Toxicological Study of Fluorescent Hydrologic Tracer Dye Effects on Cave Bacteria.

Fluorescent dyes are commonly used as hydrologic tracers in a variety of surface and subsurface environments, including karst aquifers and caves, but the fragile nature of karstic groundwater ecosystems suggests a cautious approach to selecting dyes. This study tested the effects of four fluorescent dye tracers (uranine, eosin, pyranine, sulforhodamine B) on microorganisms from Fort Stanton Cave, New Mexico, United States. Toxicity of the dyes was tested on bacteria isolated from the cave and on a sediment sample collected adjacent to Snowy River in Fort Stanton Cave. The isolates showed minimal inhibition by the four dyes in an agar diffusions assay. Minimum inhibitory concentrations calculated from liquid culture assays of one isolate were 35 g/L for uranine, 3.5 g/L for eosin, 0.1 g/L for pyranine, and 10 mg/L for sulforhodamine B. A 14 C-glucose radiotracer experiment showed zero inhibition of overall microbial activity in a sediment sample at all dye concentrations, except at 350 g/L eosin. Thus, there are no cave-specific findings to indicate that Fort Stanton's microbes are especially sensitive to these commonly used dyes. Moreover, a literature survey of mutagenicity tests on these dyes indicates they are safe for environmental use. These results corroborate previous dye toxicity tests and suggest that these four dyes are suitable for use at Fort Stanton Cave in the concentration ranges commonly used for groundwater tracing. While broader testing of dyes with microbes from other caves is advised, the results suggest the dyes may be safe for all karst aquifers.

Lighting Effects on the Development and Diversity of Photosynthetic Biofilm Communities in Carlsbad Cavern, New Mexico.

Photosynthetic cave communities ("lampenflora") proliferate in Carlsbad Cavern and other show caves worldwide due to artificial lighting. These biofilms mar the esthetics and can degrade underlying cave surfaces. The National Park Service recently modernized the lighting in Carlsbad Cavern to a light-emitting diode (LED) system that allows adjustment of the color temperature and intensity. We hypothesized that lowering the color temperature would reduce photopigment development. We therefore assessed lampenflora responses to changes in lighting by monitoring photosynthetic communities over the course of a year. We measured photopigments using reflected-light spectrophotometric observations and analyzed microbial community composition with 16S and 18S rRNA gene amplicon sequencing. Reflected-light spectrophotometry revealed that photosynthetic biofilm development is affected by lighting intensity, color temperature, substrate type, and cleaning of the substrate. Gene sequencing showed that the most abundant phototrophs were Cyanobacteria and members of the algal phyla Chlorophyta and Ochrophyta At the end of the study, visible growth of lampenflora was seen at all sites. At sites that had no established biofilm at the start of the study period, Cyanobacteria became abundant and outpaced an increase in eukaryotic algae. Microbial diversity also increased over time at these sites, suggesting a possible pattern of early colonization and succession. Bacterial community structure showed significant effects of all variables: color temperature, light intensity, substrate type, site, and previous cleaning of the substrate. These findings provide fundamental information that can inform management practices; they suggest that altering lighting conditions alone may be insufficient to prevent lampenflora growth.IMPORTANCE Artificial lighting in caves visited by tourists ("show caves") can stimulate photosynthetic algae and cyanobacteria, called "lampenflora," which are unsightly and damage speleothems and other cave surfaces. The most common mitigation strategy employs bleach, but altering intensities and wavelengths of light might be effective and less harsh. Carlsbad Cavern in New Mexico, a U.S. National Park and UNESCO World Heritage Site, has visible lampenflora despite adjustment of LED lamps to decrease the energetic blue light. This study characterized the lampenflora communities and tested the effects of color temperature, light intensity, rock or sediment texture, and time on lampenflora development. DNA amplicon sequence data show a variety of algae and cyanobacteria and also heterotrophic bacteria. This study reveals microbial dynamics during colonization of artificially lit surfaces and indicates that while lowering the color temperature may have an effect, management of lampenflora will likely require additional chemical or UV treatment.