Katrin Wendt-Potthoff

Phylogenetic Composition, Spatial Structure, and Dynamics of Lotic Bacterial Biofilms Investigated by Fluorescent in Situ Hybridization and Confocal Laser Scanning Microscopy

A bstractThe phylogenetic composition, three-dimensional structure and dynamics of bacterial communities in river biofilms generated in a rotating annular reactor system were studied by fluorescent in situ hybridization (FISH) and confocal laser scanning microscopy (CLSM). Biofilms grew on independently removable polycarbonate slides exposed in the reactor system with natural river water as inoculum and sole nutrient and carbon source. The microbial biofilm community developed from attached single cells and distinct microcolonies via a more confluent structure characterized by various filamentous bacteria to a mature biofilm rich in polymeric material with fewer cells on a per-area basis after 56 days. During the different stages of biofilm development, characteristic microcolonies and cell morphotypes could be identified as typical features of the investigated lotic biofilms. In situ analysis using a comprehensive suite of rRNA-targeted probes visualized individual cells within the alpha-, beta-, and gamma-Proteobacteria as well as the Cytophaga–Flavobacterium group as major parts of the attached community. The relative abundance of these major groups was determined by using digital image analysis to measure specific cell numbers as well as specific cell area after in situ probing. Within the lotic biofilm community, 87% of the whole bacterial cell area and 79% of the total cell counts hybridized with a Bacteria specific probe. During initial biofilm development, beta-Proteobacteria dominated the bacterial population. This was followed by a rapid increase of alpha-Proteobacteria and bacteria affiliated to the Cytophaga–Flavobacterium group. In mature biofilms, alpha-Proteobacteria and Cytophaga–Flavobacteria continued to be the prevalent bacterial groups. Beta-Proteobacteria constituted the morphologically most diverse group within the biofilm communities, and more narrow phylogenetic staining revealed the importance of distinct phylotypes within the beta1-Proteobacteria for the composition of the microbial community. The presence of sulfate-reducing bacteria affiliated to the Desulfovibrionaceae and Desulfobacteriaceae confirmed the range of metabolic potential within the lotic biofilms.

Occurrence and role of algae and fungi in acid mine drainage environment with special reference to metals and sulfate immobilization

Passive remediation of Acid Mine Drainage (AMD) is a popular technology under development in current research. Roles of algae and fungi, the natural residents of AMD and its attenuator are not emphasized adequately in the mine water research. Living symbiotically various species of algae and fungi effectively enrich the carbon sources that help to maintain the sulfate reducing bacterial (SRB) population in predominantly anaerobic environment. Algae produce anoxic zone for SRB action and help in biogenic alkalinity generation. While studies on algal population and actions are relatively available those on fungal population are limited. Fungi show capacity to absorb significant amount of metals in their cell wall, or by extracellular polysaccharide slime. This review tries to throw light on the roles of these two types of microorganisms and to document their activities in holistic form in the mine water environment. This work, inter alia, points out the potential and gap areas of likely future research before potential applications based on fungi and algae initiated AMD remediation can be made on sound understanding.

Biogeochemistry of Iron and Sulfur in Sediments of an Acidic Mining Lake in Lusatia, Germany

Chemical, microbiological and stable isotope analyses of sediments from an acidic mining lake were used to evaluate whether biogeochemical processes, such as iron and sulfate reduction, are extant, because such processes can potentially generate alkalinity. Sediment cores were sliced in cm intervals to achieve a high resolution for spatial distribution of organic and inorganic components. Iron, sulfur, carbon, nitrogen and phosphorus as well as the most probable number (MPN) of iron reducing bacteria, the amount of lipid phosphate and the stable isotope compositions of various sedimentary sulfur compounds were measured. Accumulation of degradable organic material, reduced mass fractions of iron, enhanced concentrations of lipid phosphate, high concentrations of DOC and ferrous iron in the pore water and a drastic change of sulfur isotope ratios in the upper 3 cm of the sediment all indicated a highly reactive zone of biogeochemical transformations. The data provide clear evidence for iron and sulfate reducing processes in the sediments that result in an increase of pH with depth.

Functional Groups and Activities of Bacteria in a Highly Acidic Volcanic Mountain Stream and Lake in Patagonia, Argentina

Acidic volcanic waters are naturally occurring extreme habitats that are subject of worldwide geochemical research but have been little investigated with respect to their biology. To fill this gap, the microbial ecology of a volcanic acidic river (pH approximately equal to 0-1.6), Rio Agrio, and the recipient lake Caviahue in Patagonia, Argentina, was studied. Water and sediment samples were investigated for Fe(II), Fe(III), methane, bacterial abundances, biomass, and activities (oxygen consumption, iron oxidation and reduction). The extremely acidic river showed a strong gradient of microbial life with increasing values downstream and few signs of life near the source. Only sulfide-oxidizing and fermentative bacteria could be cultured from the upper part of Rio Agrio. However, in the lower part of the system, microbial biomass and oxygen penetration and consumption in the sediment were comparable to non-extreme aquatic habitats. To characterize similarities and differences of chemically similar natural and man-made acidic waters, our findings were compared to those from acidic mining lakes in Germany. In the lower part of the river and the lake, numbers of iron and sulfur bacteria and total biomass in sediments were comparable to those known from acidic mining lakes. Bacterial abundance in water samples was also very similar for both types of acidic water (around 10(5) mL(-1)). In contrast, Fe(II) oxidation and Fe(III) reduction potentials appeared to be lower despite higher biogenic oxygen consumption and higher photosynthetic activity at the sediment-water interface. Surprisingly, methanogenesis was detected in the presence of high sulfate concentrations in the profundal sediment of Lake Caviahue. In addition to supplementing microbiological knowledge on acidic volcanic waters, our study provides a new view of these extreme sites in the general context of aquatic habitats.

Geostatistical Analysis of Surface Sediments in an Acidic Mining Lake

An exact morphometric description of Mining Lake 111, Brandenburg, Germany, was obtained after a new survey in 1996 by the staff of the water research department. Volume and surface area of ML111 were calculated based on values of this survey. An actual bathymetric map was created from the datapool which was the basis for the selection of sampling sites where we obtained sediment cores to describe the geochemistry of the lake sediments. Principal Component Analysis (PCA) was carried out to examine patterns and similarities between concentrations of different heavy metals. The patterns are different for the three basins within the lake. The spatial distributions of 12 elements in the sediment surface were estimated with a geostatistical procedure (Ordinary Kriging) as well as with a conventional interpolation method. The structure found with PCA was confirmed by the plots from spatial interpolation. Due to the complexity of the lake morphometry and bathymetry 66 sediment cores taken at 47 sampling sites were not sufficient to reduce the spatial variance distributions to acceptable values.

Microbial Fe(III) Reduction in Acidic Mining Lake Sediments after Addition of an Organic Substrate and Lime

To elucidate the role of Fe(III) reduction in mining lake sediments amended with organic substrates, we performed a large (10 m diameter) enclosure experiment in which sediments were amended with Carbokalk, a waste product from sugar industry containing organic carbon and lime. Fe(III) reduction rates were determined monthly by measuring the accumulation of Fe(II) in the sediments in the field. Fe(III) reduction rates were also determined by incubating sediment samples with synthetic Fe(III) oxyhydroxide under in situ temperature in the laboratory. Sulfate reduction was selectively inhibited in the Fe(III) reduction experiments by addition of sodium molybdate. Sulfate reduction was measured by accumulation of reduced inorganic sulfides in the field and by 35S radiotracer using a core injection technique. Sediment incubation and determination of sulfate reduction rates with radiotracer showed that sulfate reduction and direct microbial Fe(III) reduction occured simultaneously in the upper centimeters of the sediments and that both processes contributed to alkalinity generation. However, Fe(III) reduction was the initial process and rates were at least 3.5 fold higher than sulfate reduction rates. The results indicate that the presence of suitable anions for Fe(II) precipitation as carbonate or sulfide is needed in order to prevent loss of potential alkalinity by Fe(II) diffusion and reoxidation in the water column.

Reservoirs as sentinels of catchments: the Rappbode Reservoir Observatory (Harz Mountains, Germany)

Reservoirs can be viewed as sentinels of their catchments and a detailed monitoring of reservoir systems informs about biogeochemical and hydrological processes at the catchment scale. We developed a comprehensive online monitoring system at Rappbode reservoir, the largest drinking water reservoir in Germany, and its inflows. The Rappbode Reservoir Observatory comprises of a set of online-sensors for the measurement of physical, chemical, and biological variables and is complemented by a biweekly limnological sampling schedule. Measurement stations are deployed at the four major inflows into the system, at the outlets of all pre-reservoirs, as well as in the main reservoir. The newly installed monitoring system serves both scientific monitoring and process studies, as well as reservoir management. Particular emphasis is paid to the monitoring of short-term dynamics and many variables are measured at high temporal resolution. As an example, we quantitatively documented a flood event which mobilised high loads of dissolved organic carbon and changed the characteristics of the receiving reservoir from eutrophic to dystrophic within a few days. This event could have been completely missed by conventional biweekly sampling programs, but is relevant for biogeochemical fluxes at the catchment scale. We also show that the high frequency data provide a deeper insight into ecosystem dynamics and lake metabolism. The Rappbode Reservoir Observatory; moreover, offers a unique study site to apply, validate, and develop state of the art lake models to improve their predictive capabilities.

Functions of Straw for In Situ Remediation of Acidic Mining Lakes

The addition of straw in combination with ‘Carbokalk’, a by-product from the sugar-industry, was successfully used to stimulate microbial alkalinity generation in an acidic mining lake. To get detailed information about functions of straw, anenclosure experiment was carried out. Straw bundles were placedat the sediment surface of an acidic mining lake (ML 111) and thephysiochemical conditions and the microbiology of the sediment-water contact zone were studied. Straw was degraded by anaerobic microorganisms and dissolved organic carbon (DOC) leached from straw bundles. Pigmented flagellates responded to the DOC supply in the water column anda considerable amount of algal carbon was transported to the sediment. Straw addition led to microbial reduction of iron andsulfate in the sediment. Sulfate reduction was observed at a pHof 5.5. The pH, however, was not high enough to precipitate H2S completely. Thus, some H2S diffused into the watercolumn, where it was reoxidized. Straw did not create orstabilize an anoxic water body above the sediment. Microbial sulfate reduction and pyrite formation only took place in the sediment,whereas iron reduction also took place in the straw. Straw, however, altered the flow conditions above the sediment surfaceand prevented complete mixing of the profundal water. Straw didnot serve as a substratum for a reactive biofilm. We conclude that the most important function of straw for mining lake remediation is to be a long-term nutrient source for microbialalkalinity generation in the sediment.

Structure and function of the microbial community in an in situ reactor to treat an acidic mine pit lake

Sulfate-reducing bioreactors are a promising option for the treatment of acid mine drainage. We studied the structure and function of a biofilm in a methanol-fed fixed-bed in-lake reactor for the treatment of an acidic pit lake by a combination of laboratory incubations, chemical and molecular analyses and confocal laser scanning microscopy to determine whether competition by different groups of microorganisms as well as the precipitation of minerals affect reactor performance negatively. The biofilm growing on the surface of a synthetic carrier material consisted of dense microbial colonies covered by iron-sulfide precipitates. The microorganisms continuously had to overgrow this mineral coating, resulting in a high biomass turnover. About one third of the added methanol was used by sulfate reduction, and the rest by competing reactions. Sulfate-reducing bacteria as well as methanogens and acetogens were involved in methanol consumption. Six different groups of Deltaproteobacteria, dominated by the genera Desulfomonile, Desulfobacterium and a phylotype related to Geobacter, Gram-positive sulfate reducers of the genus Desulfosporosinus, acetogenic Acetobacteria, different fermenting bacteria as well as methylotrophic methanogens were identified. The versatility of the microbial food web is probably an important factor stabilizing the biofilm function under fluctuating and partly oxidizing conditions in the reactor.

Determination of heavy metals in biofilms from the River Elbe by total-reflection X-ray fluorescence spectrometry

Abstract Heavy metal contents of aquatic biofilms isolated from stones collected from, and ceramic plates exposed in, the River Elbe were determined by total-reflection X-ray fluorescence spectrometry (TXRF). The fractions of several elements (K, Ca, Cr, Mn, Fe, Ni, Cu, Zn, Pb) referred to the dry mass are 100 to 60 000-fold higher in the biofilm matrix compared with the bulk water phase. Biofilms grown on the exposed plates have a mass fraction of the determined elements higher by a factor of 2–3 compared with the biofilms derived from stones. These differences may be attributed to the different ages of the biofilms.