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Microbial communities in karst groundwater and their potential use for biomonitoring

2009, Pronk, Michiel, Goldscheider, Nicola, Zopfi, Jakob

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.

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Characterizing Water Circulation and Contaminant Transport in Lake Geneva Using Bacteriophage Tracer Experiments and Limnological Methods

2007, Goldscheider, Nicola, Haller, Laurence, Poté, John, Wildi, Walter, Zopfi, Jakob

Multi-tracer tests with three types of marine bacteriophages (H4/4, H6/1, and H40/1), together with various limnological methods, including physicochemical depth profiling, surface drifters, deep current measurements, and fecal indicator bacteria analyses, have been applied to characterize water circulation and pathogen transport in the Bay of Vidy (Lake Geneva, Switzerland). The experimental program was carried out twice, first in November 2005, when the lake was stratified, and a second time during holomixis in February 2006. The bacteriophages were injected at three points at different depths, where contaminated waters enter the lake, including the outlet pipe of a wastewater treatment plant, a river, and a stormwater outlet. Thereafter, water samples were collected in the lake at 2 m depth during a 48 h sampling campaign. The results demonstrate that (i) contaminated river water spreads rapidly in the bay; (ii) a well-developed thermocline is highly effective in preventing contamination from the depth to rise up to the surface; (iii) rapid vertical mixing and pathogen transport occur under thermally homogeneous conditions; and (iv) repeated multi-tracer tests with bacteriophages are a powerful technique to assess water circulation and contaminant transport in lakes where high dilution occurs.

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Origin and spatial–temporal distribution of faecal bacteria in a bay of Lake Geneva, Switzerland

2009, Poté, John, Goldscheider, Nicola, Haller, Laurence, Zopfi, Jakob, Khajehnouri, Fereidoun, Wildi, Walter

The origin and distribution of microbial contamination in Lake Geneva’s most polluted bay were assessed using faecal indicator bacteria (FIB). The lake is used as drinking water, for recreation and fishing. During 1 year, water samples were taken at 23 points in the bay and three contamination sources: a wastewater treatment plant (WWTP), a river and a storm water outlet. Analyses included Escherichia coli, enterococci (ENT), total coliforms (TC), and heterotrophic plate counts (HPC). E. coli input flux rates from the WWTP can reach 2.5 × 1010 CFU/s; those from the river are one to three orders of magnitude lower. Different pathogenic Salmonella serotypes were identified in water from these sources. FIB levels in the bay are highly variable. Results demonstrate that (1) the WWTP outlet at 30 m depth impacts near-surface water quality during holomixis in winter; (2) when the lake is stratified, the effluent water is generally trapped below the thermocline; (3) during major floods, upwelling across the thermocline may occur; (4) the river permanently contributes to contamination, mainly near the river mouth and during floods, when the storm water outlet contributes additionally; (5) the lowest FIB levels in the near-surface water occur during low-flow periods in the bathing season.

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Dynamics and interaction of organic carbon, turbidity and bacteria in a karst aquifer system

2006, Pronk, Michiel, Goldscheider, Nicola, Zopfi, Jakob

La dynamique du carbone organique, de la turbidité, des bactéries indicatrices de contamination fécale et d’autres paramètres physico-chimiques a été étudiée dans un système karstique proche de la ville d’Yverdon-les-Bains, Suisse. Des mesures en continu ainsi que des échantillonnages ont été effectués à une perte drainant une zone agricole (input), et à deux groupes sourciers (output) qui montrent fréquemment une contamination bactérienne. En période d’étiage, un essai de traçage à l’uranine a été réalisé depuis la perte. Le traceur est apparu aux sources 10–12 jours après l’injection; la masse de restitution totale a été de 29%. Des essais précédents, réalisés en hautes eaux, ont montré des temps de transit plus court. Suite à un événement 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 semble être un meilleur indicateur de la présence de contamination bactérienne que la turbidité., 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., 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 primera 10–12 días después de que fue inyectado; el ritmo total de recuperación fue de 29%. Las pruebas de trazadores realizadas con anterioridad bajo condiciones de flujo alto aportaron tiempos de viaje más cortos. Después de una tormenta fuerte se observó un pico de turbiedad primario en los manantiales. El pico coincide con el limbo ascendente del hidrograma indicando remobilización de partículas alóctonas provenientes del acuífero. Un pico de turbiedad secundario ocurre varios días más tarde sugiriendo el arribo de partículas alóctonas provenientes del sumidero. Se observaron simultáneamente picos más amplios de carbono orgánico y bacteria. La aplicación de métodos de microbiología molecular (PCR-DGGE) permitieron caracterizar las comunidades de bacteria en el sumidero y los manantiales. Los resultados demuestran que el sumidero es una fuente importante de contaminación de aguas subterráneas mientras que su contribución a la recarga del acuífero es insignificante. El carbono orgánico parece ser un mejor indicador de contaminación bacterial que la turbiedad.

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Percolation and Particle Transport in the Unsaturated Zone of a Karst Aquifer

2008, Pronk, Michiel, Goldscheider, Nicola, Zopfi, Jakob, Zwahlen, François

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 μm) 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.