Spatial and phylogenetic structure of Alpine stonefly assemblages across seven habitats using DNA-species.

Stream ecosystems are spatially heterogeneous, with many different habitat patches distributed within a small area. The influence of this heterogeneity on the biodiversity of benthic insect communities is well documented; however, studies of the role of habitat heterogeneity in species coexistence and assembly remain limited. Here, we investigated how habitat heterogeneity influences spatial structure (beta biodiversity) and phylogenetic structure (evolutionary processes) of benthic stonefly (Plecoptera, Insecta) communities. We sampled 20 sites along two Alpine rivers, including seven habitats in four different reaches (headwaters, meandering, bar-braided floodplain, and lowland spring-fed). We identified 21 morphological species and delineated 52 DNA-species based on sequences from mitochondrial cox1 and nuclear ITS markers. Using DNA-species, we first analysed the patterns of variation in richness, diversity, and assemblage composition by quantifing the contribution of each reach and habitat to the overall DNA-species diversity using an additive partition analysis and distance-based redundancy analysis. Using gene-tree phylogenies, we assessed whether environmental filtering could lead to the co-occurrence of DNA-species using a two-step analysis to detect a phylogenetic signal. All four reaches significantly contributed to DNA-species richness, with the meandering reach having the highest contribution. Habitats had an effect on DNA-species diversity, where glide, riffle and, pool influenced the spatial structure of stonefly assemblage possibly due to the high habitat heterogeneity. Among the habitats, the pool showed significant phylogenetic clustering, suggesting high levels of evolutionary adaptation and strong habitat filtering. This assemblage structure may be caused by long-term stability of the habitat and the similar requirements for co-occurring species. Our study shows the importance of different habitats for the spatial and phylogenetic structure of stonefly assemblage and sheds light on the habitat-specific diversity that may help improve conservation practices.

Local flow convergence, bed scour, and aquatic habitat formation during floods around wooden training structures placed on sand-gravel bars.

Riverine ponds, which are formed and sustained through sediment erosion and deposition, are key habitats for enhancing biodiversity in river reaches. The objective of this study was to understand the roles of traditional river-training wooden structures called "seigyu" on the formation of ponds on nonvegetated bars. Here, the spatial and temporal patterns of the flow and bedform coupled with pond formations for several flood events were assessed. The surface flow patterns were monitored by an unmanned aerial vehicle (UAV) and evaluated by large-scale particle image velocimetry (LSPIV); the maximum flow velocities were 1.3 m s-1 and 1.9 m s-1 during floods when seigyu units were partially and fully submerged, respectively. Although the overall mean flow velocity was greater for the latter events, the spatial variation in flow velocity and dissipation rate of turbulent kinetic energy (TKE) around seigyu was greater for the former events. Such flow patterns affected both bed formation and ecological habitats; ponds were formed at locations beside and behind seigyu, where the flow converged and bed scouring occurred during floods. The frequency and size of ponds around seigyu increased in the early half of the season, and they decreased in the other half when floods were greater in magnitude, which suggests that the bed scouring effect of seigyu was greater in flood stages with partial than fully submergence. Although the bar ponds lack shade to temper the effects of incident light and atmospheric conditions, the ponds displayed smaller daily oscillations in temperature than did the main river, probably due to hyporheic water supply to the ponds. Because many aquatic species cannot tolerate extremely high temperatures in summer, the generation and maintenance of deeper ponds by scouring with sufficient water exchange with the hyporheic zone can be key to enhancing colonization by various aquatic species.

Projection of invertebrate populations in the headwater streams of a temperate catchment under a changing climate.

Climate change places considerable stress on riverine ecosystems by altering flow regimes and increasing water temperature. This study evaluated how water temperature increases under climate change scenarios will affect stream invertebrates in pristine headwater streams. The studied headwater-stream sites were distributed within a temperate catchment of Japan and had similar hydraulic-geographical conditions, but were subject to varying temperature conditions due to altitudinal differences (100 to 850 m). We adopted eight general circulation models (GCMs) to project air temperature under conservative (RCP2.6), intermediate (RCP4.5), and extreme climate scenarios (RCP8.5) during the near (2031-2050) and far (2081-2100) future. Using the water temperature of headwater streams computed by a distributed hydrological-thermal model as a predictor variable, we projected the population density of stream invertebrates in the future scenarios based on generalized linear models. The mean decrease in the temporally averaged population density of Plecoptera was 61.3% among the GCMs, even under RCP2.6 in the near future, whereas density deteriorated even further (90.7%) under RCP8.5 in the far future. Trichoptera density was also projected to greatly deteriorate under RCP8.5 in the far future. We defined taxa that exhibited temperature-sensitive declines under climate change as cold stenotherms and found that most Plecoptera taxa were cold stenotherms in comparison to other orders. Specifically, the taxonomic families that only distribute in Palearctic realm (e.g., Megarcys ochracea and Scopura longa) were selectively assigned, suggesting that Plecoptera family with its restricted distribution in the Palearctic might be a sensitive indicator of climate change. Plecoptera and Trichoptera populations in the headwaters are expected/anticipated to decrease over the considerable geographical range of the catchment, even under the RCP2.6 in the near future. Given headwater invertebrates play important roles in streams, such as contributing to watershed productivity, our results provide useful information for managing streams at the catchment-level.

Evaluation of wastewater nitrogen transformation in a natural wetland (Ulaanbaatar, Mongolia) using dual-isotope analysis of nitrate.

The Tuul River, which provides water for the daily needs of many residents of Ulaanbaatar, Mongolia, has been increasingly polluted by wastewater from the city's sewage treatment plant. Information on water movement and the transformation of water-borne materials is required to alleviate the deterioration of water quality. We conducted a synoptic survey of general water movement, water quality including inorganic nitrogen concentrations, and isotopic composition of nitrogen (δ(15)N-NO(3)(-), δ(18)O-NO(3)(-), and δ(15)N-NH(4)(+)) and water (δ(18)O-H(2)O) in a wetland area that receives wastewater before it enters the Tuul River. We sampled surface water, groundwater, and spring water along the two major water routes in the wetland that flow from the drain of the sewage treatment plant to the Tuul River: a continuous tributary and a discontinuous tributary. The continuous tributary had high ammonium (NH(4)(+)) concentrations and nearly stable δ(15)N-NH(4)(+), δ(15)N-NO(3)(-), and δ(18)O-NO(3)(-) concentrations throughout its length, indicating that nitrogen transformation (i.e., nitrification and denitrification) during transit was small. In contrast, NH(4)(+) concentrations decreased along the discontinuous tributary and nitrate (NO(3)(-)) concentrations were low at many points. Values of δ(15)N-NH(4)(+), δ(15)N-NO(3)(-), and δ(18)O-NO(3)(-) increased with flow along the discontinuous route. Our results indicate that nitrification and denitrification contribute to nitrogen removal in the wetland area along the discontinuous tributary with slow water transport. Differences in hydrological pathways and the velocity of wastewater transport through the wetland area greatly affect the extent of nitrogen removal.

Biodilution of heavy metals in a stream macroinvertebrate food web: evidence from stable isotope analysis.

Analysis of carbon (delta13C) and nitrogen (delta15N) stable isotopes provides an increasingly important means of understanding the complex trophic structure of macroinvertebrate communities in streams. We coupled a stable isotope approach with a contaminant analysis of six metals (Pb, Ag, Zn, Hg, Cu, As) to trace the accumulation and dilution of metals from an abandoned mine across trophic levels of the benthic community in Ginzan Creek, Japan. The delta15N signature increased with trophic level, with mean increases of 4.70 per thousand from producers to primary consumers and 3.06 per thousand from primary to secondary consumers. Tissue Pb and Ag concentrations were negatively correlated with delta15N, indicating biodilution of both metals through the food web. Although macroinvertebrate taxon body mass was negatively correlated with tissue metal concentration at several sites, it did not increase with trophic level (as delta15N) in any of the sites, suggesting that changes in body mass were not the cause of biodilution. Our findings suggest invertebrates at higher trophic levels may exhibit increasingly efficient excretion of metals. Autotrophic epilithon (mean delta13C= -21.3 per thousand) had a much higher concentration of mined metals than did riparian vegetation (mean delta13C= -29.3 per thousand); nonetheless, a carbon-mixing model indicated that taxa feeding on autochthonous carbon sources did not accumulate more metal than allochthonous feeders. It is likely that the notably high metal concentration of allochthonous FPOM plays an important role in the trophic transfer of metals. Our data suggest the strong potential for stable isotope analysis to enhance our understanding of metal transfer through stream macroinvertebrate food webs.

Isolation of sperm vesicles from adult male mayflies and other insects to prepare high molecular weight genomic DNA samples.

We describe here a simple and efficient protocol for genomic DNA isolation from adult males of insects: e.g., Ephemeroptera, Odonata, Orthoptera and Dictyoptera. To minimize contamination of external DNA source, the sperm vesicles were isolated from male individuals from which high molecular weight genomic DNA was extracted. According to this protocol, the genomic DNA samples obtained were high quality (intact), and abundant enough for genotyping analyses and molecular cloning. The protocol reported here enables us to process a huge number of individuals at a time with escaping from cross-contamination, and thus it is quite useful for conducting genetic studies at least in some species of insects. The large yield of high molecular weight DNA from single individual may be advantageous for non PCR-based experiments. As a case study of the protocol, partial coding sequences of histone H3 and EF-1alpha genes are determined for some insects with PCR-amplified DNA fragments.