Distinct dynamics in mountain watersheds: Exploring mercury and microplastic pollution-Unraveling the influence of atmospheric deposition, human activities, and hydrology.

The intensification of human activities all around the globe has led to the spread of micropollutants in high-mountain freshwater environments. We therefore aimed to assess the geospatial distribution and determine the potential sources of (total-) mercury (THg) and microplastics (MPs) in mountain freshwater ecosystems. To do so, we analyzed THg and MP concentrations in brown trout, biofilm, and sediments from lotic and lentic ecosystems in the Pyrenees - all subjected to different types of human pressure. Additionally, we assessed the potential impacts of these pollutants on fish, and explored the bioindication capacity of brown trout (Salmo trutta fario) and biofilm regarding THg and MP pollution. For the first time, we measured concentrations of MPs trapped in the matrix of freshwater biofilm. Our results suggest that THg in the Pyrenees might be explained by both legacy (regional) and distant sources, in combination with environmental characteristics such as the presence of peatlands or streamwater physicochemistry, while MPs in fish are linked to recent local pollution sources such as single-use plastics. In contrast, MPs in biofilm matrix and sediments indicate a combination of distant (i.e., atmospheric deposition) and recent local pollution sources. Moreover, hydrodynamics and plastic density likely control MP distribution in rivers. Based on Fulton's condition factor, we also found that higher THg concentrations caused a negative impact on fish health (K < 1), while no impact of MPs could be seen. Therefore, we suggest that brown trout and biofilm can serve as bioindicators of atmospheric deposition of THg in high-altitude lakes and that biofilm is a reliable bioindicator to assess MP pollution in remote environments. Brown trout may also act as a bioindicator of MP pollution, but only efficiently in more polluted areas.

Benthic sediment as stores and sources of bacteria and viruses in streams: A comparison of baseflow vs. stormflow longitudinal transport and residence times.

The presence of bacteria and viruses in freshwater represents a global health risk. The substantial spatial and temporal variability of microbes leads to difficulties in quantifying the risks associated with their presence in freshwater. Fine particles, including bacteria and viruses are transported and accumulated into shallow streambed (i.e., benthic) sediment, delaying the downstream transmission during baseflow conditions but contributing to their resuspension and transport downstream during stormflow events. Direct measurements of pathogen accumulation in benthic sediments are rare. Until now, the dynamic role of benthic sediment as both a store and source of microbes, has not been quantified. In this study, we analyze microbial abundance in benthic sediment along a 1 km reach of an intermittent Mediterranean stream receiving inputs from the effluent of a wastewater treatment plant, a known point source of microbes in streams. We sampled benthic sediment during a summer drought when the wastewater effluent constituted 100 % of the stream flow, and thus, large accumulation and persistence of pathogens along the streambed was expected. We measured the abundance of total bacteria, Escherichia coli (as a fecal indicator), and presence of enteric rotavirus (RoV) and norovirus (NoV). The abundance of E. coli, based on qPCR detection, was high (4.99∙102 gc /cm2) along the first 100 m downstream of the wastewater effluent input and in general decreased with distance from the source, with presence of RoV and NoV along the study reach. A particle tracking model was applied, that uses stream water velocity as an input, and accounts for microbial exchange into, immobilization, degradation, and resuspension out of benthic sediment during baseflow and stormflow. Rates of exchange into benthic sediment were 3 orders of magnitude higher during stormflow, but residence times were proportionately lower, resulting in increased longitudinal connectivity from up to downstream during stormflow. Model simulations demonstrated mechanistically how the rates of exchange into and out of the benthic sediment resulted in benthic sediment to act as a store during baseflow and a source during stormflow.

Stream Hydrology Controls the Longitudinal Bioreactive Footprint of Urban-Sourced Fine Particles.

The relevance of wastewater treatment plant (WWTP) effluents in fluvial networks is increasing as urbanization grows in catchments. Urban-sourced fine particles from WWTP effluents deposit and accumulate in the streambed sediment of receiving streams over time and can fuel respiration rates, which can thus potentially increase rates of biogeochemical reactions and CO2 emissions to the atmosphere. We aimed to provide a quantitative assessment of the influence of WWTP-sourced fine particles deposited in the streambed sediment on stream metabolic activity for 1 year in an intermittent Mediterranean stream. More nutrient-rich and metabolically active fine particle standing stocks were observed downstream of the WWTP, propagating to the end of the 820 m study reach, especially during the dry period (i.e., when the dilution capacity of the stream to WWTP inputs is <40%). Based on the longitudinal patterns of fine particle standing stocks and their metabolic activity, we estimated that the in-stream bioreactive capacity associated with these fine particles could potentially lead to substantial carbon dioxide emissions to the atmosphere (3.1 g C/m2/d). We show the importance of incorporating fine particle standing stocks downstream of point source inputs, particularly WWTPs in intermittent streams, into carbon budgets.

Effect of Three Emergent Macrophyte Species on Nutrient Retention in Aquatic Environments under Excess Nutrient Loading.

Emergent macrophyte species selection is critical for the effectiveness of nature-based engineered solutions aiming to address excess nutrient concentrations in freshwater ecosystems. Yet, the mechanisms with which macrophytes enhance nutrient retention need to be further understood. Here, we compared nutrient retention among 12 artificial flumes fed with effluents from a wastewater treatment plant and subjected to four treatments: absence of macrophytes (control) and presence of three different macrophyte species (Iris pseudacorus L., Phragmites australis L., and Schoenoplectus lacustris L.). We estimated the net and gross nutrient uptake based on the longitudinal profiles of ambient concentrations and on pulse injections of ammonium (NH4+) and soluble reactive phosphorus. Further, we investigated the influence of subsurface hydrological retention, attributed to the architectural differences in the roots of these macrophytes, on nutrient retention. Results showed a species-specific effect of macrophytes on nutrient retention and confirmed root-associated subsurface hydrological retention as a driving factor. Schoenoplectus showed both high net and gross NH4+ uptake, thereby being the most effective species to address N loading, compared to Iris and Phragmites. This work contributes to improve our mechanistic understanding of the role of emergent macrophytes on nutrient retention in aquatic environments.

Wastewater treatment plant effluent inputs induce large biogeochemical changes during low flows in an intermittent stream but small changes in day-night patterns.

Wastewater treatment plant (WWTP) effluents alter stream water chemistry and metabolic activity. Yet, essential aspects influencing the biogeochemical response of receiving streams such as hydrology and diel oscillations of light and temperature remain largely unexplored. We measured day vs night water chemistry and in-stream net nutrient uptake velocity (Vf) in an intermittent forested stream, upstream and downstream of a WWTP effluent under contrasting hydrological conditions. The WWTP effluent negatively influenced stream water chemistry, especially during the dry period. Despite large diel oscillations in light inputs, day-night differences in nutrient and oxygen concentrations were small, suggesting that heterotrophic respiration drove stream metabolism with a minor contribution of gross primary production. The magnitude of Vf was similar between day and night at the two reaches. Yet, at the downstream reach, in-stream net DIN uptake occurred more often at night, and values of Vf for ammonia and nitrite indicated enhanced in-stream nitrification. The two reaches showed a small capacity to retain DIN and soluble reactive phosphorus from the water column. Positive values of in-stream net nutrient uptake (i.e. uptake > release) occurred mostly during the dry period, highlighting that in-stream biogeochemical processing can contribute to improve water quality in streams receiving point-sources effluents in regions with low water availability.

The method controls the story - Sampling method impacts on the detection of pore-water nitrogen concentrations in streambeds.

Biogeochemical gradients in streambeds are steep and can vary over short distances often making adequate characterisation of sediment biogeochemical processes challenging. This paper provides an overview and comparison of streambed pore-water sampling methods, highlighting their capacity to address gaps in our understanding of streambed biogeochemical processes. This work reviews and critiques available pore-water sampling techniques to characterise streambed biogeochemical conditions, including their characteristic spatial and temporal resolutions, and associated advantages and limitations. A field study comparing three commonly-used pore-water sampling techniques (multilevel mini-piezometers, miniature drivepoint samplers and diffusive equilibrium in thin-film gels) was conducted to assess differences in observed nitrate and ammonium concentration profiles. Pore-water nitrate concentrations did not differ significantly between sampling methods (p-value = 0.54) with mean concentrations of 2.53, 4.08 and 4.02 mg l-1 observed with the multilevel mini-piezometers, miniature drivepoint samplers and diffusive equilibrium in thin-film gel samplers, respectively. Pore-water ammonium concentrations, however, were significantly higher in pore-water extracted by multilevel mini-piezometers (3.83 mg l-1) and significantly lower where sampled with miniature drivepoint samplers (1.05 mg l-1, p-values <0.01). Differences in observed pore-water ammonium concentration profiles between active (suction: multilevel mini-piezometers) and passive (equilibrium; diffusive equilibrium in thin-film gels) samplers were further explored under laboratory conditions. Measured pore-water ammonium concentrations were significantly greater when sampled by diffusive equilibrium in thin-film gels than with multilevel mini-piezometers (all p-values ≤0.02). The findings of this study have critical implications for the interpretation of field-based research on hyporheic zone biogeochemical cycling and highlight the need for more systematic testing of sampling protocols. For the first time, the impact of different active and passive pore-water sampling methods is addressed systematically here, highlighting to what degree the choice of pore-water sampling methods affects research outcomes, with relevance for the interpretation of previously published work as well as future studies.

The role of helophyte species on nitrogen and phosphorus retention from wastewater treatment plant effluents.

In the Mediterranean region, water scarcity compromises stream water quality particularly downstream of wastewater treatment plants (WWTP). We tested the potential of four helophyte species to reduce dissolved inorganic nitrogen (N) and phosphorus (P) from WWTP effluents. We conducted an 11-month mesocosm experiment to assess differences in N and P content among plant compartments and among species. Moreover, we quantified the relative contribution of above and belowground parts of the plants to N and P retention. The experiment was conducted at the Urban River Laboratory (www.urbanriverlab.com) in artificial channels (12 m long x 0.6 m wide x 0.4 m deep) planted with monospecific stands of Iris pseudoacorus, Typha angustifolia, Phragmites australis and Scirpus lacustris. Channels (three replicates per species) received water from the WWTP effluent, which flowed at a constant rate of 5 L min-1 through the sub-surface. The helophytes were planted in November 2014 and biomass standing stocks of carbon (C), N and P were measured in October 2015 at the time of maximum plant biomass. Differences in the concentration of N and P were larger among plant compartments than among species. The highest N concentration was measured in leaves while rhizomes showed the highest P concentration. The total plant biomass varied greatly among species from 11.4 to 4.6 Kg DW m-2 for Iris and Scirpus, respectively. Iris accumulated the highest amount of N (256 g N m-2) and P (27 g P m-2) in biomass. Plants retained from 8% (Scirpus) to 19% (Iris) of total dissolved inorganic N inputs to the channels (10.4 kg N) during the experiment, and from 6% (Phragmites) to 14% (Iris) of total dissolved inorganic P inputs (1.3 kg P). This study provides quantitative evidence to water managers of the potential role of helophytes to improve water quality in freshwater ecosystems receiving water from WWTP effluents.

Leachates from Helophyte Leaf-Litter Enhance Nitrogen Removal from Wastewater Treatment Plant Effluents.

Bioengineering techniques are currently used  in a wide variety of wastewater treatment systems. Aquatic plants (i.e., helophytes) used in these techniques reduce excess nitrogen (N) from water column via assimilation. Moreover, leachates from plant leaf-litter can serve as an additional source of labile dissolved organic matter (DOM), which can promote aerobic respiration and N removal via denitrification. We tested the influence of leaf-litter leachates from  Iris pseudacorus and Phragmites australis on the structure and activity of freshwater biofilms grown in flumes fed by effluent from a wastewater treatment plant (WWTP). The responses of the epilithic biofilm to the inputs of leaf-litter leachates were compared to those measured using a brewery byproduct rich in sugars and to the WWTP effluent water (i.e., control). All DOM sources significantly enhanced aerobic respiration and denitrification of the biofilm when compared to the controls, with increases in total microbial abundance but not in denitrifier abundance. The results suggest that metabolic activity of biofilms may be limited by bioavailability of DOM in WWTP effluent; and leaf-litter leachates of helophytes used in bioengineering techniques could alleviate this limitation by enhancing microbial N and C uptake.

Drivers of nitrogen transfer in stream food webs across continents.

Studies of trophic-level material and energy transfers are central to ecology. The use of isotopic tracers has now made it possible to measure trophic transfer efficiencies of important nutrients and to better understand how these materials move through food webs. We analyzed data from thirteen 15 N-ammonium tracer addition experiments to quantify N transfer from basal resources to animals in headwater streams with varying physical, chemical, and biological features. N transfer efficiencies from primary uptake compartments (PUCs; heterotrophic microorganisms and primary producers) to primary consumers was lower (mean 11.5%, range <1% to 43%) than N transfer efficiencies from primary consumers to predators (mean 80%, range 5% to >100%). Total N transferred (as a rate) was greater in streams with open compared to closed canopies and overall N transfer efficiency generally followed a similar pattern, although was not statistically significant. We used principal component analysis to condense a suite of site characteristics into two environmental components. Total N uptake rates among trophic levels were best predicted by the component that was correlated with latitude, DIN:SRP, GPP:ER, and percent canopy cover. N transfer efficiency did not respond consistently to environmental variables. Our results suggest that canopy cover influences N movement through stream food webs because light availability and primary production facilitate N transfer to higher trophic levels.

Combined effects of hydrologic alteration and cyprinid fish in mediating biogeochemical processes in a Mediterranean stream.

Flow regimes are important drivers of both stream community and biogeochemical processes. However, the interplay between community and biogeochemical responses under different flow regimes in streams is less understood. In this study, we investigated the structural and functional responses of periphyton and macroinvertebrates to different densities of the Mediterranean barbel (Barbus meridionalis, Cyprinidae) in two stream reaches differing in flow regime. The study was conducted in Llémena Stream, a small calcareous Mediterranean stream with high nutrient levels. We selected a reach with permanent flow (permanent reach) and another subjected to flow regulation (regulated reach) with periods of flow intermittency. At each reach, we used in situ cages to generate 3 levels of fish density. Cages with 10 barbels were used to simulate high fish density (>7indm-2); cages with open sides were used as controls (i.e. exposed to actual fish densities of each stream reach) thus having low fish density; and those with no fish were used to simulate the disappearance of fish that occurs with stream drying. Differences in fish density did not cause significant changes in periphyton biomass and macroinvertebrate density. However, phosphate uptake by periphyton was enhanced in treatments lacking fish in the regulated reach with intermittent flow but not in the permanent reach, suggesting that hydrologic alteration hampers the ability of biotic communities to compensate for the absence of fish. This study indicates that fish density can mediate the effects of anthropogenic alterations such as flow intermittence derived from hydrologic regulation on stream benthic communities and associated biogeochemical processes, at least in eutrophic streams.

Enhancement of carbon and nitrogen removal by helophytes along subsurface water flowpaths receiving treated wastewater.

Wastewater treatment plant (WWTP) effluents are sources of dissolved organic carbon (DOC) and inorganic nitrogen (DIN) to receiving streams, which can eventually become saturated by excess of DIN. Aquatic plants (i.e., helophytes) can modify subsurface water flowpaths as well as assimilate nutrients and enhance microbial activity in the rhizosphere, yet their ability to increase DIN transformation and removal in WWTP-influenced streams is poorly understood. We examined the influence of helophytes on DIN removal along subsurface water flowpaths and how this was associated with DOC removal and labile C availability. To do so, we used a set of 12 flow-through flumes fed with water from a WWTP effluent. The flumes contained solely sediments or sediments with helophytes. Presence of helophytes in the flumes enhanced both DIN and DOC removal. Experimental addition of a labile C source into the flumes resulted in a high removal of the added C within the first meter of the flumes. Yet, no concomitant increases in DIN removal were observed. Moreover, results from laboratory assays showed significant increases in the potential denitrifying enzyme activity of sediment biofilms from the flumes when labile C was added; suggesting denitrification was limited by C quality. Together these results suggest that lack of DIN removal response to the labile C addition in flumes was likely because potential increases in denitrification by biofilms from sediments were counterbalanced by high rates of mineralization of dissolved organic matter. Our results highlight that helophytes can enhance DIN removal in streams receiving inputs from WWTP effluents; and thus, they can become a relevant bioremediation tool in WWTP-influenced streams. However, results also suggest that the quality of DOC from the WWTP effluent can influence the N removal capacity of these systems.

Stream drying drives microbial ammonia oxidation and first-flush nitrate export.

Stream microbial communities and associated processes are influenced by environmental fluctuations that may ultimately dictate nutrient export. Discharge fluctuations caused by intermittent stream flow are increasing worldwide in response to global change. We examined the impact of flow cessation and drying on in-stream nitrogen cycling. We determined archaeal (AOA) and bacterial ammonia oxidizer (AOB) abundance and ammonia oxidation activity in surface and deep sediments from different sites along the Fuirosos stream (Spain) subjected to contrasting hydrological conditions (i.e., running water, isolated pools, and dry streambeds). AOA were more abundant than AOB, with no major changes across hydrological conditions or sediment layers. However, ammonia oxidation activity and sediment nitrate content increased with the degree of stream drying, especially in surface sediments. Upscaling of our results shows that ammonia oxidation in dry streambeds can contribute considerably (~50%) to the high nitrate export typically observed in intermittent streams during first-flush events following flow reconnection. Our study illustrates how the dry channels of intermittent streams can be potential hotspots of ammonia oxidation. Consequently, shifts in the duration, spatial extent and severity of intermittent flow can play a decisive role in shaping nitrogen cycling and export along fluvial networks in response to global change.

Green light: gross primary production influences seasonal stream N export by controlling fine-scale N dynamics.

Monitoring nutrient concentrations at fine-scale temporal resolution contributes to a better understanding of nutrient cycling in stream ecosystems. However, the mechanisms underlying fine-scale nutrient dynamics and its implications for budget catchent fluxes are still poorly understood. To gain understanding of patterns and controls of fine-scale stream nitrogen (N) dynamics and to assess how they affect hydrological N fluxes, we explored diel variation in stream nitrate (NO3-) concentration along a headwater stream with increasing riparian area and channel width. At the downstream site, the highest day-night variations occurred in early spring, when stream NO3- concentrations were 13% higher at night than at daytime. Such day-night variations were strongly related to daily light inputs (R2 = 0.74) and gross primary production (GPP; R2 = 0.74), and they showed an excellent fit with day-night NO- variations predicted from GPP (R2 = 0.85). These results suggest that diel fluctuations in stream NO3- concentration were mainly driven by photoautotrophic N uptake. Terrestrial influences were discarded because no simultaneous diel variations in stream discharge, riparian groundwater level, or riparian solute concentration were observed. In contrast to the downstream site, no diel variations in NO3- concentration occurred at the upstream site, likely because water temperature was colder (10 degrees C vs. 12 degrees C) and light availability was lower (4 vs. 9 mol x m(-2) x d(-1)). Although daily GPP was between 10- and 100-fold lower than daily respiration, photoautotrophic N uptake contributed to a 10% reduction in spring NO3- loads at the downstream site. Our study clearly shows that the activity of photoautotrophs can substantially change over time and along the stream continuum in response to key environmental drivers such as light and temperature, and further, that its capacity to regulate diel and seasonal N fluxes can be important even in low-productivity streams.

Resource subsidies between stream and terrestrial ecosystems under global change.

Streams and adjacent terrestrial ecosystems are characterized by permeable boundaries that are crossed by resource subsidies. Although the importance of these subsidies for riverine ecosystems is increasingly recognized, little is known about how they may be influenced by global environmental change. Drawing from available evidence, in this review we propose a conceptual framework to evaluate the effects of global change on the quality and spatiotemporal dynamics of stream-terrestrial subsidies. We illustrate how changes to hydrological and temperature regimes, atmospheric CO2 concentration, land use and the distribution of nonindigenous species can influence subsidy fluxes by affecting the biology and ecology of donor and recipient systems and the physical characteristics of stream-riparian boundaries. Climate-driven changes in the physiology and phenology of organisms with complex life cycles will influence their development time, body size and emergence patterns, with consequences for adjacent terrestrial consumers. Also, novel species interactions can modify subsidy dynamics via complex bottom-up and top-down effects. Given the seasonality and pulsed nature of subsidies, alterations of the temporal and spatial synchrony of resource availability to consumers across ecosystems are likely to result in ecological mismatches that can scale up from individual responses, to communities, to ecosystems. Similarly, altered hydrology, temperature, CO2 concentration and land use will modify the recruitment and quality of riparian vegetation, the timing of leaf abscission and the establishment of invasive riparian species. Along with morphological changes to stream-terrestrial boundaries, these will alter the use and fluxes of allochthonous subsidies associated with stream ecosystems. Future research should aim to understand how subsidy dynamics will be affected by key drivers of global change, including agricultural intensification, increasing water use and biotic homogenization. Our conceptual framework based on the match-mismatch between donor and recipient organisms may facilitate understanding of the multiple effects of global change and aid in the development of future research questions.

Satisfaction and quality of life with palatal positioned implants in severely atrophic maxillae versus conventional implants supporting fixed full-arch prostheses.

BACKGROUND: To evaluate satisfaction and quality of life in patients with palatal positioned implants supporting fixed full-arch prostheses to rehabilitate edentulous maxillae with horizontal atrophy and compare them with conventional well-centered implants placed in non-atrophic supporting fixed full-arch prostheses. MATERIAL AND METHOD: A clinical retrospective study was performed of patients that were rehabilitated with full-arch fixed implant-supported maxillary prostheses and had a minimum follow-up of 5 years after implant loading. Patients were divided into 2 groups: patients with class IV maxilla according to Cawood and Howell and treated with palatal positioned implants (test) and with class III maxilla and treated with implants well-centered in the alveolar ridge and completely surrounded by bone (control). Ten-cm visual analogue scales (VAS) (range 1-10) and the OHIP-14 (Oral Health Impact Profile) questionnaire were used respectively to estimate patient satisfaction and quality of life after implant therapy. Statistical analysis was performed applying Mann-Whitney Test using alpha set at 0.05. RESULTS: Mean global and specific satisfaction--except for self-esteem--were superior for the test group than the control group, although differences were not statistically significant. Regarding quality of life, the reported incidence of problems was lower in the test group for all the studied items except for 'problems at work'. However, differences were not statistically significant in any case. CONCLUSION: Despite the limitations of the study (retrospective and nonrandomized design) the results suggest that the prosthesis design needed to rehabilitate palatally positioned implants (more coverage of palate) does not lead to lower satisfaction and quality of life of patients, compared to patients treated with implants placed centered and conventional design prostheses that do not cover the palate.

Palatal positioned implants in severely atrophic maxillae versus conventional implants to support fixed full-arch prostheses: Controlled retrospective study with 5 years of follow-up.

BACKGROUND: To evaluate soft tissue conditions and bone loss around palatal positioned implants supporting fixed full-arch prostheses to rehabilitate edentulous maxillae with horizontal atrophy and compare them with conventional well-centered implants placed in non-atrophic maxillae after a minimum follow-up of 5 years. MATERIAL AND METHODS: A clinical retrospective study was performed of patients that were rehabilitated with full-arch fixed implant-supported maxillary prostheses and had a minimum follow-up of 5 years after implant loading. Patients were divided into 2 groups: patients with class IV maxilla according to Cawood and Howell and treated with palatal positioned implants (test) and with class III maxilla and treated with implants well-centered in the alveolar ridge and completely surrounded by bone (control). The following variables were assessd: age, sex, frequency of toothbrushing, smoking, type of prosthesis, type of implant, implant success, amount of buccal keratinized mucosa, buccal retraction, probing depth, plaque index, modified bleeding index, presence of mucositis or peri-implantitis and peri-implant bone loss. Statistical analysis was performed applying Chi2 Test and Student's t-test using alpha set at 0.05. RESULTS: A total of 57 patients were included: 32 patients with 161 palatal positioned implants (test) and 25 patients with 132 well centered implants (control). No statistically significant differences were found regarding age, sex and smoking, but test group patients reported a significantly higher frequency of daily toothbrushing. Implant success rates were 96.9% for test group implants and 96.0% for control group implants. Peri-implant mucosa retraction was significantly higher in the control group than in the test group (p=0,017). No significant differences were observed either for all the other assessed clinical parameters or for peri-implant bone loss. CONCLUSIONS: Despite its limitations the outcomes of the present study suggest that palatal positioned implants may be a good treatment alternative for patients with severe horizontal maxillary alveolar bone atrophy. Palatal positioned implants presented similar success rates, soft tissue conditions and peri-implant bone loss than well-centered implants placed completely surrounded by bone in non-atrophic ridges.

Wastewater treatment plant effluents change abundance and composition of ammonia-oxidizing microorganisms in mediterranean urban stream biofilms.

Streams affected by wastewater treatment plant (WWTP) effluents are hotspots of nitrification. We analyzed the influence of WWTP inputs on the abundance, distribution, and composition of epilithic ammonia-oxidizing (AO) assemblages in five Mediterranean urban streams by qPCR and amoA gene cloning and sequencing of both archaea (AOA) and bacteria (AOB). The effluents significantly modified stream chemical parameters, and changes in longitudinal profiles of both NH(4)(+) and NO(3)(-) indicated stimulated nitrification activity. WWTP effluents were an allocthonous source of both AOA, essentially from the Nitrosotalea cluster, and mostly of AOB, mainly Nitrosomonas oligotropha, Nitrosomonas communis, and Nitrosospira spp. changing the relative abundance and the natural composition of AO assemblages. Under natural conditions, Nitrososphaera and Nitrosopumilus AOA dominated AO assemblages, and AOB were barely detected. After the WWTP perturbation, epilithic AOB increased by orders of magnitude whereas AOA did not show quantitative changes but a shift in population composition to dominance of Nitrosotalea spp. The foraneous AOB successfully settled in downstream biofilms and probably carried out most of the nitrification activity. Nitrosotalea were only observed downstream and only in biofilms exposed to either darkness or low irradiance. In addition to other potential environmental limitations for AOA distribution, this result suggests in situ photosensitivity as previously reported for Nitrosotalea under laboratory conditions.

Stream carbon and nitrogen supplements during leaf litter decomposition: contrasting patterns for two foundation species.

Leaf litter decomposition plays a major role in nutrient dynamics in forested streams. The chemical composition of litter affects its processing by microorganisms, which obtain nutrients from litter and from the water column. The balance of these fluxes is not well known, because they occur simultaneously and thus are difficult to quantify separately. Here, we examined C and N flow from streamwater and leaf litter to microbial biofilms during decomposition. We used isotopically enriched leaves ((13)C and (15)N) from two riparian foundation tree species: fast-decomposing Populus fremontii and slow-decomposing Populus angustifolia, which differed in their concentration of recalcitrant compounds. We adapted the isotope pool dilution method to estimate gross elemental fluxes into litter microbes. Three key findings emerged: litter type strongly affected biomass and stoichiometry of microbial assemblages growing on litter; the proportion of C and N in microorganisms derived from the streamwater, as opposed to the litter, did not differ between litter types, but increased throughout decomposition; gross immobilization of N from the streamwater was higher for P. fremontii compared to P. angustifolia, probably as a consequence of the higher microbial biomass on P. fremontii. In contrast, gross immobilization of C from the streamwater was higher for P. angustifolia, suggesting that dissolved organic C in streamwater was used as an additional energy source by microbial assemblages growing on slow-decomposing litter. These results indicate that biofilms on decomposing litter have specific element requirements driven by litter characteristics, which might have implications for whole-stream nutrient retention.

Temporal variability of nitrogen stable isotopes in primary uptake compartments in four streams differing in human impacts.

Understanding the variability of the natural abundance in nitrogen stable isotopes (expressed as δ(15)N) of primary uptake compartments (PUCs; e.g., epilithon or macrophytes) is important due to the multiple applications of stable isotopes in freshwater research and can give insights into environmental and anthropogenic factors controlling N dynamics in streams. While previous research has shown how δ(15)N of PUCs varies with δ(15)N of dissolved inorganic N (DIN) among streams, less is known about how δ(15)N of PUCs varies over time. Here, we examined monthly variation of δ(15)N of PUCs and of DIN species (nitrate and ammonium) over a year, and compared it among streams with contrasting human impacts and PUC types. Our results showed no evidence of isotopic seasonal patterns. Temporal variability in δ(15)N-PUCs increased with human impact, being the highest in the urban stream, probably influenced by the high variability of δ(15)N-DIN. Among compartments, in-stream PUCs characterized by fast turnover rates, such as filamentous algae, showed the highest temporal variability in δ(15)N values (from -3.6 to 23.2 ‰). Our study elucidates some of the environmental and biological controls of temporal variability of δ(15)N in streams, which should be taken into account when using stable isotopes as an ecological tool.