Fulvic acid (FA) is a heterogeneous mixture of organic macromolecules found in the waters, soils, and sediments of the earth's surface. The ability of electrospray ionization (ESI) to effectively transfer large ions from the solution phase to the gas phase and the coupling of ESI to the high-mass-resolution capabilities of Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) provide a potential method for the mass spectrometric analysis of FA. Positive- and negative-ion ESI FT-ICR MS analyses of four reference International Humic Substances Society FAs were performed. The spray solution composition was found to have a dramatic effect on the ion distributions, with high-mass aggregates (m/z approximately 2000-4000) being formed in less polar spray solutions. Positive-ion spectra for each FA obtained under optimum conditions resulted in number-average molecular weights ranging from 1700 to 1900. The mass spectra were extremely complex, with ion distributions on the order of m/z approximately 500-3000. The presence of more than one ion at each nominal mass was routinely observed. Negative-ion ESI analysis of the FA samples resulted in the observation of multiply charged ions whose distributions could be affected by the acidification of the spray solution. Solution parameters which have been reported to affect molecular weight distributions of FA such as pH, ionic strength, and concentration of multivalent cations were found to have little or no effect on the observed m/z distributions.
Caves are often assumed to be static environments separated from weather changes experienced on the surface. The high humidity and stability of these subterranean environments make them attractive to many different organisms including microbes such as bacteria and protists. Cave waters generally originate from the surface, may be filtered by overlying soils, can accumulate in interstitial epikarst zones underground, and emerge in caves as streams, pools and droplets on speleothems. Water movement is the primary architect of karst caves, and depending on the hydrologic connectivity between surface and subsurface, is the most likely medium for the introduction of microbes to caves. Recently published metabarcoding surveys of karst cave soils and speleothems have suggested that the vast majority of bacteria residing in these habitats do not occur on the surface, calling into question the role of microbial transport by surface waters. The purpose of this study was to use metabarcoding to monitor the aquatic prokaryotic microbiome of a cave for 1 year, conduct longitudinal analyses of the cave’s aquatic bacterioplankton, and compare it to nearby surface water. Water samples were collected from two locations inside Panel Cave in Natural Tunnel State Park in Duffield, VA and two locations outside of the cave. Of the two cave locations, one was fed by groundwater and drip water and the other by infiltrating surface water. A total of 1,854 distinct prokaryotic ASVs were detected from cave samples and 245 (13.1%) were not found in surface samples. PCo analysis demonstrated a marginal delineation between two cave sample sites and between cave and surface microbiomes suggesting the aquatic bacterioplankton in a karst cave is much more similar to surface microbes than reported from speleothems and soils. Most surprisingly, there was a cave microbe population and diversity bloom in the fall months whereas biodiversity remained relatively steady on the surface. The cave microbiome was more similar to the surface before the bloom than during and afterwards. This event demonstrates that large influxes of bacteria and particulate organic matter can enter the cave from either the surface or interstitial zones and the divergence of the cave microbiome from the surface demonstrates movement of microbes from the epikarst zones into the cave.
The geology of Great Smoky Mountains National Park (GRSM) in Tennessee and North Carolina is dominated by siliciclastics and metamorphic strata. However, in the western portion of GRSM, a series of carbonate fensters (windows) expose the Lower Ordovician–age section of the Knox Group, a series of dolomite and limestone units that are partially marbleized as a result of contact metamorphism from the Great Smoky fault. The fensters create opportunities for allogenic recharge to occur at points along the contact of the surrounding insoluble strata with the underlying soluble carbonates. The combination of chemically aggressive surface recharge and vertical relief has resulted in the formation of deep caves, many of which have active streams and water resources. Though the karst is limited in extent and the number of caves is fairly small, the significance of the resources is substantial, with several of the caves in the area over 150 m in depth and at least two being major bat hibernacula. In 2017, the U.S. Geological Survey (USGS) began a study to better understand the hydrologic behavior of these karst systems through hydrologic and geochemical monitoring, groundwater tracing using fluorescent dyes, and seepage runs. Stage and water-quality instrumentation was installed in two caves in GRSM, the main stream of Bull Cave, and in a sump pool in Whiteoak Blowhole, at 173 m and 70 m below land surface, respectively. Following setup of the cave sites, dye injections were conducted to determine discharge points for four of the deep cave systems on Rich Mountain and Turkeypen ridge. Results show water in these systems has an extremely rapid travel time, with tracers detected from caves to springs in less than 24 h for each of the systems. This field guide describes the complex geology, regional hydrogeology, and unique landscape characterized by high-gradient subterranean streams, carbonate fensters, and deep caves of the GRSM karst.
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