• Energy Research

Fungi, particularly aquatic hyphomycetes, have been recognized as playing a dominant role in microbial decomposition of plant litter in streams. In this study, we used a microcosm experiment with different levels of fungal diversity (species number and identity) using monocultures and combinations with up to five aquatic hyphomycete species (Articulospora tetracladia, Tricladium splendens, Heliscus submersus, Tetrachaetum elegans and Flagellospora curta) to assess the effects of ethanol and phenanthrene on three functional measures: plant litter decomposition, fungal biomass accrual and reproduction. Alder leaves were conditioned by fungi for 7days and then were exposed to phenanthrene (1mgL(-1)) dissolved in ethanol (0.1% final concentration) or ethanol (at the concentration used to solubilise phenanthrene) for further 24days. Exposure to ethanol alone or in combination with phenanthrene decreased leaf decomposition and fungal reproduction, but increased fungal biomass produced. All aspects of fungal activity varied with species number. Fungal activity in polycultures was generally higher than that expected from the sum of the weighted performances of participating species in monoculture, suggesting complementarity between species. However, the activity of fungi in polycultures did not exceed the activity of the most productive species either in the absence or presence of ethanol alone or with phenanthrene.

Invasive alien species (IAS) can disrupt important ecological functions in aquatic ecosystems; however, many of these effects are not quantified and remain speculative. In this study, we assessed the effects of the invasive crayfish Procambarus clarkii (Girard, 1852) on leaf litter decomposition (a key ecosystem process) and associated invertebrates using laboratory and field manipulative experiments. The crayfish had significant impacts on leaf decomposition due to direct consumption of leaf litter and production of fine particulate organic matter, and indirectly due to consumption of invertebrate shredders. The invertebrate community did not appear to recognize P. clarkii as a predator, at least in the first stages after its introduction in the system; but this situation might change with time. Overall, results suggested that the omnivore invader P. clarkii has the potential to affect detritus-based food webs through consumption of basal resources (leaf litter) and/or consumers. Recognizing that this IAS is widespread in Europe, Asia and Africa, and may attain high density and biomass in aquatic ecosystems, our results are important to develop strategies for improving stream ecosystem functioning and to support management actions aiming to control the invasive omnivore P. clarkii.

We conducted a microcosm experiment with monocultures and all possible combinations of four aquatic hyphomycete species, Articulospora tetracladia, Flagellospora curta, Geniculospora grandis and Heliscus submersus, to examine the potential effects of species richness on three functional aspects: leaf litter decomposition (leaf mass loss), fungal production (ergosterol buildup) and reproductive effort (released spores). Both species richness and identity significantly affected fungal biomass and conidial production (number and biomass of released spores), whereas only species identity had a significant effect on leaf mass loss. In mixed cultures, all measures of fungal functions were greater than expected from the weighted performances of participating species in monoculture. Mixed cultures outperformed the most active monoculture for biomass accumulation but not for leaf mass loss and conidial production. The three examined aspects of aquatic hyphomycete activity tended to increase with species richness, and a complementary effect was unequivocally demonstrated for fungal biomass. Our results also suggest that specific traits of certain species may have a greater influence on ecosystem functioning than species number.

Microorganisms are key drivers of leaf litter decomposition; however, the mechanisms underlying the dynamics of different microbial groups are poorly understood. We investigated the effects of seasonal variation and invertebrates on fungal and bacterial dynamics, and on leaf litter decomposition. We followed the decomposition of Populus nigra litter in a Mediterranean stream through an annual cycle, using fine and coarse mesh bags. Irrespective of the season, microbial decomposition followed two stages. Initially, bacterial contribution to total microbial biomass was higher compared to later stages, and it was related to disaccharide and lignin degradation; in a later stage, bacteria were less important and were associated with hemicellulose and cellulose degradation, while fungi were related to lignin decomposition. The relevance of microbial groups in decomposition differed among seasons: fungi were more important in spring, whereas in summer, water quality changes seemed to favour bacteria and slowed down lignin and hemicellulose degradation. Invertebrates influenced litter-associated microbial assemblages (especially bacteria), stimulated enzyme efficiencies and reduced fungal biomass. We conclude that bacterial and fungal assemblages play distinctive roles in microbial decomposition and differ in their sensitivity to environmental changes, ultimately affecting litter decomposition, which might be particularly relevant in highly seasonal ecosystems, such as intermittent streams.

Nanocopper oxide (nanoCuO) is among the most widely used metal oxide nanoparticles which increases their chance of being released into freshwaters. Fungi are the major microbial decomposers of plant litter in streams. Fungal laccases are multicopper oxidase enzymes that are involved in the degradation of lignin and various xenobiotic compounds. We investigated the effects of nanoCuO (5 levels, ≤ 200 mg L(-1)) on four fungal isolates collected from metal-polluted and non-polluted streams by analyzing biomass production, changes in mycelial morphology, laccase activity, and quantifying copper adsorbed to mycelia, and ionic and nanoparticulate copper in the growth media. The exposure to nanoCuO decreased the biomass produced by all fungi in a concentration- and time-dependent manner. Inhibition of biomass production was stronger in fungi from non-polluted (EC₅₀(10 days) ≤ 31 mg L(-1)) than from metal-polluted streams (EC₅₀(10 days) ≥ 65.2 mg L(-1)). NanoCuO exposure led to cell shrinkage and mycelial degeneration, particularly in fungi collected from non-polluted streams. Adsorption of nanoCuO to fungal mycelia increased with the concentration of nanoCuO in the medium and was higher in fungi from non-polluted streams. Extracellular laccase activity was induced by nanoCuO in two fungal isolates in a concentration-dependent manner, and was highly correlated with adsorbed Cu and/or ionic Cu released by dissolution from nanoCuO. Putative laccase gene fragments were also detected in these fungi. Lack of substantial laccase activity in the other fungal isolates was corroborated by the absence of laccase-like gene fragments.

We investigated how a community of microbial decomposers adapted to a reference site responds to a sudden decrease in the water quality. For that, we assessed the activity and diversity of fungi and bacteria on decomposing leaves that were transplanted from a reference (E1) to a polluted site (E2), and results were compared to those from decomposing leaves either at E1 or E2. The two sites had contrasting concentrations of organic and inorganic nutrients and heavy metals in the stream water. At E2, leaf decomposition rates, fungal biomass, and sporulation were reduced, while bacterial biomass was stimulated. Fungal diversity was four times lower at the polluted site. The structure of fungal community on leaves decomposing at E2 significantly differed from that decomposing at E1, as indicated by the principal response curves analysis. Articulospora tetracladia, Anguillospora filiformis, and Lunulospora curvula were dominant species on leaves decomposing at E1 and were the most negatively affected by the transfer to the polluted site. The transfer of leaves colonized at the reference site to the polluted site reduced fungal diversity and sporulation but not fungal biomass and leaf decomposition. Overall, results suggest that the high diversity on leaves from the upstream site might have mitigated the impact of anthropogenic stress on microbial decomposition of leaves transplanted to the polluted site.

Urbanization and industrial activities have contributed to widespread contamination by metals and polycyclic aromatic hydrocarbons, but the combined effects of these toxics on aquatic biota and processes are poorly understood. We examined the effects of cadmium (Cd) and phenanthrene on the activity and diversity of fungi associated with decomposing leaf litter in streams. Leaves of Alnus glutinosa were immersed for 10 days in an unpolluted low-order stream in northwest Portugal to allow microbial colonization. Leaves were then exposed in microcosms for 14 days to Cd (0.06-4.5 mg L(-1)) and phenanthrene (0.2 mg L(-1)) either alone or in mixture. A total of 19 aquatic hyphomycete species were found sporulating on leaves during the whole study. The dominant species was Articulospora tetracladia, followed by Alatospora pulchella, Clavatospora longibrachiata, and Tetrachaetum elegans. Exposure to Cd and phenanthrene decreased the contribution of A. tetracladia to the total conidial production, whereas it increased that of A. pulchella. Fungal diversity, assessed as denaturing gradient gel electrophoresis fingerprinting or conidial morphology, was decreased by the exposure to Cd and/or phenanthrene. Moreover, increased Cd concentrations decreased leaf decomposition and fungal reproduction but did not inhibit fungal biomass production. Exposure to phenanthrene potentiated the negative effects of Cd on fungal diversity and activity, suggesting that the co-occurrence of these stressors may pose additional risk to aquatic biodiversity and stream ecosystem functioning.

SummaryA consequence of drought in streams is the emersion of decomposing leaf litter, which may alter organic matter recycling. We assessed the effects of emersion on decomposition of black poplar leaves and associated microbes (microbial biomass, extracellular enzyme activities and microbial diversity) in two streams with distinct characteristics, in particular nutrients, temperature and oxygen levels. Leaf decomposition rates, fungal biomass and extracellular enzyme activities were lower in the most impacted stream (high nutrients and temperature, low oxygen). Also, the structure of fungal and bacterial communities differed between streams. Emersion strongly affected all microbial functional measures. Leaf decomposition, fungal biomass and extracellular enzyme activities were more sensitive at the most pristine site, while fungal reproduction and bacterial biomass production were more affected by emersion at the most impacted stream. Microbial community structure was strongly altered after emersion. Although similar effects on leaf‐associated microbes were found in both streams, functional responses to emersion differed probably as a consequence of different initial microbial communities with different sensitivity to the drying stress. Our study highlights the need of understanding the effects of drought in streams suffering from different environmental perturbations, since responses to emersion appear to depend on the environmental context.