Study Of Aquatic Systems Research Articles

Geology Can Drive the Diversity-Ecosystem Functioning Relationship in River Benthic Diatoms by Selecting for Species Functional Traits.

The biodiversity-ecosystem functioning (BEF) relationship has been studied extensively for the past 30 years, mainly in terrestrial plant ecosystems using experimental approaches. Field studies in aquatic systems are scarce, and considering primary producers, they mainly focus on phytoplankton assemblages, whereas benthic diatoms in rivers are considerably understudied in this regard. We performed a field study across nine rivers in Greece, and we coupled the observed field results with model simulations. We tested the hypothesis that the diversity-biomass (as a surrogate of ecosystem functioning) relationship in benthic diatoms would be affected by abiotic factors and would be time-dependent due to the highly dynamic nature of rivers. Indeed, geology played an important role in the form of the BEF relationship that was positive in siliceous and absent in calcareous substrates. Geology was responsible for nutrient concentrations, which, in turn, were responsible for the dominance of specific functional traits. Furthermore, model simulations showed the time dependence of the BEF form, as less mature assemblages tend to present a positive BEF. This was the first large-scale field study on the BEF relationship of benthic diatom assemblages, offering useful insights into the function and diversity of these overlooked ecosystems and assemblages.

Multi-Biomarker Analysis for Identifying Organic Matter Sources in Small Mountainous River Watersheds: A Case Study of the Yuba River Watershed

Organic matter in soils and sediments derives from a mixture of biological origins, often making it difficult to determine inputs from individual sources. Complicating the determination of source inputs to soil and sedimentary organic matter (OM) is the fact that physical and microbial processes have likely modified the initial composition of these sources. This study focused on identifying the composition of watershed-derived OM to better understand inputs to inland waters and improve our ability to resolve between terrigenous and aquatic sources in downstream systems, such as estuaries and coasts. We surveyed OM sources from the Yuba River watershed in northern California to identify specific biomarkers that represent aquatic and terrigenous OM sources. Multiple classes of organic proxies—including sterols, fatty acids (FA), lignin phenols and stable carbon and nitrogen isotope values (δ13C, δ15N)—were measured in soils, vegetation, charcoal, and freshwater plankton to characterize representative source endmembers. Sterols—including 27-nor-24-cholesta-5,22-dien-3β-ol, cholesta-5,22-dien-3β-ol, 24-methylcholesta-5,22-dien-3β-ol and cholesta-5-en-3β-ol, and positive δ15N values—were associated with aquatic OM (plankton, suspended particulate OM), whereas lignin phenols, long chain FA, and diacids characterized terrigenous sources (soils, charcoal, vegetation). Trends in organic carbon and biomarker signatures in soil samples showed a response to environmental disturbance (i.e., mining, agriculture) through an inverse relationship between OM content and land use. Results from this study demonstrate the utility of multi-biomarker studies for distinguishing between OM from different sources and land uses, offering new insights for biogeochemical studies in aquatic systems.

Multi-Biomarker Analysis for Identifying Organic Matter Sources in Small Mountainous River Watersheds: A Case Study of the Yuba River Watershed

Organic matter in soils and sediments derives from a mixture of biological origins, often making it difficult to determine inputs from individual sources. Complicating the determination of source inputs to soil and sedimentary organic matter (OM) is the fact that physical and microbial processes have likely modified the initial composition of these sources. This study focused on identifying the composition of watershed-derived OM to better understand inputs to inland waters and improve our ability to resolve between terrigenous and aquatic sources in downstream systems, such as estuaries and coasts. We surveyed OM sources from the Yuba River watershed in northern California to identify specific biomarkers that represent aquatic and terrigenous OM sources. Multiple classes of organic proxies—including sterols, fatty acids (FA), lignin phenols and stable carbon and nitrogen isotope values (δ13C, δ15N)—were measured in soils, vegetation, charcoal, and freshwater plankton to characterize representative source endmembers. Sterols—including 27-nor-24-cholesta-5,22-dien-3β-ol, cholesta-5,22-dien-3β-ol, 24-methylcholesta-5,22-dien-3β-ol and cholesta-5-en-3β-ol, and positive δ15N values—were associated with aquatic OM (plankton, suspended particulate OM), whereas lignin phenols, long chain FA, and diacids characterized terrigenous sources (soils, charcoal, vegetation). Trends in organic carbon and biomarker signatures in soil samples showed a response to environmental disturbance (i.e., mining, agriculture) through an inverse relationship between OM content and land use. Results from this study demonstrate the utility of multi-biomarker studies for distinguishing between OM from different sources and land uses, offering new insights for biogeochemical studies in aquatic systems.

Machine learning applied to lentic habitat use by spawning walleye demonstrates the benefits of considering multiple spatial scales in aquatic research

Scale and hierarchy have received less attention in aquatic compared to terrestrial systems. Walleye (Sander vitreus) spawning habitat offers an opportunity to investigate scale’s importance. We estimated lake-, transect-, and quadrat-scale influences on nearshore walleye egg deposition in 28 Minnesota lakes from 2016–2018. Random forest models (RFM) estimated importance of predictive variables to walleye egg deposition. Predictive accuracies of a multi-scale classification tree (CT) and a quadrat-scale CT were compared. RFM results suggested that five of our variables were unimportant when predicting egg deposition. The multi-scale CT was more accurate than the quadrat-scale CT when predicting egg deposition. Both model results suggest that in-lake egg deposition by walleye is regulated by hierarchical abiotic processes and that silt–clay abundance at the transect-scale (reef-scale) is more important than abundance at the quadrat-scale (within-reef). Our results show machine learning can be used for scale-optimization and potentially to determine cross-scale interactions. Further incorporation of scale and hierarchy into studies of aquatic systems will increase our understanding of species–habitat relationships, especially in lentic systems where multi-scale approaches are rarely used.

The Joint Contributions of Environmental Filtering and Spatial Processes to Macroinvertebrate Metacommunity Dynamics in the Alpine Stream Environment of Baima Snow Mountain, Southwest China

While macroinvertebrates are extensively investigated in many river ecosystems, meta-community ecology perspectives in alpine streams are very limited. We assessed the role of ecological factors and temporal dynamics in the macroinvertebrate meta-community assembly of an alpine stream situated in a dry-hot valley of Baima Snow Mountain, China. We found that spatial structuring and environmental filtering jointly drive the structure of macroinvertebrate meta-community, with relative contributions to the variance in community composition changing over time. RDA ordination and variation partitioning indicate that environmental variables are the most important predictors of community organization in most scenarios, whereas spatial determinants also play a significant role. Moreover, the explanatory power, identity, and the relative significance of ecological factors change over time. Particularly, in the years 2018 and 2019, stronger environmental filtering was found shaping community assembly, suggesting that deterministic mechanisms predominated in driving community dynamics. However, spatial factors had a stronger predictive power on meta-community structures in 2017, implying conspicuous dispersal mechanisms which may be owing to increased connectivity amongst sites. Thereby, we inferred that the alpine stream macroinvertebrate metacommunity composition can be regulated by the interaction of both spatial processes and environmental filtering, with relative contributions varying over time. Based on these findings, we suggest that community ecology studies in aquatic systems should be designed beyond single snapshot investigations.

Density‐Dependent Changes in Grass Carp Growth and Mortality in Long‐Term Aquatic Plant Management

AbstractThe Grass Carp Ctenopharyngodon idella is an herbivorous fish that has been stocked throughout the United States for aquatic plant control, commonly to manage hydrilla Hydrilla verticillata. Triploid Grass Carp were purported to have a mean life span of about 11 years based on otolith‐assigned ages and mortality estimates for fish associated with large stocking efforts through which aquatic plants were eliminated. Recent studies in aquatic systems where preferred foods were not eliminated have found that Grass Carp may live up to 30 years or more, but information about growth and mortality is lacking. About 130,000 Grass Carp have been stocked since 1995 to manage hydrilla in combination with fluridone application in Lake Gaston, Virginia–North Carolina, USA. Hydrilla coverage peaked at about 1,300 ha in 2003 and was reduced to about 130 ha by 2014. Since 2006, a total of 419 Grass Carp have been collected for age and growth analysis. A Bayesian approach was used to estimate von Bertalanffy growth function parameters that incorporated the effects of a linear trend in year, hydrilla abundance, or Grass Carp density and to assess population changes. Fish that were stocked in 1995 persisted through 2017. Mean maximum length (L∞) of Grass Carp decreased by about 200 mm with an increasing number of Grass Carp per hectare of hydrilla, and fish reached maximum size earlier in life. Indirect mortality estimates derived from growth parameters increased with Grass Carp density. Because preferred food availability affects mortality rates, stocking numbers should vary accordingly to maximize the effectiveness of hydrilla control programs.

Optimizing an eDNA protocol for estuarine environments: Balancing sensitivity, cost and time

Environmental DNA (eDNA) analysis has gained traction as a precise and cost-effective method for species and waterways management. To date, publications on eDNA protocol optimization have focused primarily on DNA yield. Therefore, it has not been possible to evaluate the cost and speed of specific components of the eDNA protocol, such as water filtration and DNA extraction method when designing or choosing an eDNA protocol. At the same time, these two parameters are essential for the experimental design of a project. Here we evaluate and rank 27 different eDNA protocols in the context of Chinook salmon (Oncorhynchus tshawytscha) eDNA detection in an estuarine environment. We present a comprehensive evaluation of multiple eDNA protocol parameters, balancing time, cost and DNA yield. We collected samples composed of 500 mL estuarine water from Deverton Slough (38°11'16.7"N 121°58'34.5"W) and 500 mL from tank water containing 1.3 juvenile Chinook Salmon per liter. Then, we compared extraction methods, filter types, use of inhibitor removal kit for DNA yield, processing time, and protocol cost. Lastly, we used an MCMC algorithm together with machine learning to understand the DNA yield of each step of the protocol as well as the interactions between those steps. Glass fiber filtration was to be the most resilient to high turbidites, filtering the samples in 2.32 ± 0.08 min instead of 14.16 ± 1.86 min and 6.72 ± 1.99 min for nitrocellulose and paper filter N1, respectively. The filtration DNA yield percentages for paper filter N1, glass fiber, and nitrocellulose were 0.00045 ± 0.00013, 0.00107 ± 0.00013, 0.00172 ± 0.00013. The DNA extraction yield percentage for QIagen, dipstick, NaOH, magnetic beads, and direct dipstick ranged from 0.047 ± 0.0388 to 0.475 ± 0.0357. For estuarine waters, which are challenging for eDNA studies due to high turbidity, variable salinity, and the presence of PCR inhibitors, we found that a protocol combining glass filters, magnetic beads, and an extra step for PCR inhibitor removal, is the method that best balances time, cost, and yield. In addition, we provide a generalized decision tree for determining the optimal eDNA protocol for other studies in aquatic systems. Our findings should be applicable to most aquatic environments and provide a clear guide for determining which eDNA protocol should be used under different study constraints.

Listening to air–water gas exchange in running waters

AbstractAir–water gas exchange velocities (k) are critical components of many biogeochemical and ecological process studies in aquatic systems. However, their high spatiotemporal variability is difficult to capture with traditional methods, especially in turbulent flow. Here, we investigate the potential of sound spectral analysis to infer k in running waters, based on the rationale that both turbulence and entrained bubbles drive gas exchange and cause a characteristic sound. We explored the relationship between k and sound spectral properties using laboratory experiments and field observations under a wide range of turbulence and bubble conditions. We estimated k using flux chamber measurements of CO2 exchange and recorded sound above and below the water surface by microphones and hydrophones, respectively. We found a strong influence of turbulence and bubbles on sound pressure levels (SPLs) at octave bands of 31.5 Hz and 1000 Hz, respectively. The difference in SPLs at these bands and background noise bands showed a linear correlation with k both in the laboratory (R2 = 0.93–0.99) and in the field (median R2 = 0.42–0.90). Underwater sound indices outperformed aerial sound indices in general, and indices based on hydraulic parameters in particular, in turbulent and bubbly surface flow. The results highlight the unique potential of acoustic techniques to predict k, isolate mechanisms, and improve the spatiotemporal coverage of k estimates in bubbly flow.

The case for reporting PO2 (partial pressure of oxygen), in addition to DO (dissolved oxygen), in studies of aquatic systems

In this essay, we question the sole use of mg/L (ppm) and the term DO (dissolved oxygen) when referring to oxygen supply in water, and discuss reasons for also reporting the partial pressure of oxygen (PO2). Oxygen moves from water into organisms by diffusion only, and the rate and direction of diffusion is solely dependent upon partial pressure gradients, not concentration gradients. However, the amount of oxygen moved into an organism is both a function of DO and PO2. Therefore, to have physiological and ecological relevance, the presence of oxygen in water, when reported as DO, should also include PO2.

Microplastic Abundance and Composition in Western Lake Superior As Determined via Microscopy, Pyr-GC/MS, and FTIR.

While plastic pollution in marine and freshwater systems is an active area of research, there is not yet an in-depth understanding of the distributions, chemical compositions, and fates of plastics in aquatic environments. In this study, the magnitude, distribution, and common polymers of microplastic pollution in surface waters in western Lake Superior are determined. Analytical methodology, including estimates of ambient contamination during sample collection and processing,are described and employed. Microscopy, pyrolysis-gas chromatography/mass spectrometry (Pyr-GC/MS), and Fourier transform infrared spectroscopy (FTIR) were used to quantify and identify microplastic particles. In surface waters, fibers were the most frequently observed morphology, and, based upon PyGC/MS analysis, polyvinyl chloride was the most frequently observed polymer, followed by polypropylene and polyethylene. The most common polymer identified by FTIR was polyethylene. Despite the low human population in Lake Superior's watershed, microplastic particles (particularly fibers, fragments, and films) were identified in western-lake surface waters at levels comparable to average values reported in studies within Lake Michigan, the North Atlantic Ocean, and the South Pacific Ocean. This study provides insight into the magnitude of microplastic pollution in western Lake Superior, and describes in detail methodology to improve future microplastics studies in aquatic systems.

Arthur James McComb 1936–2017

Professor Arthur McComb conducted pioneer research on the occurrence and mode of action of the plant growth hormones gibberellins for fifteen years. He then applied his experimental skills and physiological knowledge to develop a whole ecosystem approach to the study of aquatic systems. He was passionate in wanting to improve the state of environmental management, based on rational, logical and well-founded biological principles. He and his team focused primarily on the mechanisms controlling plant growth and productivity in aquatic environments, and especially the effects of nutrient enrichment and its consequences, eutrophication. He became a leader in nutrient analysis of water systems, with innovations in how to determine nutrient pathways into waterways and strategies for fixing these issues. This important research has informed the long-term management of several important aquatic systems in Western Australia: the Blackwood River Estuary, the Peel Harvey Estuary, and the protection of seagrasses in Shark Bay, the Swan River and Cockburn Sound. Arthur McComb had a seminal influence on a generation of researchers. Thirty-nine students completed their higher degrees under his supervision and they are spread internationally and throughout Australia in universities, state government departments and consulting firms, confirming his influence on driving our understanding and management of marine, estuarine and freshwater systems.

Xeropreservation of functionalized lipid biomarkers in hyperarid soils in the Atacama Desert

Our understanding of long-term organic matter preservation comes mostly from studies in aquatic systems. In contrast, taphonomic processes in extremely dry environments are relatively understudied and are poorly understood. We investigated the accumulation and preservation of lipid biomarkers in hyperarid soils in the Yungay region of the Atacama Desert. Lipids from seven soil horizons in a 2.5m vertical profile were extracted and analyzed using GC–MS and LC–MS. Diagnostic functionalized lipids and geolipids were detected and increased in abundance and diversity with depth. Deeper clay units contain fossil organic matter (radiocarbon dead) that has been protected from rainwater since the onset of hyperaridity. We show that these clay units contain lipids in an excellent state of structural preservation with functional groups and unsaturated bonds in carbon chains. This indicates that minimal degradation of lipids has occurred in these soils since the time of their deposition between>40,000 and 2million years ago. The exceptional structural preservation of biomarkers is likely due to the long-term hyperaridity that has minimized microbial and enzymatic activity, a taphonomic process we term xeropreservation (i.e., preservation by drying). The degree of biomarker preservation allowed us to reconstruct major changes in ecology in the Yungay region that reflect a shift in hydrological regime from wet to dry since the early Quaternary. Our results suggest that hyperarid environments, which comprise 7.5% of the continental landmass, could represent a rich and relatively unexplored source of paleobiological information on Earth.

Transport distance of invertebrate environmental DNA in a natural river.

Environmental DNA (eDNA) monitoring is a novel molecular technique to detect species in natural habitats. Many eDNA studies in aquatic systems have focused on lake or ponds, and/or on large vertebrate species, but applications to invertebrates in river systems are emerging. A challenge in applying eDNA monitoring in flowing waters is that a species' DNA can be transported downstream. Whether and how far eDNA can be detected due to downstream transport remains largely unknown. In this study we tested for downstream detection of eDNA for two invertebrate species, Daphnia longispina and Unio tumidus, which are lake dwelling species in our study area. The goal was to determine how far away from the source population in a lake their eDNA could be detected in an outflowing river. We sampled water from eleven river sites in regular intervals up to 12.3 km downstream of the lake, developed new eDNA probes for both species, and used a standard PCR and Sanger sequencing detection method to confirm presence of each species' eDNA in the river. We detected D. longispina at all locations and across two time points (July and October); whereas with U. tumidus, we observed a decreased detection rate and did not detect its eDNA after 9.1 km. We also observed a difference in detection for this species at different times of year. The observed movement of eDNA from the source amounting to nearly 10 km for these species indicates that the resolution of an eDNA sample can be large in river systems. Our results indicate that there may be species' specific transport distances for eDNA and demonstrate for the first time that invertebrate eDNA can persist over relatively large distances in a natural river system.

Characterization of the small heat shock protein Hsp27 gene in Chironomus riparius (Diptera) and its expression profile in response to temperature changes and xenobiotic exposures

Small heat shock proteins constitute the most diverse and least conserved group within the large family of heat shock proteins, which play a crucial role in cell response to environmental insults. Chironomus riparius larvae are widely used in environmental research for testing pollutant toxicity in sediments and freshwater environments. Different genes, such as Hsp70, Hsc70, Hsp90, and Hsp40, have been identified in this species as sensitive biomarkers for xenobiotics, but small Hsps genes remain largely unknown. In this study, the Hsp27 has been characterized in C. riparius and its transcriptional response evaluated under several environmental stimuli. The Hsp27 gene was mapped by FISH on polytene chromosomes at region I-C4 and was found to encode a 195 aa protein, which contains an α-crystallin domain bounded by three conserved regions. This protein shows homology with Drosophila melanogaster HSP27, Ceratitis capitata HSP27, and Sarcophaga crassipalpis HSP25. Real-time reverse transcriptase-polymerase chain reaction analysis showed that heat shock (35°C) and cadmium dramatically upregulate this gene. Moreover, exposures to triclosan and bisphenol A were able to significantly increase mRNA levels. However, neither nonylphenol nor tributyltin altered Hsp27 gene expression. The transcriptional activity of Hsp27 gene was modulated during cold stress. Interestingly, cold shock (4°C) significantly reduced Hsp27 transcripts, but this gene was significantly overexpressed during the recovery time at the normal growing temperature. These results show that the Hsp27 gene is sensitive to different environmental stimuli, including endocrine-disrupting pollutants, suggesting its potential as a suitable biomarker for ecotoxicological studies in aquatic systems.

Diet‐tissue fractionation of δ15N by consumers from streams and rivers

Variation in the diet‐tissue fractionation of stable nitrogen isotopes (Δ15N) is a major source of uncertainty in mixing model outputs and the calculation of trophic level in food web studies in aquatic systems. Using δ15N and δ13C of algae and consumers collected from a broad range of streams and rivers in Australia and New Guinea, we calculated Δ15N using a gradient approach, and compared these estimates with those from the literature. Riverine invertebrates and fishes from this region have δ15N diet‐tissue fractionation at the low end of the range of values reported for literature summaries, but different trophic groups had different Δ15N estimates. Source‐consumer regressions based on δ13C had lower slope estimates and lower R2 values compared with those based on δ15N. This implies that although consumers on average obtain a portion of their organic carbon from higher plants, they derive most of their organic nitrogen from N‐rich algae. Use of appropriate Δ15N for riverine consumers will lead to more satisfactory results with isotope mixing models, and the uncertainty associated with Δ15N estimates can be incorporated in the latest iterations of these models that are probabilistic rather than deterministic.

Nutrient stoichiometry and concentrations influence silver toxicity in the aquatic macrophyte Lemna gibba

Though nutrients and silver often co-occur in aquatic ecosystems, the combined effects of these environmental stressors on aquatic plants are poorly understood. Such coexposures are important because nanosilver is increasingly released to the environment, and recent studies in aquatic systems indicate that nanosilver represents an environmental source of ionic silver (Ag+), which exerts relatively high acute toxicity to aquatic life. The primary objective of this study was to understand the effects of nitrogen (N) and phosphorus (P) concentrations and N:P ratios on the toxicity of ionic silver to the model aquatic macrophyte Lemna gibba over 7-d study periods. L. gibba were more sensitive to silver (e.g., lower EC50 values) when N and P concentrations were higher. In addition, greater ionic silver toxicity occurred under higher P-availability (e.g., lower N:P ratios). L. gibba frond number and fresh weight were also differentially affected across nutrient×silver treatment combinations. Such observations highlight the importance of considering site-specific nutrient conditions during the prospective and retrospective risk assessments and management of silver impacts to primary producers.

Determination of free CO2 in emergent groundwaters using a commercial beverage carbonation meter

Summary Dissolved CO2 in groundwater is frequently supersaturated relative to its equilibrium with atmospheric partial pressure and will degas when it is conveyed to the surface. Estimates of dissolved CO2 concentrations can vary widely between different hydrochemical facies because they have different sources of error (e.g., rapid degassing, low alkalinity, non-carbonate alkalinity). We sampled 60 natural spring and mine waters using a beverage industry carbonation meter, which measures dissolved CO2 based on temperature and pressure changes as the sample volume is expanded. Using a modified field protocol, the meter was found to be highly accurate in the range 0.2–35 mM CO2. The meter provided rapid, accurate and precise measurements of dissolved CO2 in natural waters for a range of hydrochemical facies. Dissolved CO2 concentrations measured in the field with the carbonation meter were similar to CO2 determined using the pH-alkalinity approach, but provided immediate results and avoided errors from alkalinity and pH determination. The portability and ease of use of the carbonation meter in the field made it well-suited to sampling in difficult terrain. The carbonation meter has proven useful in the study of aquatic systems where CO2 degassing drives geochemical changes that result in surficial mineral precipitation and deposition, such as tufa, travertine and mine drainage deposits.

The prominence of and biases in biodiversity and ecosystem functioning research

The sub-discipline of biodiversity and ecosystem functioning (BEF) has emerged as a central topic in contemporary ecological research. However, to date no study has evaluated the prominence and publication biases in BEF research. Herein we report the results of a careful quantitative assessment of BEF research published in five core general ecology journals from 1990 to 2007 to determine the position of BEF research within ecology, identify patterns of research effort within BEF research, and discuss their probable proximal and historical causes. The relative importance of BEF publications increased exponentially during the period analyzed and was significantly greater than the average growth of ecological literature, affirming the prominence of BEF as a current paradigm in ecology. However, BEF research exhibited a strong bias toward experimental studies on terrestrial plant communities, with significantly lower effort devoted to the functional aspects of biodiversity in aquatic systems, multiple trophic level systems, and animal or microbial communities. Such trends may be explained by a combination of methodological adequacy and historic epistemological differences in ecological thinking. We suggest that BEF researchers should direct more effort toward the study of aquatic systems and animal communities, emphasize long-term and trophically complex experiments, such as those with multi-trophic microbial communities, employ larger-scale field observational studies and increase the use of integrative and theoretical studies. Many technical and analytical methodologies that are already employed in ecological research, such as stable isotopes, paleobiology, remote sensing, and model selection criteria, can facilitate these aims. Overcoming the above-mentioned shortcomings of current BEF research will greatly improve our ability to predict how biodiversity loss will affect ecosystem processes and services in natural ecosystems.

Nutrients stimulate leaf breakdown rates and detritivore biomass: bottom-up effects via heterotrophic pathways

Most nutrient enrichment studies in aquatic systems have focused on autotrophic food webs in systems where primary producers dominate the resource base. We tested the heterotrophic response to long-term nutrient enrichment in a forested, headwater stream. Our study design consisted of 2 years of pretreatment data in a reference and treatment stream and 2 years of continuous nitrogen (N) + phosphorus addition to the treatment stream. Studies were conducted with two leaf species that differed in initial C:N, Rhododendron maximum (rhododendron) and Acer rubrum (red maple). We determined the effects of nutrient addition on detrital resources (leaf breakdown rates, litter C:N and microbial activity) and tested whether nutrient enrichment affected macroinvertebrate consumers via increased biomass. Leaf breakdown rates were ca. 1.5 and 3x faster during the first and second years of enrichment, respectively, in the treatment stream for both leaf types. Microbial respiration rates of both leaf types were 3x higher with enrichment, and macroinvertebrate biomass associated with leaves increased ca. 2-3x with enrichment. The mass of N in macroinvertebrate biomass relative to leaves tended to increase with enrichment up to 6x for red maple and up to 44x for rhododendron leaves. Lower quality (higher C:N) rhododendron leaves exhibited greater changes in leaf nutrient content and macroinvertebrate response to nutrient enrichment than red maple leaves, suggesting a unique response by different leaf species to nutrient enrichment. Nutrient concentrations used in this study were moderate and equivalent to those in streams draining watersheds with altered land use. Thus, our results suggest that similarly moderate levels of enrichment may affect detrital resource quality and subsequently lead to altered energy and nutrient flow in detrital food webs.

A new automated setup for stable isotope analysis of dissolved organic carbon

The measurement of stable isotope ratios on dissolved organic carbon (DOC) has long posed analytical problems and limited the use of this powerful tracer in biogeochemical studies in aquatic systems. Here, we provide a detailed description of a successful coupling of a custom-modified total organic carbon analyzer (Thermo HiPerTOC) to an isotope-ratio mass spectrometer (IRMS). The method is based on the wet oxidation of up to ~20 mL aqueous sample in a closed reactor, whereby complete oxidation is ensured by a combination of sodium persulfate addition, heating, and UV irradiation. The produced CO2 is carried over a water trap and purification column in a stream of He and introduced into the IRMS via a conventional open-split interface, enabling both quantification and δ13C analysis. Typical reproducibility of δ13C analyses on DOC in this setup is in the order of 0.2‰ or better, comparable to that obtained in a more conventional elemental analyzer-IRMS setup. With appropriate blank correction procedures, accurate analyses can be obtained on concentrations as low as 0.5 mg DOC L−1, representing the lower limit typically observed in marine systems. Typical overall system blank values for DOC analysis are in the order of 1 µg C. Analytical conditions (reaction time, reagent concentrations) were optimized for samples from brackish and marine environments, so that a single method can handle all types of environmental DOC samples. Although no certified DOC standards exist for δ13C, we analyzed the δ13C values of a DOC “consensus reference material” from a deep-ocean environment (cfr. Hansell 2005) and found a δ13C value of −19.5 ± 0.4‰ (n = 3), which is consistent with its oceanic origin.