Stefan Woelfl

First results on the water chemistry, algae and trophic status of an Andean acidic lake system of volcanic origin in Patagonia (Lake Caviahue)

The acidic caldera lake Caviahue (Patagonia, Argentina) and its main tributaries were studied on two dates during September 1998. The main results are: The acidity of the Lake Caviahue (pH: 2.56, acidity: >5 mmol H+ l−1) is controlled by the extremely acidic Upper Rio Agrio (pH: 1.78, acidity: >20 mmol H+ l−1). The high sulphate contents of both the river and the lake can be attributed to sulphuric acid generated by the uptake of sulphurous gases in the crater lake of Copahue Volcano at approximately 2800 m a.s.l. The high concentrations of both Fe and trace metals (e.g. Cr, Ni, Zn) in Lake Caviahue originate from sulphur–acid interactions with the predominantly volcanic geology of the catchment area. The P-rich andesitic geology influences both the Upper and Lower Rio Agrio and Lake Caviahue. Both were found to have high phosphorus concentrations (300–500 μg P l−1) indicative of a high potential for eutrophication. The plankton community consisted of bacterioplankton, phytoplankton and rotifers. The phytoplankton was dominated by one green alga, Keratococcus raphidioides (>90% of total abundance) followed by a green sphaerical and Chlamydomonas sp. The total phytoplankton density was about 15 000 cells ml−1 in the upper 10 m of the water column. Rotifers were represented by one bdelloid species and their abundance was highly variable (360–4040 ind l−1) in the water columm. In the Upper and Lower Rio Agrio, the epilithic community was dominated by one chloroccocal species and two species of Ulothricales. According to trophic categories based on phytoplankton density and TP concentration, Lake Caviahue can be classified as mesotrophic/eutrophic. However, chlorophyll a concentrations observed were not in agreement with this state.

Sampling, preservation and quantification of biological samples from highly acidic environments (pH ≤3)

Standard methods for sampling, fixation and quantification of organisms in highly acid environments with pH values at or below 3 are presented. Some important problems are discussed, together with recommendations on how to handle the material. Examples of specific problems are included, especially the effects of fixatives on the cell dimensions of species of Chlamydomonas, Oxytricha and Actinophrys, and on cell numbers of Actinophrys. Mixed populations of heterotrophic protists should be preserved with Lugols solution, because other fixatives do not permit the recognition of heliozoans.

A review on the occurrence and taxonomy of heterotrophic protists in extreme acidic environments of pH values ≤ 3

The occurrence and some important taxonomic problems of heterotrophic protists from extremely acidic (pH ≤3) environments are briefly discussed. Almost all information on the occurrence of heterotrophic protists from extremely acidic environments is restricted to acid mine drainage (AMD) or streams influenced by AMD. Most of the information is provided for ciliates. Very little information is available on the occurrence of flagellates, rhizopods and heliozoa in this environment. Within the ciliates Urotricha, Vorticella and Oxytricha dominate in acidic mining lakes. Actinophrys sp. is the most important heliozoan in these environments. There are many taxonomic problems which are not solved so far. Live observations and taxonomic methods adapted to the extreme chemical matrix are necessary for correct identification.

The use of a portable total reflection X-ray fluorescence spectrometer for field investigation ☆

b ´ Abstract A newly developed, portable total reflection X-ray fluorescence (TXRF) spectrometer was tested during a field campaign on Chilean lakes and a German river in January 2002. The field measurements were compared with laboratory measurements carried out on a stationary instrument in the German laboratory. For method validation certified reference material (NIST SRM 1640 Trace elements in natural water) and water samples from different freshwater sources were analyzed with both techniques and evaluated statistically. Based on these preliminary results, it could be concluded that the portable TXRF is a useful technique for the quantitative elemental screening of freshwater samples during field campaigns. Future tests with biological samples (e.g. biofilms and zooplankton), and suspended matter will provide information about the suitability of the portable TXRF for these materials. 2003 Elsevier B.V. All rights reserved.

Cold plasma ashing improves the trace element detection of single Daphnia specimens by total reflection X-ray fluorescence spectrometry ☆

The recently developed dry method for the element determination of single freshwater microcrustacean specimens (Daphnia) using total reflection X-ray fluorescence (TXRF) spectrometry showed that inhomogeneities of the biological material on the glass carriers resulted in some cases in high background and hampered the detection of certain trace elements (e.g. Cr, Ni). The aim of this study was to test how inhomogeneities of the biological material can be reduced using cold plasma ashing (CPA) techniques. For that, single specimens of the microcrustacean Daphnia pulex prepared according to the dry method were measured by TXRF before and after CPA. To determine the efficiency of the removal of organic matrix, the background and signal-to-background relationship of 28 samples were analyzed. The results showed (1) a highly significant reduction of the background by CPA fluctuating between 26 and 46% (all elements) and (2) a significant increase of the signal-to-background relationship by the factor 1.5–2.5 (all elements) and a much better detection of Cr, Pb, As and Se. The element concentrations (with exception of Cr, Ni and Pb) after ashing were in the same range or slightly higher than that before ashing. No significant differences between the two treatments were observed for Mn, As, Pb, Se (November), Sr (November), Cr (March) and Pb (March). The element concentration of P, K, Ca, Cu, Zn, Cr (November), Fe and Rb were significantly higher after ashing. In general, they increased by 1.5–13.6% and were highest for Rb (March) and P (November). In contrast, the element concentration of Ni and Cr (only March) decreased significantly after ashing (Ni: 91.6–92.1%, Cr: 91.3%). We recommend the use of CPA for biological material in the microgram-range as a routine method for TXRF analysis, especially when trace elements in minute concentrations are of interest.

Trophic parameters and method validation in Lake Riñihue (North Patagonia: Chile) from 1978 through 1997

El lago Rinihue es el ultimo en una cadena de lagos oligotroficos, de origen glacial, en del sur Chile, y fue sujeto de investigaciones limnologicas por el Dr. Hugo Campos y colaboradores de la Universidad Austral de Chile, Valdivia, Chile, desde 1970. El Profesor Campos fallecio en terreno en 1998 y esta publicacion ahora presenta una revision de estos datos con comentarios acerca de la metodologia concerniente a parametros troficos de este lago. Se presentan datos de nutrientes inorganicos, clorofila a, fitobiomasa, productividad primaria y profundidad de Secchi obtenidos a traves de un periodo de 21 anos entre 1978 y 1997, con el objetivo de evaluar cambios potenciales de estos parametros a traves del tiempo. Se utilizaron pruebas no parametricas de Wilcoxon para determinar tendencias temporales y ellas revelaron que: (1) el fosforo total mostro un aumento significativo (TP: 2,7 mg P L-1 a 6,5 mg P L-1), (2) la profundidad de Secchi, el fosforo reactivo soluble y la productividad primaria no cambiaron significativamente, (3) la clorofila a cambio significativamente entre diferentes anos, pero no revela una clara tendencia temporal, (4) el nitrato mostro una tendencia a aumentar e incrementar las concentraciones durante el periodo a inicios de 1990, aunque estos datos se debieron sin duda a anomalias metodologicas, y (5) la fitobiomasa mostro valores significativamente mas altos durante el periodo de 1980. El aumento significativo en el fosforo total y la tendencia a aumentar del nitrato podria reflejar un aumento en el uso de praderas dentro de la cuenca de drenaje. Se detectaron una serie de anomalias en los datos historicos relacionadas a la metodologia los cuales se analizaron y discutieron dentro del contexto de este estudio.

Tracing dissolved organic matter (DOM) from land-based aquaculture systems in North Patagonian streams.

Chile is the second largest producer of salmonids worldwide. The first step in the production of salmonids takes place in land-based aquacultures. However, the effects of the discharge from these aquacultures on stream dissolved organic matter (DOM) content, molecular composition and degradability are unknown. The aim of this study was thus to investigate the inputs of anthropogenic DOM from land-based aquaculture to the predominantly pristine river systems of North Patagonia. We hypothesized, that i) DOM exported from land-based aquaculture mainly consists of protein-like fluorescence (tyrosine and tryptophan) released from fish feces and food remains, and that ii) this DOM is highly degradable and therefore rapidly turned-over within the receiving streams. In the North Patagonian region we conducted a screening of ten land-based aquacultures and an intensive sampling campaign for one aquaculture. This was combined with longitudinal transects and a degradation experiment in order to couple the composition of DOM exported from land-based aquacultures to its degradability in streams. We measured dissolved organic carbon (DOC) concentration by high-temperature catalytic oxidation and DOM composition by fluorescence spectroscopy and parallel factor analysis. In the effluent of the ten screened aquacultures and in the repeated sampling of one aquaculture, we consistently found an increase of DOC concentrations and a dominance of protein-like fluorescence. The protein-like fluorescence rapidly disappeared downstream of the aquacultures, and in the degradation experiment. 21% of the DOC export from the repeatedly sampled aquaculture resulted from food addition and 76% from fish production. We conclude that large amounts of degradable DOM are exported from land-based aquacultures. This probably has strong effects on the ecological structure and function of North Patagonian streams, and similarly affected streams worldwide.

A method for trace element determination of single Daphnia specimens using total reflection X-ray fluorescence spectrometry☆

Abstract Two new preparation techniques for total-reflection X-ray fluorescence (TXRF) element determination of single freshwater crustacean specimens (dry weight: 3–40 μg ind−1) have been developed and tested using Daphnia pulex from a deep, oligotrophic freshwater lake located in southern Chile. Dry method: Specimens were washed with 0.2 μm filtered lake water and frozen in liquid nitrogen. The freeze-dried Daphnia specimens were weighed using an ultra-fine microbalance and placed on quartz glass carriers for TXRF analysis. Wet method: Specimens were washed with 0.2 μm filtered lake water and placed on quartz glass carriers for TXRF analysis and dried in air. The dry weight was determined using the previously established body length–dry weight relationship. Method validation for both the dry and the wet preparation method in combination with TXRF spectrometry for the element determination in small single freshwater crustaceans showed that both methods can be used for routine investigations. There were no significant differences between the dry and the wet methods concerning the elements Ca, K, Fe, Zn, Br, P, Cu, but the determination of Mn, S and Sr revealed significant differences between the two methods. It seems that the dry method yields more precise results, but the wet method is easier to handle in the field when samples cannot be fixed with liquid nitrogen.

Use of null models to explain crustacean zooplankton assemblages in North Patagonian lakes with presence or absence of mixotrophic ciliates (38°S, Chile)

Zooplankton assemblages in Northern Patagonian lakes are characterized by a low species richness and a marked dominance of calanoid copepods. The aim of the present study was to do a null model analysis to explain the composition of the zooplankton assemblages in two North Patagonian lakes. The first aspect to which null model analysis was applied was species co-occurrence, and there the model indicates that the species associations found are not random. The second aspect was niche sharing, and that analysis denotes that the species do not share niches. The third aspect tested was size overlap, and the model there denotes that the species have no size overlap. The results obtained indicate that the sites are relatively homogeneous, which would explain the random effect in species co-ocurrence, whereas the absence of niche sharing is due to the presence of different energetic resources at both sites investigated, which would separate the niche of each species from those of other faunistic elements. Finally, the third application of the null model analysis demonstrates that there is no competition among the species reported upon herein.

Land-based salmon aquacultures change the quality and bacterial degradation of riverine dissolved organic matter

Aquacultures are of great economic importance worldwide but pollute pristine headwater streams, lakes, and estuaries. However, there are no in-depth studies of the consequences of aquacultures on dissolved organic matter (DOM) composition and structure. We performed a detailed molecular level characterization of aquaculture DOM quality and its bacterial degradation using four salmon aquacultures in Chile. Fluorescence measurements, ultrahigh-resolution mass spectrometry, and nuclear magnetic resonance spectroscopy of the DOM revealed specific and extensive molecular alterations caused by aquacultures. Aquacultures released large quantities of readily bioavailable metabolites (primarily carbohydrates and peptides/proteins, and lipids), causing the organic matter downstream of all the investigated aquacultures to deviate strongly from the highly processed, polydisperse and molecularly heterogeneous DOM found in pristine rivers. However, the upstream individual catchment DOM signatures remained distinguishable at the downstream sites. The benthic algal biovolume decreased and the bacterial biovolume and production increased downstream of the aquacultures, shifting stream ecosystems to a more heterotrophic state and thus impairing the ecosystem health. The bacterial DOM degradation rates explain the attenuation of aquaculture DOM within the subsequent stream reaches. This knowledge may aid the development of improved waste processing facilities and may help to define emission thresholds to protect sensitive stream ecosystems.