Alexander Feckler

Inorganic fungicides as routinely applied in organic and conventional agriculture can increase palatability but reduce microbial decomposition of leaf litter

Summary The application of fungicides is considered an indispensable measure to secure crop production. These substances, however, may unintentionally enter surface waters via run-off, potentially affecting the microbial community. To assess such risks adequately, authorities recently called for suitable test designs involving relevant aquatic micro-organisms. We assessed the structural and functional responses of leaf-associated microbial communities, which play a key role in the breakdown of allochthonous leaf material in streams, towards the inorganic fungicides copper (Cu) and elemental sulphur (S). These substances are of particular interest as they are authorized for both conventional and organic farming in many countries of the world. We used the food choice of the amphipod shredder Gammarus fossarum (indicative for micro-organism-mediated leaf palatability) as well as microbial leaf decomposition as functional endpoints. Moreover, the leaf-associated microbial communities were characterized by means of bacterial density, fungal biomass and community composition facilitating mechanistic understanding of the observed functional effects. While Gammarus preferred Cu-exposed leaves over unexposed ones, microbial leaf decomposition was reduced by both Cu and S (up to 30%). Furthermore, Cu exposure decreased bacterial densities (up to 60%), stimulated the growth of leaf-associated fungi (up to 100%) and altered fungal community composition, while S did not affect any of the assessed structural endpoints. Synthesis and applications. We observed both structural and functional changes in leaf-associated microbial communities at inorganic fungicide concentrations realistic for surface water bodies influenced by conventional and organic farming. Our data hence justify a careful re-evaluation of the environmental safety of the agricultural use of these compounds. Moreover, inclusion of an experimental design similar to the one used in this study in lower tier environmental risk assessments of antimicrobial compounds may aid to safeguard the integrity of aquatic microbial communities and the functions they provide.

Impacts of Contaminants on the Ecological Role of Lotic Biofilms

Biofilms play a fundamental ecological role in freshwater ecosystems as they contribute to ecosystem function(s) such as autotrophic primary production, organic matter decomposition and the bottom-up directed energy transfer in the food web. The present focused review summarizes the scientific knowledge on how the roles of autotrophic and heterotrophic biofilms can be modulated as a response to chemical (i.e., pesticide) stress. We discuss how horizontal effects (alterations in the structure of biofilms) can affect the physiological fitness and life history strategy of the next trophic level (vertical effects), namely primary consumers. Since the literature indicates that heterotrophic biofilms are currently at risk as a result of pesticide stress, the protectiveness of environmental risk assessment in Europe as well as North America is questioned. By briefly outlining substantial knowledge gaps, we provide ideas on how the identified uncertainties may be empirically addressed.

When significance becomes insignificant: Effect sizes and their uncertainties in Bayesian and frequentist frameworks as an alternative approach when analyzing ecotoxicological data

Bayesian methods and frequentist confidence intervals are proposed as an alternative approach in ecotoxicology, emphasizing effect sizes and associated (un)certainties to judge the biological relevance of effects instead of basing decisions on p values. These approaches show advantages over null hypothesis significance testing. In particular, Bayesian methods revealed more potential than frequentist counterparts, as the posterior distribution and its credible intervals can be directly interpreted as the probability of effect sizes. Environ Toxicol Chem 2018;37:1949-1955.

Repeated pulse exposures to lambda-cyhalothrin affect the behavior, physiology, and survival of the damselfly larvae Ischnura graellsii (Insecta; Odonata)

Damselflies form an essential part of the aquatic and terrestrial food web. Pesticides may, however, negatively affect their behavior, physiology, and survival. To assess this, a 42-day-lasting bioassay was conducted, during which damselfly larvae (Ischnura graellsii; n = 20) were repeatedly exposed to lambda-cyhalothrin (3 days at; 0, 10, 50, 250, 1250, and 6250ng LCH L-1), followed by recovery phases (4 days) in pesticide-free medium for six weeks. This exposure design was used to simulate frequent runoff events in the field. Variables related to the behavior (strikes against prey and capture success), growth, physiology (lipid content and fatty acid composition), as well as mortality were assessed throughout the experiment. The two highest LCH concentrations induced 100% mortality within the first 48h, whereas 85% of the test organisms survived 28 days under control conditions. The number of strikes against prey was not affected by LCH. In contrast, prey capture success decreased significantly (up to ~50% at 250ng LCH L-1, for instance, after the third pulse exposure) following LCH-exposures compared to the control. This difference was not observed after recovery phases, however, which did not counteract the enhanced energy demand for detoxification and defense mechanisms indicated by a lower growth rate (up to ~20%) and lipid content (up to ~30%) of damselflies at 50 and 250ng LCH L-1. In addition, two essential fatty acids (eicosapentaenoic acid and arachidonic acid) and two precursors (linolenic acid and α-linolenic acid) decreased in their concentrations upon exposure towards 250ng LCH L-1. Thus the results of this study indicate that long-term exposure towards LCH pulses can affect damselfly behavior, physiology and survival. Given the essential role of damselflies in food web dynamics, these effects may potentially translate into local population impairments with subsequent bottom-up directed effects within and across ecosystem boundaries.

Assessing the effects of field-relevant pesticide mixtures for their compliance with the concentration addition model – An experimental approach with Daphnia magna

The environmental risk assessment of pesticides is mainly performed on individual active ingredients. In surface waters within the agricultural landscape, however, contamination is usually characterized by complex pesticide mixtures. To estimate the joint effects caused by these complex mixtures, mathematical models have been proposed. Among these, the model of concentration addition (CA) is suggested as default model for the risk assessment of chemical mixtures as it is considered protective for mixtures composed of similar and dissimilar acting substances. Here we assessed the suitability of CA predictions for seven field relevant pesticide mixtures using acute (immobility) and chronic (reproduction) responses of the standard test species Daphnia magna. Pesticide mixtures indicated largely additive or less than additive effects when using CA model predictions as a reference. Moreover, we revealed that deviations from CA predictions are lower for chronic (up to 3.2-fold) relative to acute (up to 7.2-fold) response variables. Additionally, CA predictions were in general more accurate for complex mixtures relative to those composed of only a few pesticides. Thus, this study suggests CA models as largely protective for the risk assessment of pesticide mixtures justifying its use as default model. At the same time, extrapolating conclusions about the joint effects of pesticides from acute to chronic responses is uncertain, due to partly large discrepancies with regards to the deviation of model prediction and observed effects between exposure scenarios.

Blinded by the light: Increased chlorophyll fluorescence of herbicide-exposed periphyton masks unfavorable structural responses during exposure and recovery

In surface waters within agricultural catchments, periphyton - i.e., biofilms containing algae, heterotrophs, and associated detritus - is subjected to multiple stressors including herbicides. Although herbicide effects on periphyton are frequently studied, the focus has been on photosynthesis-inhibiting herbicides while other modes of toxic action have received little attention. Against this background, a 21-days-lasting bioassay was conducted, during which mature periphytic communities were exposed to the carotenoid-biosynthesis-inhibiting herbicide diflufenican for 12 days (up to 10 μg/L; n = 4), followed by a 9-days-lasting recovery phase in herbicide-free medium. Variables related to periphytic functioning (photosynthetic efficiency and non-photochemical quenching) and structure (pigment concentrations, biomass, and algal community structure) were quantified every third day during both experimental phases. Exposure to ≥ 0.2 μg diflufenican/L resulted in 20-25% and 25-30% lowered carotenoid and chlorophyll a concentrations, respectively, likely explained by a reduced algal biovolume as well as diflufenicans mode of toxic action and thus a shift towards a higher heterotrophy of the communities. Despite these adverse effects on the photosynthetic apparatus, the photosynthetic efficiency increased by up to ∼15% under diflufenican exposure judged on higher chlorophyll fluorescence. This may be explained by an up to ∼60% reduced non-photochemical quenching as well as binding of diflufenican to the pigment-protein membrane complex of the photosystem II, two processes causing higher chlorophyll fluorescence. Additionally, phototrophs may have actively increased energy assimilation to cope with higher energy demands under chemical stress. Although periphyton showed some recovery potential following the exposure phase, observed as increasing chlorophyll a concentrations and non-photochemical quenching, periphyton may not be able to quickly recover from stress given the persistent increase in the photosynthetic efficiency. While the processes underlying the observed effects yet remain speculative, the results suggest a shift towards a higher degree of heterotrophy in periphytic communities ultimately increasing the importance of heterotrophic ecosystem functions at impacted sites over the long term.

Quantitative real-time PCR as a promising tool for the detection and quantification of leaf-associated fungal species – A proof-of-concept using Alatospora pulchella

Traditional methods to identify aquatic hyphomycetes rely on the morphology of released conidia, which can lead to misidentifications or underestimates of species richness due to convergent morphological evolution and the presence of non-sporulating mycelia. Molecular methods allow fungal identification irrespective of the presence of conidia or their morphology. As a proof-of-concept, we established a quantitative real-time polymerase chain reaction (qPCR) assay to accurately quantify the amount of DNA as a proxy for the biomass of an aquatic hyphomycete species (Alatospora pulchella). Our study showed discrimination even among genetically closely-related species, with a high sensitivity and a reliable quantification down to 9.9 fg DNA (3 PCR forming units; LoD) and 155.0 fg DNA (47 PCR forming units; LoQ), respectively. The assay’s specificity was validated for environmental samples that harboured diverse microbial communities and likely contained PCR-inhibiting substances. This makes qPCR a promising tool to gain deeper insights into the ecological roles of aquatic hyphomycetes and other microorganisms.