Influence of Algal Biomass on Extracellular Enzyme Activity in River Biofilms

Abstract Relationships between biofilm structural components (algal and bacterial biomass) and the activities of some extracellular enzymes that contribute to the ability to degrade organic matter) were explored for six Atlantic and three Mediterranean streams and rivers. The biofilms in these fluvial systems accounted for a wide range of bacterial and algal biomass and colonized the most common benthic habitats. Ratio of bacteria/algae biomass was lower in Atlantic than in Mediterranean streams, but enzymatic activities (β-glucosidase, β-xylosidase, phosphatase) were in general greater in the Mediterranean stream biofilms. Climatic characteristics (especially temperature) may explain the differences in enzymatic activities between biofilms of similar structure but different flow regime. The ratio β-xylosidase: β-glucosidase was similar (around 0.5) for all streams and substrata considered, showing that there is a general higher utilization of cellobiosic than xylobiosic molecules in fluvial systems. In general, highly heterotrophic biofilms showed lower extracellular enzymatic activities than more autotrophic biofilms. Maximum enzymatic activity is achieved when the algal biomass is two- to threefold higher than the bacterial biomass. The relevance of algal biomass on the heterotrophic ability of biofilms may be related to the physical proximity between the two, but also to the high proportion of polymeric carbohydrates included in algal exudates and lysis products, whose use is enzyme-mediated. Received: January 2000; Accepted: April 2000; Online Publication: 18 July 2000     

Nutrient dynamics and successional changes in a lentic freshwater biofilm

SUMMARY 1. Colonisation, species composition, succession of microalgae and nutrient dynamics in biofilms grown under light and dark conditions were examined during the initial phases of biofilm development in a lentic freshwater environment.
2. Biofilms were developed on inert (perspex) panels under natural illuminated and experimental dark conditions and the panels were retrieved for analysis after different incubation periods. Analysed parameters included biofilm thickness, algal density, biomass, chlorophyll a , species composition, total bacterial density and nutrients such as nitrite, nitrate, phosphate and silicate.
3. Biofilm thickness, algal density, biomass, chlorophyll a and species richness were significantly higher in light-grown biofilms, compared with dark-grown biofilms. The light-grown biofilms showed a three-phased succession pattern, with an initial domination of Chlorophyceae followed by diatoms (Bacillariophyceae) and finally by cyanobacteria. Dark-grown biofilms were mostly dominated by diatoms.
4. Nutrients were invariably more concentrated in biofilms than in ambient water. Nutrient concentrations were generally higher in dark-grown biofilms except in the case of phosphate, which was more concentrated in light-grown biofilms. Significant correlations between nutrients and biofilm parameters were observed only in light-grown biofilms.
5. The N : P ratio in the biofilm matrix decreased sharply in the initial 4 days of biofilm growth; ensuing N-limitation status seemed to influence biofilm community structure. The N : P ratios showed significant positive correlations with the chlorophycean fraction in both light and dark-grown biofilms, and low N : P ratio in the older biofilms favoured cyanobacteria. Our data indicate that nutrient chemistry of biofilm matrix shapes community structure in microalgal biofilms.     

Integrated abundance and biomass of sympagic meiofauna in Arctic and Antarctic pack ice

The abundance and biomass of sympagic meiofauna were studied during three cruises to the Antarctic and one summer expedition to the central Arctic Ocean. Ice samples were collected by ice coring and algal pigment concentrations and meiofauna abundances were determined for entire cores. Median meiofauna abundances for the expeditions ranged from 4.4 to 139.5 × 10³ organisms m⁻² in Antarctic sea ice and accounted for 40.6 × 10³ organisms m⁻² in Arctic multi-year sea ice. While most taxa (ciliates, foraminifers, turbellarians, crustaceans) were common in both Arctic and Antarctic sea ice, nematodes and rotifers occurred only in the Arctic. Based on the calculated biomass, the potential meiofauna ingestion rates were determined by applying an allometric model. For both hemispheres, daily and yearly potential ingestion rates were below the production values of the ice algal communities, pointing towards non-limited feeding conditions for ice meiofauna year-round. Accepted: 29 March 1999     

Bacteria and algae in stream periphyton along a nutrient gradient

1. Stream riffles in southern Ontario and western Quèbec were sampled for biomass (58 stations from 51 streams) and production (22 stations from 21 streams) of algae and bacteria in periphyton to test the hypothesis that bacteria in benthic biofilms compete with algae for nutrients. 2. Algal and bacterial biomass were positively correlated, as were algal and bacterial production. Bacterial production was also positively correlated to algal and bacterial biomass, but the relationship was not significant. The ratio of algal to bacterial biomass did not vary with nutrients whereas algal production tended to increase with nutrients more rapidly than bacterial production. 3. Instream nitrogen concentrations explained 38–58% of the variability in algal biomass and production. Bacterial abundance explained an additional 9–29% of the residual variance in algal production and biomass. However, the relationship between bacterial abundance and algal production and biomass, once nutrients were taken into account, was positive, in contrast to the predicted effect of competition. 4. Hence, we reject our original hypothesis that bacteria in biofilms compete with algae for nutrients and instead suggest that bacteria and algae in biofilms coexist in an association that offers space and resources to sustain production of both groups of organisms.     

Biofilm Structure and Function and Possible Implications for Riverine DOC Dynamics

Biofilms are major sites of carbon cycling in streams and rivers. Here we elucidate the relationship between biofilm structure and function and river DOC dynamics. Metabolism (extracellular enzymatic activity) and structure (algae, bacteria, C/N content) of light-grown (in an open channel) and dark-grown (in a dark pipe) biofilms were studied over a year, and variations in dissolved organic carbon (DOC) and biodegradable DOC (BDOC) were also recorded. A laboratory experiment on ¹⁴C-glucose uptake and DOC dynamics was also performed by incubating natural biofilms in microcosms. On the basis of our field (annual DOC budget) and laboratory results, we conclude that light-grown biofilm is, on annual average, a net DOC consumer. This biofilm showed a high monthly variability in DOC uptake/release rates, but, on average, the annual uptake rate was greater than that of the dark-grown biofilm. The higher algal biomass and greater structure of the light-grown biofilm may enhance the development of the bacterial community (bacterial biomass and activity) and microbial heterotrophic activity. In addition, the light-grown biofilm may promote abiotic adsorption because of the development of a polysaccharide matrix. In contrast, the dark-grown biofilm is highly dependent on the amount and quality of organic matter that enters the system and is more efficient in the uptake of labile molecules (higher ¹⁴C-glucose uptake rate per mgC). The positive relationships between the extracellular enzymatic activity of biofilm and DOC and BDOC content in flowing water indicate that biofilm metabolism contributes to DOC dynamics in fluvial systems. Our results show that short-term fluvial DOC dynamics is mainly due to the use and recycling of the more labile molecules. At the river ecosystem level, the potential surface area for biofilm formation and the quantity and quality of available organic carbon might determine the effects of biofilm function on DOC dynamics.     

Does biofilm origin matter? Biofilm responses to non-flow period in permanent and temporary streams

1. In some regions, climate change is increasing the variability of rainfall and the frequency of extreme events such as drought. Consequently, non-flow periods have grown in length and frequency, both in temporary and in formerly permanent streams. Water abstraction for human use may further prolong these dry periods.
2. We analysed the resistance and resilience of biofilms from permanent and temporary streams to non-flow conditions. This was achieved by exposing cobbles (collected from permanent and temporary streams) with intact biofilm to 31 days of non-flow, followed by 20 days of stream flow in artificial stream channels. Biofilm resistance and resilience were assessed at a structural (algal biomass, pigment composition, and algae and cyanobacteria composition) and functional level (photosynthetic efficiency and community metabolism).
3. Algal taxa in biofilms from permanent and temporary streams differed throughout the experiment. Biofilms from permanent streams were less resistant to non-flow than those from temporary streams at structural level. Permanent stream biofilms also presented lower resilience at a structural level, but responded similarly to temporary stream biofilms at a functional level.
4. Our investigation shows how the non-flow period disturbed permanent stream biofilms, and suggests that temporary stream biofilms will have greater adaptive capacity as hydroperiod becomes shorter due to climate change.

     

Microscale evaluation of the effects of grazing by invertebrates with contrasting feeding modes on river biofilm architecture and composition

River biofilms are a valuable food resource for many invertebrates. In the present study biofilms were cultivated in a rotating annular bioreactor with river water as sole source of inoculum. The resulting biofilms were then presented to starved snails, ostracods, and mayflies as sole food source. The biofilms were then removed and microscopically examined to determine areas that had been grazed. The grazed and ungrazed areas were marked and analyzed for the effects of grazing using confocal laser scanning microscopy and image analyses. Samples were treated with fluorescent probes for nucleic acids to quantify bacterial biomass and fluor-conjugated lectins to quantify exopolymer, and far red autofluorescence was imaged to quantify algal or photosynthetic biomass. Grazing by snails significantly reduced algal biomass (1.1 +/- 0.6 micro m 3 micro m 2 to 0.02 +/- 0.04 micro m 3 micro m 2), exopolymer (5.3 +/- 3.4 micro m 3 micro m 2 to 0.18 +/- 0.18 micro m 3 micro m 2), and biofilm thickness (154 micro m +/- 50 to 11 micro m +/- 5.2; ANOVA, p < or= 0.05). Although bacterial biomass was influenced by grazing snails the impact was not statistically significant (p 相似文献   

New penis characters to distinguish between two American Artemia species

Biofilms are an ensemble of autotrophs and heterotrophs, which are highly efficient in removing inorganic and organic compounds, as well as other chemicals, from river water. They are, therefore, key elements in the self-purification processes which occur in rivers. Biofilm function is related to several environmental factors that govern river ecosystems: physical (light, temperature, water current), chemical (nutrient availability, toxicant effects), but also biological. Among the biological factors, community composition (algae, bacteria and fungi), biofilm structure (layer arrangement and biomass accumulation), and the presence of grazers determine variations in the efficiency of the self-depuration function of biofilms in rivers. Algae and bacteria show specific abilities for nutrients and other organic and inorganic compounds, but biofilm thickness may affect these abilities, both through a decrease in diffusion and by enhancing recycling within the biofilm. Nutrient uptake and consequently the capacity of biofilm to ameliorate water quality decreases with biomass. Moreover, biofilm thickness determines the effect of toxicants, since biomass prevents their diffusion through the biofilm. Grazing interferes in the relative efficiency of biofilms, by simplifying the composition of the biofilm community and by decreasing the amount of sorption and uptake of the biofilm. Closer attention should be paid to these aspects, since they unambiguously interfere with the performance of biofilms in the amelioration of the quality of river water.     

Longitudinal distribution of macroinvertebrates in two glacier-fed New Zealand rivers

1. Environmental variables, benthic algal biomass and macroinvertebrate fauna were examined from September 1999 to January 2000 (austral summer) along two glacier-fed rivers in South Island, New Zealand.
2. The rivers were characterized by high flow variability, high turbidity and physically disturbed beds. Water temperature ranged from <1 °C near the glacier margin to 10 °C further downstream.
3. Epilithic algal biomass was very low (<0.1 mg m^–2) in months characterized by heavy rainfall, but ranged from 1.1 to 14.4 mg m^–2 following an extended period with negligible precipitation.
4. Abundance and diversity of invertebrates in both rivers was low. Dominant taxa were Chironomidae (Orthocladiinae, Podonominae, Diamesinae), although mayfly species ( Deleatidium : Leptophlebiidae) also occurred at most sites. A species of Eukiefferiella (Orthocladiinae) was collected at all sites and was the most abundant invertebrate close to the glacier margins. No meiofauna were found in either river.
5. Faunal diversity increased at the lowermost stations where species of Plecoptera, Trichoptera, Coleoptera and non-chironomid Diptera also occurred.
6. The faunas of the two New Zealand rivers conformed to the conceptual model of Milner & Petts (1994) in that taxon richness increased downstream with water temperature. However, invertebrate abundance increased downstream in only one of the two rivers. Also in contrast to the model predictions, Leptophlebiidae and Orthocladiinae, rather than Diamesinae, dominated the fauna at the coldest sites.     

Temporal Dynamics of River Biofilm in Constant Flows: A Case Study in a Riverside Laboratory Flume

A 15‐week experiment was performed in a riverside laboratory flume (with diverted river water) to check variations of river biofilm structure (biomass, algal and bacterial compositions) and function (community gross primary production GPP and respiration) under constant flow while water quality went through natural temporal variations. One major suspended matter pulse coinciding with one river flood was recorded after 10 weeks of experiment. Epilithic biofilm first exhibited a 10‐week typical pattern of biomass accrual reaching 33 g ash‐free dry matter (AFDM) m^–2 and 487 mg chlorophyll‐a m^–2 and then, experienced a shift to dominance of loss processes (loss of 60% AFDM and 80% chlorophyll‐a) coinciding with the main suspended matter pulse. Algal diversity remained low and constant during the experiment: Fragilaria capucina and Encyonema minutum always contribute over 80% of cell counts. DGGE banding patterns discriminated between two groups that corresponded to samples before and after biomass loss, indicating major changes in the bacterial community composition. GPP/R remained high during the experiment, suggesting that photoautotrophic metabolism prevailed and detachment was not autogenic (i.e., due to algal senescence or driven by heterotrophic processes within the biofilm). Observational results suggested that silt deposition into the biofilm matrix could have triggered biomass loss. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Effect of main-stem dams on zooplankton communities of the Missouri River (USA)

The persistence of plankton in flowing water presents an enigma, i.e., how can populations be sustained while constantly losing individuals downriver? We examined the distribution and abundance of zooplankton from 146 sites on the Missouri River (USA) and found large shifts in the dominance of major taxa between management zones of this regulated river. Crustacean zooplankton were dominant in the inter-reservoir zone of the river, and their taxonomic composition was similar to regional lakes and reservoirs. The exponential decline of cladocerans and copepods with distance from main-stem dams suggests that conditions within the river are adverse to population growth and that reservoirs are the main source of these crustaceans in the river. Rotifers dominated in the channelized zone of the river. High algal biomass and rapid population growth rates likely allow persistence of rotifers in segments of the river that do not receive direct reservoir inputs. Rotifers were less abundant in the inter-reservoir zone, suggesting that their numbers are limited by internal processes, such as food or predators. Since zooplankton are known to be an important food for larval fishes in rivers, this shift of major taxa in regulated rivers has implications for river food webs.     

Exoenzyme accumulation in epilithic biofilms

Although exoenzyme accumulation is often proposed as an explanation for the high metabolic activity of biofilms, little is known about the abundance, distribution and turnover rates of exoenzymes within these communities. To assess accumulation, epilithic biofilm samples were collected from a fourth-order boreal river and homogenized. The resulting particles were fractionated by size and each fraction was assayed for nine exoenzyme activities, chlorophyll, and ATP. In general, carbohydrase activities were not correlated with microbial biomass indicators; the largest pool of activity was in the aqueous phase (< 0.2 µm). Phenol oxidase, peroxidase, and phosphatase activities were largely particle-bound and often correlated with microbial biomass distribution. It was concluded that the epilithic biofilm matrix was effective at accumulating carbohydrase activity and that this accumulation may partially account for the metabolic resistance of epilithic biofilms to dissolved organic matter fluctuations.     

Seasonal determinations of extracellular hydrolytic activities in heterotrophic and mixed heterotrophic/autotrophic biofilms from two contrasting rivers

The temporal changes in extracellular enzyme activities in freshwater microbial biofilms were examined in two contrasting river sites in North Wales over a 12 month period. Sites were a first order, unshaded oligotrophic upland stream (Nant Waen) and a fourth order, mildly eutrophic river with riparian tree cover (River Clywedog). When algal populations were low, biofilms of the more eutrophic site supported greater enzyme activities and higher population densities than the oligotrophic site. Composition, concentration and origin of substrates available to the respective biofilm communities influenced extracellular processing patterns. Reduction in algal populations depressed total and extracellular activities in biofilms from the first order site, suggesting that biofilm communities here were maintained by in situ primary production. Biofilms from Nant Waen were often found to contain higher extracellular activities per cell than the more eutrophic River Clywedog biofilms, which might represent the enhanced ability of an oligotrophic biofilm to accumulate extracellular enzymes. In contrast, light and darkgrown River Clywedog biofilms were not enzymatically distinct, inferring a less important role for biofilm phototrophs. Some evidence was found for increased reliance on allochthonous substrates in the River Clywedog for biofilm maintenance.     

Influence of Algal Photosynthesis on Biofilm Bacterial Production and Associated Glucosidase and Xylosidase Activities

Natural photosynthetic biofilms were incubated under light (100 mmol m-2 s-1) and dark conditions to elucidate the impact of photosynthesis on bacterial production, abundance, biovolume, biomass, and enzyme activities over 24 h. Use of organic carbon-free media limited carbon sources to algal photosynthesis and possibly the polysaccharides of the biofilm matrix. Bacterial production of biofilm communities was significantly higher in light incubations (p <0.001). The greatest differences in production rates between light and dark incubations occurred between 8 and 24 h. Biomass-specific a- and b-glucosidase and b-xylosidase activities were stimulated by photosynthesis, with significantly greater activities occurring at hours 16 and 24 in the light treatment (p <0.01). The results indicate that algal photosynthesis can have a significant impact on bacterial productivity, biomass, biovolume, and enzyme production over longer time periods at low photon flux densities (?100 mmol m-2 s-1).     

Post‐Impoundment Biomass and Composition of Phytoplankton in the Yangtze River

Damming, and thus alteration of stream flow, promotes higher phytoplankton populations and encourages algal blooms (density >10⁶ cells L^–1) in the Three Gorges Reservoir (TGR). Phytoplankton composition and biomass were studied in the Yangtze River from March 2004 to May 2005. 107 taxa were identified. Diatoms were the dominant group, followed by Chlorophyta and Cyanobacteria. In the Yangtze River, algal abundance varied from 3.13 × 10³ to 3.83 × 10⁶ cells L^–1, and algal biomass was in the range of 0.06 to 659 mg C m^–3. Levels of nitrogen, phosphorus and silica did not show consistent longitudinal changes along the river and were not correlated with phytoplankton parameters. Phytoplankton abundance was negatively correlated with main channel discharge (Spearman r = –1.000, P < 0.01). Phytoplankton abundance and biomass in the Yangtze River are mainly determined by the hydrological conditions rather than by nutrient concentrations. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

In situ interactions between the effects of season,current velocity and pollution on a river biofilm

1. The development of periphytic algae and bacteria is controlled by a combination of interacting biotic processes and abiotic factors. Distinguishing between the selection pressure resulting from pollution and that of natural environmental factors is therefore one of the most critical aspects of assessing the impact of pollutants on the diversity and function of benthic microbial communities in natural ecosystems. 2. We studied how current velocity and season affect the ability of river biofilms to cope with complex chemical pollution. We compared the diversity, structure and production of periphytic algae and bacteria from four sampling zones with differing chemical water quality levels and different flow velocities over the course of two seasons (summer and winter). 3. The three factors tested all influenced biofilm development, but this depended on the biological variable being measured. Bacterial and algal densities were highly dependent on season and chemical water quality. Algal density was lower in summer than in winter, but bacterial density and production increased from upstream (reference) to downstream (polluted), and this increase was more marked in winter. The impact of chemical water quality was also dependent on the season. 4. An interaction between current velocity and pollution was also detected. During the summer, there was no difference in bacterial density or production between the upstream and downstream segments in the fast current zones, whereas both variables were higher downstream in the slow current zones. Such interactions between environmental factors and the impact of water quality on biofilms must be taken into account in assessments of the effects of chemicals on biofilm community structure and functioning in rivers.     

The effect of light direction and suspended cell concentrations on algal biofilm growth rates

Algae biofilms were grown in a semicontinuous flat plate biofilm photobioreactor to study the effects of light direction and suspended algal cell populations on algal biofilm growth. It was determined that, under the growth conditions and biofilm thicknesses studied, light direction had no effect on long-term algal biofilm growth (26 days); however, light direction did affect the concentration of suspended algal cells by influencing the photon flux density in the growth medium in the photobioreactors. This suspended algal cell population affected short-term (7 days) algae cell recruitment and algal biofilm growth, but additional studies showed that enhanced suspended algal cell populations did not affect biofilm growth rates over the long term (26 days). Studying profiles of light transmittance through biofilms as they grew showed that most of the light became attenuated by the biomass after just a few days of growth (88 % after 3 days). The estimated biofilm thicknesses after these few days of growth were approximately 150 μm. The light attenuation data suggests that, although the biofilms grew to 700–900 μm, under these light intensities, only the first few hundred micrometers of the biofilm is receiving enough light to be photosynthetically active. We postulate that this photosynthetically active layer of the biofilm grows adjacent to the light source, while the rest of the biofilm is in a stationary growth phase. The results of this study have implications for algal biofilm photobioreactor design and operation.     

Grazing pressure of herbivorous coral reef fishes on low coral-cover reefs

The impact of grazing by herbivorous fishes (Acanthuridae, Scaridae, and Pomacentridae) on low coral-cover reefs was assessed by measuring rates of benthic algal production and consumption on inshore and offshore reefs in the upper Florida Keys. Algal production rates, determined in situ with caged and uncaged experimental plates, were low (mean 1.05 g C m⁻² day⁻¹) and similar among reef types. Algal consumption rates were estimated using two different models, a detailed model incorporating fish bite rates and algal yield-per-bite for one species extrapolated to a guild-wide value, and a general regression relating fish biomass to algal consumption. Algal consumption differed among reef types: a majority of algal production was consumed on offshore reefs (55–100%), whereas consumption on inshore patch reefs was 31–51%. Spatial variation in algal consumption was driven by differences in herbivorous fish species composition, density, and size-structure among reef types. Algal consumption rates also varied temporally due to seasonal declines in bite rates and intermittent presence of large-bodied, vagile, schooling species. Spatial coherence of benthic community structure and temporal stability of algal turf over 3 years suggests that grazing intensity is currently sufficient to limit further spread of macroalgal cover on these low coral-cover reefs, but not to exclude it from the system.     

Diversity and Seasonal Variability of β-Proteobacteria in Biofilms of Polluted Rivers: Analysis by Temperature Gradient Gel Electrophoresis and Cloning

The β-subgroup of the Proteobacteria has been shown to be important in aquatic habitats and was investigated in depth here by molecular 16S rRNA techniques in biofilms of the Elbe River and its polluted tributary, the Spittelwasser River. The bacterial 16S rRNA genes were cloned from each site, screened for β-proteobacterial clones and sequenced. River biofilm clones from both rivers grouped into 9 clusters (RBFs). RBFs 1, 2, and 3 fell into the recently described βI cluster of cosmopolitan freshwater bacteria, where they represented new species related to Rhodoferax, Aquaspirillum, and Hydrogenophaga. RBFs 4 to 7 affiliated with Aquabacterium commune, Ideonella dechloratans, and Sphaerotilus natans, respectively. The two remaining RBFs were uncultivated clusters, one of them being distantly related to Gallionella ferruginea. Seasonal changes in the relative intensity of the β-proteobacterial 16S rRNA genes of biofilms harvested monthly for 1 year were determined by specific amplification and separation by temperature gradient gel electrophoresis (TGGE). Bands were identified by comparison of clones to community fingerprints by TGGE. Eight of 13 identified bands were shared by both habitats but showed different relative abundance and seasonal variability in the two rivers, probably caused by differences in temperature and pollutants. The data indicate new not-yet-cultivated clusters of river biofilm organisms, some of them probably distributed globally. They confirm the importance of certain known freshwater genera in river biofilms. The high phylogenetic resolution obtained by clone library analysis combined with the high temporal resolution obtained by TGGE suggest that the observed microdiversity in the river biofilm clone libraries might be caused by phylogenetically closely related microbial populations which are adapted to ecological parameters.