Development and composition of the epixylic biofilm in a blackwater river

1. Comparisons of chlorophyll a, bacterial density, frequencies of dividing cells, ash-free dry mass (AFDM) and extracellular polysaccharide content were made for biofilm developing on wood (Salix) submerged in replicated stream-side flumes exposed to either ambient light (light treatment) or covered to exclude light (dark treatment). Biofilm was sampled on days 3, 6, 9 and 14 during experimental periods occurring irrMay, September, November and December. 2. There were no significant differences in bacterial cell densities, frequencies of dividing cells, AFDM or extracellular poiysaccharide content between light and dark treatments. Ash content and bacterial biomass was similar to seston, suggesting the importance of seston as a source of material accumulating in the biofilm. 3. Of total epixylic organic carbon 7.2% was estimated to be extracellular polysaccharide, and 0.8% was bacterial carbon. At least nine times more carbon was contained in extracellular polysaccharide than in bacterial biomass. 4. In the epixylon of the Ogeechee River, bacterial dynamics appear to be controlled by factors other than the availability of algal substrates.     

Nutrient limitation of epilithic and epixylic biofilms in ten North American streams

1. Nutrient diffusing substrata were used to determine the effect of added inorganic nitrogen (N) and phosphorus (P) on the development of epilithic and epixylic biofilms in 10 North American streams. Four treatments of diffusing substrata were used: Control (agar only), N addition (0.5 m NaNO₃), P addition (0.5 m KH₂PO₄), and N + P combined (0.5 m NaNO₃ + 0.5 m KH₂PO₄). Agar surfaces were covered with glass fibre filters (for epilithon) or discs of untreated white oak wood veneer (for epixylon). 2. We found that if algae showed significant response to nutrient addition, N limitation (either N alone or N with P) was the most frequent response both on GF/F filters and on wood. Despite the low dissolved nutrient concentrations in our study streams, more than a third of the streams did not show any response to N or P addition. In fact, P was never the sole limiting nutrient for algal biofilms in this study. 3. Nutrient addition influenced algal colonisation of inorganic versus organic substrata in different ways. The presence of other biofilm constituents (e.g. fungi or bacteria) may influence whether algal biomass on wood increased in response to nutrient addition. Algae on organic and inorganic substrata responded similarly to nutrient addition in only one stream. 4. Fungal biomass on wood was nutrient limited in six of 10 study streams. N limitation of fungal biomass (with or without secondary P limitation) was most frequent, but P limitation did occur in two streams. 5. Our results show that biomass responses to nutrient addition by the heterotrophic and autotrophic components of the epixylic biofilm were different, though both experienced the same stream nutrient conditions. For algae and fungi growing on wood, limiting nutrients were rarely similar. Only three of nine streams showed the same biomass response to nutrient addition, including two that showed no significant change in biomass despite added nutrients.     

Spatial and temporal variability in epilithic biofilm bacterial communities along an upland river gradient

Riverine biofilms remain one of the least-studied habitats despite the significant increase in the examination of aquatic microbial communities in recent years. In this study, the dynamics of epilithic biofilm communities native on rocks from a low-order upland stream were examined over a period of 3 years. Spatial and temporal variations in bacterial communities were assessed using terminal restriction fragment length polymorphism, based on analysis of the 16S rRNA gene. In total, 108 epilithic biofilm samples were analysed and 170 different ribotypes were detected. A strong temporal gradient in ribotype composition was noticed, especially between samples collected in 2001 and those collected in 2002 and 2003, most likely reflecting interannual differences in weather conditions, such as temperature. A spatial gradient in ribotype composition, from upstream sites to the low-lying sites, was also evident and interpreted as an environmental variation gradient along the river course. Distinct biofilm communities consistently occurred at the first site along the river, which was significantly correlated to low pH. Temporal factors explained the highest degree of variation within the epilithic biofilms. Recurrent blooms of certain bacteria were noted within the system. Phylogenetic relationships of bacteria at one point in the river were determined using a cloning and sequencing approach, with Alphaproteobacteria dominating the community, followed by Cyanobacteria, Bacteroidetes and Betaproteobacteria.     

Biofilm development on leaf and wood surfaces in a boreal river

SUMMARY 1. Biofilms are organic layers that develop on submerged surfaces. They are composed of micro-organisms, exoenzymes, and detritus particles enclosed within a gelatinous matrix. While much is known about mineral surface biofilms, those developing on organic surfaces have not been extensively studied. We examined the influences of current velocity and substratum composition on biofilm development in a fourth-order North American boreal river. 2. Arrays of white birch ice-cream sticks and sugar maple leaves were placed at fast and slow current sites. Samples were collected periodically, analysed for mass loss, and assayed for microbial biomass (ATP, ergosterol, chlorophyll a) and exoenzyme activity associated with lignocellulose degradation (exo- and endocellulase, β-glucosidase, phenol oxidase, peroxidase). 3. Biofilms developed rapidly on both surfaces. On leaves, biomass peaked within 30 days of exposure. On wood, ATP and chlorophyll a concentrations peaked within 30–70 days, whereas ergosterol increased throughout the study (161 days). On leaves, current velocity had little influence on biofilm development, although breakdown rates were greater at the fast flow site. On wood, ATP and chlorophyll a concentrations were greater at the fast flow site, whereas ergosterol concentrations and breakdown rates were similar at both sites. Microbial biomass was consistently greater on wood than leaves, Exoenzyme activity developed rapidly on both surfaces; current velocity had little influence on activity. Except for β-glucosidase, activities were greater on wood than leaves. 4. Our results suggest that fungi are an important structuring element of organic surface biofilms and the physical stability of the substratum strongly influences biofilm development. Leaf surfaces are susceptible to softening and fragmentation, truncating biofilm development. In contrast, abrasion of wood surfaces removes senescent material exposing fresh substratum for colonization. Thus, wood surfaces with their greater physical stability, permit the development of more extensive biofilms. Wood surfaces may represent an overlooked but important site of metabolic activity in streams.     

Nutritional relationships among microorganisms in an epilithic biofilm community

Previous studies of an epilithic algal-bacterial community in a pristine mountain stream suggested that heterotrophic bacteria were responding to the metabolic activities of the phototrophic population. Subsequent studies were performed to follow the flow of labeled carbon, from its initial inorganic form, through the trophic levels of the mat community. A majority of primary production metabolites were excreted by the algal population during active growth; this shifted to an incorporation into cellular material as phototrophic activity declined. Results suggest that there was a direct flux of soluble algal products to the bacterial population, with little heterotrophic utilization of dissolved organics from the overlying stream water. Both phototrophic productivity and bacterial utilization of algal products peaked at approximately the same time of year. Activity of the diatom-dominated algal population declined as silica concentrations in the stream water dropped, leading to a situation in which the sessile bacteria were substrate limited. These events resulted in an almost complete disappearance of the community in early September.     

Spatial patchiness of epilithic biofilm caused by refuge-inhabiting high shore gastropods

Aggregations of Melarhaphe neritoides and Littorina saxatilis are common on the high shore in north-east England. These aggregations are frequently found in crevices which act as a refuge from desiccation. The aggregations are surrounded by patches of epilithic biofilm which are visibly lighter in colour than the biofilm present on the remainder of the high shore. Chlorophyll a levels were lower in regions of close proximity to aggregated littorinids (mean = 4.9 μg cm⁻²) as compared to areas >‰5 cm away from the edge of the visible colour change (mean = 15.9 μg cm⁻²). Manipulations of littorinid density showed that areas of high density had significantly lower levels of chlorophyll a than those where littorinids were excluded. This difference was mainly due to the presence of higher numbers of small grazed areas or an increase in size of grazed areas rather than a homogenous change in chlorophyll a levels across the entire manipulated area. These results are supported by observations showing that only 7% of the total foraging time of littorinids was spent outside visibly lighter patches of biofilm suggesting littorinids only control local biofilm levels through grazing.This paper has not been submitted elsewhere in identical or similar form, nor will it be during the first three months after its submission to Hydrobiologia     

Winter sites and cold-hardiness of two gastropod species in a boreal river

Summary The overwintering sites and cold-hardiness of Gyraulus acronicus and Valvata piscinalis were investigated in a northern Swedish river subject to wide seasonal fluctuations in water-level and ice-cover. More than 95% of the population of young and older G. acronicus overwintered in the frozen hydrolittoral zone. No specimens of V. piscinalis were found in the frozen zone. The cold-hardiness in G. acronicus in unfrozen sites was maximal in November and early December. V. piscinalis was not cold-hardy at any time of the year. These interspecific differences in cold-hardiness accord with the differences in overwintering habitats.     

Temporal and geographical distributions of epilithic sodium dodecyl sulfate-degrading bacteria in a polluted South Wales river

Epilithic bacteria were isolated nonselectively from riverbed stones and examined by gel zymography for their ability to produce alkylsulfatase (AS) enzymes and thus to metabolize alkyl sulfate surfactants such as sodium dodecyl sulfate. The percentages of AS+ isolates from stone epilithon at five sites from the source to the river mouth were measured on five sampling days spread over 1 year. The results showed that (i) the prevalence of epilithic AS+ strains (as a percentage of all isolates) was much higher at polluted sites than at the source; (ii) when averaged over the whole river, percentages of AS+ strains were significantly higher at the end of summer compared with either the preceding or the following winter; (iii) analysis of site-sampling time interactions indicated that water quality factors (e.g., biochemical oxygen demand and dissolved oxygen concentration) rather than climatic factors determined the distributions of epilithic AS+ isolates; (iv) constitutive strains were the most prevalent (7.2% of all isolates), with smaller numbers of isolates with inducible (4.5%) and repressible (1.7%) enzymes.     

Enzymic and chemical analysis of particulate organic matter from a boreal river

SUMMARY. 1. Benthic particulate organic matter (POM) was collected from a shallow pool of a fourth order boreal stream and sorted into seven size fractions ranging from 63 to >4000μm. Each size fraction was analysed for fibre, total phosphorus, and total Kjeldahl nitrogen. Microbial activity was measured by oxygen consumption and characterized by assaying for eleven classes of exoenzymes including cellulase, phenol oxidase, peroxidase, phosphatase and sulphatase.
2. Indices of detritus quality such as C/N, C/P, percent lignin, and microbial respiration showed improvement with decreasing particle size. Three covarying exoenzyme groups were identified: a carbohydrase-phosphatase group that included eight enzymes, a phenol oxidase-peroxidase group, and sulphatase. The activity of the carbohydrase-phosphatase group, was significantly correlated with microbial respiration and the carbohydrate content of the POM. Phenol oxidase-peroxidase activity was correlated with lignin content for POM greater than 250 μm, but activity increased markedly in the two smallest size fractions even though the lignin content of the POM continued to decline, Sulphatase activity was inversely related to particle size over the entire range.
3. The changes in microbial activity with particle size were attributed to the increasing surface area to volume ratio of smaller particles and to an ecological succession in the microbial community.     

Temporal and geographical distributions of epilithic sodium dodecyl sulfate-degrading bacteria in a polluted South Wales river.

Epilithic bacteria were isolated nonselectively from riverbed stones and examined by gel zymography for their ability to produce alkylsulfatase (AS) enzymes and thus to metabolize alkyl sulfate surfactants such as sodium dodecyl sulfate. The percentages of AS+ isolates from stone epilithon at five sites from the source to the river mouth were measured on five sampling days spread over 1 year. The results showed that (i) the prevalence of epilithic AS+ strains (as a percentage of all isolates) was much higher at polluted sites than at the source; (ii) when averaged over the whole river, percentages of AS+ strains were significantly higher at the end of summer compared with either the preceding or the following winter; (iii) analysis of site-sampling time interactions indicated that water quality factors (e.g., biochemical oxygen demand and dissolved oxygen concentration) rather than climatic factors determined the distributions of epilithic AS+ isolates; (iv) constitutive strains were the most prevalent (7.2% of all isolates), with smaller numbers of isolates with inducible (4.5%) and repressible (1.7%) enzymes.     

Seasonal variation in the lateral distribution of mayfly nymphs in a boreal river

Seasonal variations in the lateral distribution of mayfly nymphs were investigated in the North Swedish river Vindelälven; a river with large seasonal differences in water level, current velocity and ice conditions, A total of 22 mayfly species about equally divided between summer and winter species were found along a single transect. At low water level in late summer and autumn most species were distributed over the whole transect. In winter the populations occupying the littoral moved into deeper water, so avoiding the ice. Only very small mortalities were detected in the ice. Prior to the spring flood most characteristically still water species moved to the shallow uppermost littoral while lotic species and the burrowing Ephemera vulgata L. did not show the same aggregation near the shore. It is suggested that these lateral movements allow the nymphs to avoid adverse environmental conditions and to exploit food resources over the whole transect.     

Soft X-ray spectromicroscopy of nickel sorption in a natural river biofilm

Scanning transmission X-ray microscopy (STXM) at the C 1s, O 1s, Ni 2p, Ca 2p, Mn 2p, Fe 2p, Mg 1s, Al 1s and Si 1s edges was used to study Ni sorption in a complex natural river biofilm. The 10-week grown river biofilm was exposed to 10 mg L⁻¹ Ni²⁺ (as NiCl₂) for 24 h. The region of the biofilm examined was dominated by filamentous structures, which were interpreted as the discarded sheaths of filamentous bacteria, as well as a sparse distribution of rod-shaped bacteria. The region also contained discrete particles with spectra similar to those of muscovite, SiO₂ and CaCO₃. The Ni(II) ions were selectively adsorbed by the sheaths of the filamentous bacteria. The sheaths were observed to be metal rich with significant amounts of Ca, Fe and Mn, along with the Ni. In addition, the sheaths had a large silicate content but little organic material. The metal content of the rod-shaped bacterial cells was much lower. The Fe on the sheath was mainly in the Fe(III) oxidation state. Mn was found in II, III and IV oxidation states. The Ni was likely sorbed to Mn–Fe minerals on the sheath. These STXM results have probed nano-scale biogeochemistry associated with bacterial species in a complex, natural biofilm community. They have implications for selective Ni contamination of the food chain and for developing bioremediation strategies.     

Temporal variation in the drift of plant litter and propagules in a small boreal river

SUMMARY 1. Rivers are linear ecosystems across landscapes with an effective transport of organisms, sediment and organic matter. Dispersal is studied mostly during single events and for single species, and there is little knowledge on how the drift of plant litter and propagules varies within and between years for entire communities.
2. We used floating traps for collecting waterborne plant litter and propagules in a small boreal river over 2 years. We installed the traps at four different locations along the river, and emptied them at least once a week during the ice-free season. We analysed propagule content by sorting and identifying species and through germination tests on bare soil.
3. In total, we recorded at least 54 taxa in the samples, and the highest density recorded in one sample was 5000 propagules per 100 g litter (dry weight). Large temporal variations in litter and propagule transport were revealed, both within and between years.
4. The longitudinal pattern was consistent between years, with an increasing mass of litter and number of propagule taxa downstream. The results highlight the importance of the temporal and longitudinal dimensions in river management.     

Sorption and metabolism of selected herbicides in river biofilm communities

In the present study, biofilms were grown in rotating annular bioreactors with river water as inoculum and sole source of nutrients. The herbicides atrazine and diclofop methyl were applied to the bioreactors, while an identical reactor acted as a control. Biofilm structure was visualized using specific fluorescent probes in conjunction with confocal laser scanning microscopy. The concentration of both herbicides in the bulk water phase followed the pattern of application. Atrazine and metabolites were detected in biofilm samples using direct insertion probe tandem mass spectrometry (DIP-MS/MS) and only trace levels were detected after the addition phase. Monoclonal antibody (MAb) studies indicated that sorption of atrazine was associated with a unique microcolony type. In contrast, diclofop and metabolites reached a maximum level in the biofilm at the end of the addition phase and persisted in the biofilm. Experiments with 14C-labeled atrazine and diclofop methyl indicated that mineralization of these compounds to CO2 (<1%) occurred in the river biofilms. Thus, both herbicides were sorbed and metabolized by the river biofilm community and detected in biofilms when they were not detected in the bulk water phase. These results indicate that biofilms and specific community members may act as a sink for herbicides, and that this should be taken into account in terms of both sampling and studies of the environmental chemodynamics of contaminants.     

Assessing the importance of a self-generated detachment process in river biofilm models

1. Epilithic biofilm biomass was measured for 14 months in two sites, located up‐ and downstream of the city of Toulouse in the Garonne River (south‐west France). Periodical sampling provided a biomass data set to compare with simulations from the model of Uehlinger, Bürher and Reichert (1996: Freshwater Biology, 36 , 249–263.), in order to evaluate the impact of hydraulic disturbance. 2. Despite differences in application conditions (e.g. river size, discharge, frequency of disturbance), the base equation satisfactorily predicted biomass between low and high water periods of the year, suggesting that the flood disturbance regime may be considered a universal mechanism controlling periphyton biomass. 3. However modelling gave no agreement with biomass dynamics during the 7‐month long low water period that the river experienced. The influence of other biomass‐regulating factors (temperature, light and soluble reactive phosphorus) on temporal biomass dynamics was weak. 4. Implementing a supplementary mechanism corresponding to a temperature‐dependent self‐generated loss because of heterotrophic processes allowed us to accurately reproduce the observed pattern: a succession of two peaks. This case study suggests that during typical summer low water periods (flow stability and favourable temperature) river biofilm modelling requires self‐generated detachment to be considered.     

Changes in distribution and behaviour of two congeneric mayflies in a boreal river and its seasonal tributaries

Both Parameletus chelifer and P. minor had univoltine life-cycles. For a period of 6–10 months almost no nymphs of either species were found. The first nymphs appeared near midstream in winter and later in spring when the ice had broken up, they were found in great numbers close to the bank of the main river. During spring, nymphs of P. chelifer colonized the newly flooded littoral faster, and a seasonal stream in an alluvial meadow in greater numbers, than did nymphs of P. minor. Both species also colonized 10 seasonal tributaries which showed that the springtime upstream colonization was a common behaviour. During the colonization of the seasonal stream in the alluvial meadow, the diel activity pattern changed in nymphs of both species. In the main river small nymphs showed a nocturnal activity, while large nymphs were active during day-time. In the seasonal stream, both small and large nymphs showed a diurnal activity. However, when the stream was flooded, small nymphs of both species showed nocturnal activity again. In both species, diurnal activity of mature nymphs continued during emergence. Reasons for shifts in diel activity are discussed.     

Intermediate disturbance hypothesis explains eutrophication and biodiversity pattern in a boreal river basin,China

Hydrobiologia - Eutrophication is a common phenomenon in aquatic ecosystems caused by human disturbance and is regarded as one of the major threats to biodiversity in rivers throughout the world....     

Small scale application and assessment of a Index of Biotic Integrity for a large boreal river

Modifications and use of Karr’s Index of Biotic Integrity (IBI) for assessing the effects of anthropogenic impacts to aquatic ecosystems have typically occurred using data collected at the macro scale. However, some non-point sources of habitat degradation occur at small scales. One possible source of perturbation to fish habitat in boreal rivers is the application of rip rap shoreline armouring in human use areas. In this study we assess the use of IBI in a small scale setting and discuss the potential impact of rip rap shoreline armouring. We captured small and juvenile fishes weekly during 2002–2003 from 12 sample sites within the littoral zone of a human use area using a modified beach seine. Principal component analysis (PCA) was performed on the data to examine the relationship between species composition and IBI scores. We also performed PCA on the IBI metrics to assess our modifications. The IBI method produced higher scores for armoured sites than for unarmoured sites. We found a strong rank order correlation (Spearman’s ρ > 0.93; p < 0.001) between the modified IBI scores and the first principal component, suggesting that Karr’s original empirically-based IBI is strongly linked to species composition. We found a high degree of redundancy between the metrics of the IBI that validate our modifications. These results suggest that IBI can be a suitable method for assessing non-point impacts from within a small study area. Our results also indicate that fish habitat from rip rap armoured sites within the test area had consistently higher IBI scores that unarmoured sites.     

Genetic diversity in populations of plants with different breeding and dispersal strategies in a free-flowing boreal river system

We have studied the genetic diversity of three plant species: Angelica archangelica (Apiacieae), Bistorta vivipara (Polygonaceae) and Viscaria alpina (Caryophyllaceae) along the free-flowing Vindel River in northern Sweden. The plants differ in reproductive strategy. A archangelica and V. alpina are insect pollinated outbreeders while B. vivipara reproduces with apomixis through bulbils. The seeds of A. archangelica may float for over a year, while the propagules (seeds and bulbils, respectively) of V. alpina and B. vivipara float for less than two days. Genetic diversity was assessed using starch gel electrophoresis. The clonal diversity of B. vivipara measured by Simpson's index (D) ranged between 0.78 and 0.99. Only a few clones were shared between localities. The average percentages polymorphic loci and mean He based on polymorphic loci for V. alpina over all localities were 23.1 and 0.15, respectively. Wright's F-statistics showed a significant overall deficit of heterozygotes. The diversity of A. archangelica was found to increase downstream. Genetic diversity of each species is sufficiently high to be used in studies on hydrochory. Dispersal appears to be related to the floating ability of progagules.