Use of Molecular Markers to Compare Escherichia coli Transport with Traditional Groundwater Tracers in Epikarst.

Bacterial contamination of karst aquifers is a global concern as water quality deteriorates in the face of decreasing water security. Traditional abiotic groundwater tracers, which do not exhibit surface properties similar to bacteria, may not be good proxies for risk assessment of bacterial transport in karst environments. This study examined the transport and attenuation of two isolates of in relation to traditional groundwater tracers (rhodamine WT dye and 1-µm-diam. latex microspheres) through ∼30 m of epikarst in western Kentucky. Differential movement of the four tracers was observed, with tracer behavior dependent on flow conditions. Dye arrived at the sampling site prior to particulates. Molecular biology techniques successfully detected bacteria in the cave and showed attenuation was greater for a bacterial isolate with high attachment efficiency compared with an isolate known to have low attachment efficiency. Microspheres were first detected simultaneously with the low-attachment isolate but attained maximum concentrations during increases in discharge >11 d post-injection. Bacteria were remobilized by storm events >60 d after injection, illustrating the storage capacity of epikarst with regard to potential contaminants. The two bacterial strains were not transported at the same rate within the epikarst, showing breakthroughs during differing storm events and illustrating the importance of cell surface chemistry in the prediction of microorganism movement. Moreover, this study has shown that molecular analysis can be successfully used to target, quantify, and track introduced microbial tracers in karst terrains.

Comparing and refining karst disturbance index methods through application in an island karst setting.

The interconnected nature of surface and subsurface karst environments allows easy disturbance to their aquifers and specialized ecosystems from anthropogenic impacts. The karst disturbance index is a holistic tool used to measure disturbance to karst environments and has been applied and refined through studies in Florida and Italy, among others. Through these applications, the karst disturbance index has evolved into two commonly used methods of application; yet, the karst disturbance index is still susceptible to evaluation and modification for application in other areas around the world. The geographically isolated and highly vulnerable municipality of Arecibo, Puerto Rico's karst area provides an opportunity to test the usefulness and validity of the karst disturbance index in an island setting and to compare and further refine the application of the original and modified methods. This study found the both methods of karst disturbance index application resulted in high disturbance scores (Original Method 0.54 and Modified Method 0.69, respectively) and uncovered multiple considerations for the improvement of the karst disturbance index. An evaluation of multiple methods together in an island setting also resulted in the need for adding additional indicators, including Mogote Removal and Coastal Karst. Collectively, the results provide a holistic approach to using the karst disturbance index in an island karst setting and suggest a modified method by which scaling and weighting may compensate for the difference between the original and modified method scores and allow interested stakeholders to evaluate disturbance regardless of his or her level of expertise.

Resilience of Groundwater Impacted by Land Use and Climate Change in a Karst Aquifer, South China.

Changes of groundwater flow and quality were investigated in a subtropical karst aquifer to determine the driving mechanism. Decreases in groundwater flow are more distinct in discharge zones than those in recharge and runoff zones. Long-term measurement of the represented regional groundwater outlet reveals that groundwater discharge decrease by nearly 50% during the dry season. The hydrochemistry of groundwater in the runoff and discharge zones is of poorer quality than in the recharge zone. Indications of intensive land resource exploitation and changes in land use patterns were attributed to changes in groundwater conditions since 1990, but the influence of climate change was likely from 2001, because the water temperature exhibited increasing trends at a mean rate of 0.02 °C/yr even though groundwater depth was high in the aquifer. These conclusions imply the need for further groundwater monitoring and reevaluation to understand the resilience of aquifer during urbanization and development.

Prevalence and whole-genome sequencing characterization of Salmonella in urban karst groundwater and predominantly groundwater-fed surface waters for serotypes and antimicrobial resistance.

Karst aquifers provide a significant source of drinking water around the world; however, they are prone to bacterial contamination. We investigated the prevalence, serotypes, and antimicrobial resistance genes (ARGs) of Salmonella in urban karst groundwater and groundwater-fed surface waters. Weekly water samples (n = 443) were collected from nine groundwater sites and the Barren River in Bowling Green, KY. Additionally, cross-sectional samples were collected from 45 sites, including groundwaters and two rivers, in Tampa, FL. Salmonella was detected in 14.5% of Bowling Green samples resulting in 176 isolates representing 18 serotypes. Two (4.4%) of the Tampa samples were positive yielding eight isolates representing two serotypes. In Bowling Green, Salmonella prevalence varied by sampling site, week, month, and season, with the highest prevalence in surface water in the month of November and during summer. Salmonella Newport was the most common serotype detected. Nine of the serotypes detected in the study were among the top 20 serotypes commonly associated with human infections in the United States. Isolates were clustered with human clinical isolates, or isolates obtained from food animals, suggesting the importance of humans and agricultural practices as a source of water contamination. ARGs were detected in 12.5% (n = 176) of the Bowling Green isolates; overall, 81.8% (n = 22) of these were multidrug-resistant (resistance to ≥3 antimicrobial classes). Notably, the four Salmonella Agona isolates were resistant to four antimicrobial classes, mercury, and quaternary ammonium compounds. Seasonality and the widespread occurrence of Salmonella both in the groundwater and groundwater-fed surface waters is a potential risk to public health.

Occurrence and prevalence of antimicrobial resistance in urban karst groundwater systems based on targeted resistome analysis.

Antimicrobial resistance (AMR) is a global crisis threatening human, animal, and environmental health. The natural environment, specifically water resources, has been recognized as a reservoir and dissemination pathway for AMR; however, urban karst aquifer systems have been overlooked. This is a concern as these aquifer systems provide drinking water to about 10 % of the global population; yet, the urban influence on the resistome in these vulnerable aquifers is sparingly explored. This study used high-throughput qPCR to determine the occurrence and relative abundance of antimicrobial resistant genes (ARG) in a developing urban karst groundwater system in Bowling Green, KY. Ten sites throughout the city were sampled weekly and analyzed for 85 ARGs, as well as seven microbial source tracking (MST) genes for human and animal sources, providing a spatiotemporal understanding of the resistome in urban karst groundwater. To further understand ARGs in this environment, potential drivers (landuse, karst feature type, season, source of fecal pollution) were considered in relation to the resistome relative abundance. The MST markers highlighted a prominent human influence to the resistome in this karst setting. The concentration of targeted genes varied between the sample weeks, but all targeted ARGs were prevalent throughout the aquifer regardless of karst feature type or season, with high concentrations captured for sulfonamide (sul1), quaternary ammonium compound (qacE), and aminoglycoside (strB) antimicrobial classes. Higher prevalence and relative abundance were detected during the summer and fall seasons, as well as at the spring features. Linear discriminant analysis suggested that karst feature type had higher influence on ARGs in the aquifer compared to season and the source of fecal pollution had the least influence. These findings can contribute to the development of effective management and mitigation strategies for AMR.

Application of a cave inventory system to stimulate development of management strategies: the case of west-central Florida, USA.

The active management of air-filled cave systems is virtually non-existent within the karst landscape of west-central Florida. As in every karst landscape, caves are important because they contain a wide variety of resources (e.g., biota, speleothems) and can act as direct connections between surface and subsurface hydrological processes, potentially exacerbating the pollution of groundwater. Before sound management policies can be drafted, implemented, and enforced, stakeholders must first have knowledge of the management requirements of each cave. However, there is an informational disconnect between researchers, stakeholders, and the recreational caving community. Here, we present a cave inventory system that simplifies the dissemination of resource knowledge to stakeholders so that cave management and protection policies can be drafted and implemented at the state and local level. We inventoried 36 caves in west-central Florida, located on both public and private land, and analyzed cave resource data to provide insights on cave sensitivity and disturbance using two standardized indices. The data revealed that both public and private caves exhibit a wide range of sensitivity and disturbance, and before management strategies can be drafted, the ownership of each cave must be considered. Our inventory geodatabase serves as a link between researchers, landowners, and the public. To ensure the conservation and protection of caves, support from county or state government, combined with cave inventory data, is crucial in developing sound management policy.

Physicochemical parameters and phytoplankton as indicators of the aquatic environment in karstic springs of South China.

As the concentrated discharge outlet of an aquifer or groundwater system, a karst spring is partly independent from the aquifer, due to its formation of a pool or lake after outcropping onto the surface. Understanding how to evaluate the unique and sensitive environment of the karst spring is essential for water resource protection. Five karst springs in South China were investigated by analyzing their hydrodynamic conditions, variations in physicochemical parameters, and phytoplankton community structures. Dominated by regional or local groundwater flow, these springs had different catchment area characteristics and hydrogeological conditions. The results showed that, although they had similar water quality, their physicochemical parameters needed to be distinguished and evaluated in different ways in order to determine the cause of the observed degradation in spring water quality. Ca2+, HCO3-, and specific electrical conductivity were the major parameters reflecting the impact of regional flow from the aquifer; pH, dissolved oxygen, and water temperature indicated the local environment in and around the springs; while nitrogen and CODMn both related to the aquifer and local environment, depending on seasonal variation and human activities. The comparison of long-term nitrate data revealed that environmental pressure has increased over time. Water deterioration of Lingshui Spring was attributed to the strong interaction of surface water and groundwater. High nutrient concentrations did not correspond with the highest phytoplankton abundance or the most species. The phytoplankton community structures in the karst springs varied from place to place, depending on the hydrogeological conditions and the surrounding environment. The water environment status, as reflected by the combination of water quality indices and biological indicators, could more comprehensively represent overall water health.