test kits

Abstract

RIDA®COUNT test plates were used as an easy-to-handle and rapid indicator of microbial counts in karst ecosystems of several caves in Slovakia and Slovenia. All of the caves had a high organic input from water streams, tourists, roosting bat colonies or terrestrial surroundings. We sampled swabs, water and air samples to test robustness and universality of the RIDA®COUNT test kit (R-Biopharm AG, Germany, http://www.r-biopharm.com/) for quantification of total bacteria, coliforms, yeast and mold. Using data from swabs (colony-forming units CFU per cm 2 ) we proposed a scale for description of biocontamination level or superficial microbial load of cave niches. Based on this scale, surfaces of Ardovská Cave, Drienovská Cave and Stará Brzotínská Cave (Slovakia) were moderately colonized by microbes, with total microbial counts (sum of total bacterial count and total yeast and molds count) in the range of 1,001-10,000 CFU/100 cm 2 , while some surfaces from the show cave Postojna Cave (Slovenia) can be considered highly colonized by microbes (total microbial counts ≥ 10,001 CFU/100 cm 2 ). Ardovská Cave also had a high concentration of airborne microbes, which can be explained by restricted air circulation and regular bat activity. The ratio of coliform to total counts of bacteria in the 9 km of underground Pivka River flow in Postojna Cave dropped approximately 4-fold from the entrance, indicating the high anthropogenic pollution in the most exposed site in the show cave. The RIDA®COUNT test kit was proven to be applicable for regular monitoring of eutrophication and human influence in eutrophic karst caves.

Aerosolized Microbes from Organic Rich Materials: Case Study of Bat Guano from Caves in Romania

“…The counts of bacteria after 48 and for fungi after 72 hours were considered for statistical analyses. Prolonged incubation for 24 hours gives a more realistic view of the microbial communities, as some cave microbes have demonstrated slow growth on RIDAHCOUNT media (Mulec et al, 2012a). Isolates that exhibited ß-D-glucuronidase and ß-Dgalactosidase biochemical activities on RIDAHCOUNT E. coli/Coliform plates were considered indicative of E. coli (R-Biopharm AG, Germany).…”

Sustained Anthropogenic Impact in Carter Saltpeter Cave, Carter County, Tennessee and the Potential Effects on Manganese Cycling

“…This hypothesis is bolstered by several field studies investigating nutrient loading in caves via tourism (Ikner et al, 2007), septic effluent (Simon and Buikema, 1997), and input of rich carbon sources such as guano, feces, and human traffic (Mulec et al, 2012) where, in all three studies, cultivable counts of bacteria (either total CFUs, total aerobic …”

ATP luminescence assay as a bioburden estimator of biomass accumulation in caves

“…Surfaces contaminated with animal excrement, e.g. from bats (Mulec et al, 2012a) and the presence of visible microbial biofilms (Mulec et al, 2015) are important sources of microbial biomass (Table 3) in the underground. Some caves do not rely only upon the input of organic matter from the surface, but are characterized by in situ microbial biomass production based on chemoautotrophic metabolism (Jones & Macalady, 2016).…”

“…UltraSnap TM kit has previously been used for surface swabbing in Lechuguilla Cave (New Mexico, USA), where the DAPI total cell count did not change significantly in line with changes in ATP levels (Johnston, 2013). In a previous study in Postojna Cave (Mulec et al, 2012a), similar surfaces were swabbed using RIDA®COUNT test plates directly for swabbing. The swabbing procedure described in this study and the use of comparable nutrientrich media (NA vs. RIDA®COUNT Total for bacteria and MEA vs. RIDA®COUNT Yeast&Mold Rapid for fungi), with similar cultivation conditions (35°C and 37°C for 48 hours for bacteria, 20°C for 72 hours for fungi) resulted in 3-times up to 80-times higher CFU retrieval.…”