Organic-matter decomposition as a bioassessment tool of stream functioning: A comparison of eight decomposition-based indicators exposed to different environmental changes.

Organic-matter decomposition has long been proposed as a tool to assess stream functional integrity, but this indicator largely depends on organic-matter selection. We assessed eight decomposition-based indicators along two well-known environmental gradients, a nutrient-enrichment gradient (0.2-1.4 mg DIN/L) in central Portugal and an acidification gradient (pH: 4.69-7.33) in north-eastern France to identify the most effective organic-matter indicator for assessing stream functional integrity. Functional indicators included natural leaf litter (alder and oak) in 10-mm and 0.5-mm mesh bags, commercial tea (Lipton green and rooibos teas in 0.25-mm mesh bags), wood sticks (wood tongue depressors) and cotton strips. Biotic indices based on benthic macroinvertebrates (IPtIN for Portugal and IBGN for France) were calculated to compare the effectiveness of structural and functional indicators in detecting stream impairment and to assess the relationship between both types of indicators. The effectiveness of organic-matter decomposition rates as a functional indicator depended on the stressor considered and the substrate used. Decomposition rates generally identified nutrient enrichment and acidification in the most acidic streams. Decomposition rates of alder and oak leaves in coarse-mesh bags, green and rooibos teas and wood sticks were positively related with pH. Only decomposition rates of rooibos tea and wood sticks were related with DIN concentration; decomposition rates along the nutrient-enrichment gradient were confounded by differences in shredder abundance and temperature among streams. Stream structural integrity was good to excellent across streams; the IPtIN index was unrelated to DIN concentration, while the IBGN index was positively related with pH. The relationships between decomposition rates and biotic indices were loose in most cases, and only decomposition rates of alder leaves in coarse-mesh bags and green tea were positively related with the IBGN. Commercial substrates may be a good alternative to leaf litter to assess stream functional integrity, especially in the case of nutrient enrichment.

Metabolomic, functional, and ecologic responses of the common freshwater fungus Neonectria lugdunensis to mine drainage stress.

Metal contamination of watersheds is a global problem. Here, we conducted litter decomposition studies with Neonectria lugdunensis, a cosmopolitan aquatic fungus. Fungal isolates from four reference (non-impacted) and six metal-contaminated streams (due to mine drainage) were exposed to mine drainage and reference stream waters in Central Portugal. Impact of mine drainage waters on N. lugdunensis hyphae was investigated by performing metabolomic profiling of 200 lipids and 25 amino acids (AA) with ultra-high performance liquid chromatography-mass spectrometry. In parallel, functional response of N. lugdunensis isolates was assessed through expression profiles of a functional gene, cellobiohydrolase I (CbhI). Ecological performance via leaf mass loss was also determined. Exposure to mine drainage waters altered the concentration of numerous AA and lipids. Most strikingly, a gradual increase in the concentration of the triacylglycerols (TAG) with shorter acyl chains and lesser unsaturation was observed after the exposure to mine drainage waters. In addition, the changes in the concentration of numerous TAG, lysophosphatidylcholines, and AA were more significant in the isolates from the metal-contaminated streams after exposure to mine drainage water. CbhI gene of the isolates from reference streams was down-regulated by metal stress, while those from metal-contaminated streams remained unaffected. Finally, leaf mass loss was influenced by both exposure to mine drainage waters and the origin of isolates. Overall, our study demonstrates unique functional signatures displayed by fungi under metal stress and the relevant role that fungal AA and lipids play to cope with metal toxicity.

Biodiversity of leaf litter fungi in streams along a latitudinal gradient.

Global patterns of biodiversity have emerged for soil microorganisms, plants and animals, and the extraordinary significance of microbial functions in ecosystems is also well established. Virtually unknown, however, are large-scale patterns of microbial diversity in freshwaters, although these aquatic ecosystems are hotspots of biodiversity and biogeochemical processes. Here we report on the first large-scale study of biodiversity of leaf-litter fungi in streams along a latitudinal gradient unravelled by Illumina sequencing. The study is based on fungal communities colonizing standardized plant litter in 19 globally distributed stream locations between 69°N and 44°S. Fungal richness suggests a hump-shaped distribution along the latitudinal gradient. Strikingly, community composition of fungi was more clearly related to thermal preferences than to biogeography. Our results suggest that identifying differences in key environmental drivers, such as temperature, among taxa and ecosystem types is critical to unravel the global patterns of aquatic fungal diversity.

Uranium toxicity to aquatic invertebrates: A laboratory assay.

Uranium mining is an environmental concern because of runoff and the potential for toxic effects on the biota. To investigate uranium toxicity to freshwater invertebrates, we conducted a 96-h acute toxicity test to determine lethal concentrations (testing concentrations up to 262 mg L-1) for three stream invertebrates: a shredder caddisfly, Schizopelex festiva Rambur (Trichoptera, Sericostomatidae); a detritivorous isopod, Proasellus sp. (Isopoda, Asellidae); and a scraper gastropod, Theodoxus fluviatilis (Gastropoda, Neritidae). Next, we ran a chronic-toxicity test with the most tolerant species (S. festiva) to assess if uranium concentrations found in some local streams (up to 25 µg L-1) affect feeding, growth and respiration rates. Finally, we investigated whether S. festiva takes up uranium from the water and/or from ingested food. In the acute test, S. festiva survived in all uranium concentrations tested. LC50-96-h for Proasellus sp and T. fluviatilis were 142 mg L-1 and 24 mg L-1, respectively. Specimens of S. festiva exposed to 25 µg L-1 had 47% reduced growth compared with specimens under control conditions (21.5 ±â€¯2.9 vs. 40.6 ±â€¯4.9 µg of mass increase animal-1·day-1). Respiration rates (0.40 ±â€¯0.03 µg O2·h-1·mg animal-1) and consumption rates (0.54 ±â€¯0.05 µg µg animal-1·day-1; means ±â€¯SE) did not differ between treatments. Under laboratory conditions S. festiva accumulated uranium from both the water and the ingested food. Our results indicate that uranium can be less toxic than other metals or metalloids produced by mining activities. However, even at the low concentrations observed in streams affected by abandoned mines, uranium can impair physiological processes, is bioaccumulated, and is potentially transferred through food webs.

Could contaminant induced mutations lead to a genetic diversity overestimation?

Contaminant driven genetic erosion reported through the inspection of selectable traits can be underestimated using neutral markers. This divergence was previously reported in the aquatic system of an abandoned pyrite mine. The most sensitive genotypes of the microcrustacean cladoceran Daphnia longispina were found to be lacking in the impacted reservoir near the entrance of the metal rich acid mine drainage (AMD). Since that divergence could be, at least partially, accounted for by mutagenicity and genotoxicity of the AMD, the present study aimed at providing such a characterization. The Allium cepa chromosomal aberration assay, using root meristematic cells, was carried out, by exposing seeds to 100, 10, 1, and 0.1 % of the local AMD. Chromosomal aberrations, cell division phases and cell death were quantified after the AMD exposure and after 24 and 48 h recovery periods. The AMD revealed to be mutagenic and genotoxic, even after diluting it to 1 and 0.1 %. Dilutions within this range were previously found to be below the lethality threshold and to elicit sublethal effects on reproduction of locally collected D. longispina clonal lineages Significant mutagenic effects (micronuclei and chromosomal breaks) were also found at 0.1 % AMD, supporting that exposure may induce permanent genetic alterations. Recovery tests showed that AMD genotoxic effects persisted after the exposure.

An in situ bioassay integrating individual and biochemical responses using small fish species.

The interest in the ecological relevance of risk assessments and, thus, in in situ bioassays has been increasing in the last years. The present study developed a time- and cost-effective in situ bioassay, aiming at obtaining, in a short period of time and with a minimum of resources, a set of ecologically relevant toxicological information in a site-specific approach. Poecilia reticulata and Gambusia holbrooki were chosen as test species. Post-exposure feeding inhibition and the biomarkers acetylcholinesterase, lactate dehydrogenase and glutathione S-transferases were the endpoints tested. The battery of biomarkers as a whole was sensitive to the in situ exposure in an acid mine drainage impacted effluent, although responses varied between test species. Post-exposure feeding inhibition was the most sensitive endpoint, and its association with biomarker responses was discussed. The linkage between individual responses, such as feeding, and biomarkers suggested that, at least in this case, biomarkers can be relevant at higher levels of biological organization. Altogether, the proposed short-term in situ bioassay seems to be a promising tool, since it represents a reasonable compromise between sensitivity, time/cost-effectiveness and ecological relevance.