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The dynamics of organic carbon (OC), turbidity, faecal indicator bacteria and physicochemical parameters was studied in a karst system near Yverdon, Switzerland. Online measurements and sampling were done at a swallow hole draining an agricultural surface (the input), and two groups of springs (the outputs) that often show bacterial contamination. A fluorescent tracer that was injected into the swallow hole during low-flow conditions first arrived at the springs 10-12 days after injection; the total recovery rate was 29%. Previous tracer tests during high-flow conditions gave shorter travel times. After a major rainfall event, a primary turbidity peak was observed at the springs. It coincides with the rising limb of the hydrograph, indicating remobilisation of autochthonous particles from the aquifer. A secondary turbidity peak occurs several days later, suggesting the arrival of allochthonous particles from the swallow hole. Wider peaks of OC and bacteria were observed simultaneously. Applying methods from molecular microbiology (PCR-DGGE) allowed characterisation of the bacterial communities at the swallow hole and the springs. The results demonstrate that the swallow hole is an important source of groundwater contamination, while its contribution to aquifer recharge is insignificant. OC appears to be a better indicator for bacterial contamination than turbidity. En période d'étiage, un essai de traçageà l'uranine aété réalisé depuis la perte. Le traceur est apparu aux sources 10-12 jours après l'injection; la masse de restitution totale aété de 29%. Des essais précédents, réalisés en hautes eaux, ont montré des temps de transit plus court. Suitè a unévénement pluvieux important, un pic de turbidité primaire, synchrone avec l'augmentation du débit, est observé aux sources, indiquant une re-mobilisation des sédiments autochtones de l'aquifère. Un pic de turbidité secondaire apparaît quelques jours plus tard aux sources, suggérant l'arrivée de matériel allochtone de la perte. Cette dernière est accompagnée de pics plus larges de carbone organique et des bactéries indicatrices de contamination fécale. La microbiologie moléculaire (PCR-DGGE) a permis la caractérisation des communautés bactériennes de la perte et des sources. Ces résultats démontrent l'importante influence de la perte sur la qualité de l'eau souterraine, alors que sa contribution au débit du système est négligeable. Le carbone organique sembleêtre un meilleur indicateur de la présence de contamination bactérienne que la turbidité. RésuméResumen Se ha estudiado la dinámica del carbono orgánico, turbiedad, una bacteria indicadora de fecales, y parámetros fisicoquímicos en un sistema kárstico cerca de Yverdon, Suiza. Se realizaron mediciones en línea y muestreo en un sumidero que drena una superficie agrícola (la entrada), y dos grupos de manantiales (las salidas) que frecuentemente muestran contaminación bacterial. Un trazador fluorescente que se inyectó en el sumidero durante condiciones de flujo bajo arribó en los manantiales por vez primer...

The structure, diversity and dynamics of microbial communities from a swallow hole draining agricultural land and two connected karst springs (Switzerland) were studied using molecular microbiological methods and related to hydrological and physicochemical parameters. Storm responses and an annual hydrological cycle were monitored to determine the short-and long-term variability, respectively, of bacterial communities. Statistical analysis of bacterial genetic fingerprints (16S rDNA PCR-DGGE) of spring water samples revealed several clusters that corresponded well with different levels of the allochthonous swallow hole contribution. Microbial communities in spring water samples highly affected by the swallow hole showed low similarities among them, reflecting the high temporal variability of the bacterial communities infiltrating at the swallow hole. Conversely, high similarities among samples with low allochthonous contribution provided evidence for a stable autochthonous endokarst microbial community. Three spring samples, representative for low, medium and high swallow hole contribution, were analysed by cloning/sequencing in order to identify the major bacterial groups in the communities. The autochthonous endokarst microbial community was mainly characterized of δ-Proteobacteria, Acidobacteria and Nitrospira species. A high percentage of unknown sequences suggested further that many karst aquifer bacteria are still undiscovered. Finally, the potential use of groundwater biomonitoring using microbial communities is discussed.

Recharge and contamination of karst aquifers often occur via the unsaturated zone, but the functioning of this zone has not yet been fully understood. Therefore, irrigation and tracer experiments, along with monitoring of rainfall events, were used to examine water percolation and the transport of solutes, particles, and fecal bacteria between the land surface and a water outlet into a shallow cave. Monitored parameters included discharge, electrical conductivity, temperature, organic carbon, turbidity, particle-size distribution (PSD), fecal indicator bacteria, chloride, bromide, and uranine. Percolation following rainfall or irrigation can be subdivided into a lag phase (no response at the outlet), a piston-flow phase (release of epikarst storage water by pressure transfer), and a mixed-flow phase (increasing contribution of freshly infiltrated water), starting between 20 min and a few hours after the start of recharge event. Concerning particle and bacteria transport, results demonstrate that (1) a first turbidity signal occurs during increasing discharge due to remobilization of particles from fractures (pulse-through turbidity); (2) a second turbidity signal is caused by direct particle transfer from the soil (flow-through turbidity), often accompanied by high levels of fecal indicator bacteria, up to 17,000 Escherichia coli/100 mL; and (3) PSD allows differentiation between the two types of turbidity. A relative increase of fine particles (0.9 to 1.5 lm) coincides with microbial contamination. These findings help quantify water storage and percolation in the epikarst and better understand contaminant transport and attenuation. The use of PSD as ''early-warning parameter'' for microbial contamination in karst water is confirmed.

Continuous monitoring of particle-size distribution (PSD), total organic carbon (TOC), turbidity, discharge and physicochemical parameters, together with analyses of fecal indicator bacteria, particularly Escherichia coli, made it possible to better understand the processes governing pathogen transport in karst groundwater and to establish PSD as indicator for possible microbial contamination of drinking water from karst springs. In the study area near Yverdon-les-Bains, Switzerland, tracer tests proved connection between a sinking stream draining agricultural land and several springs, 4.8–6.3 km away. Tracing and monitoring results demonstrate that (i) suspended particles (turbidity) in the spring water either originate from remobilization of sediments inside the aquifer (autochthonous) or from the sinking stream and land surface (allochthonous); (ii) allochthonous turbidity coincides with increased E. coli and TOC levels; (iii) PSD makes it possible to distinguish the two types of turbidity; (iv) a relative increase of finer particles (0.9–10 µm) indicates allochthonous turbidity and thus possible fecal contamination. The method permits to optimize water treatment and identify periods when the spring water must be rejected. Findings from other test sites confirm the feasibility of this approach.