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.