The Plasticene era: Current uncertainties in estimates of the hazards posed by tiny plastic particles on soils and terrestrial invertebrates.

Plastics are ubiquitous in our daily life. Large quantities of plastics leak in the environment where they weather and fragment into micro- and nanoparticles. This potentially releases additives, but rarely leads to a complete mineralization, thus constitutes an environmental hazard. Plastic pollution in agricultural soils currently represents a major challenge: quantitative data of nanoplastics in soils as well as their effects on biodiversity and ecosystem functions need more attention. Plastic accumulation interferes with soil functions, including water dynamics, aeration, microbial activities, and nutrient cycling processes, thus impairing agricultural crop yield. Plastic debris directly affects living organisms but also acts as contaminant vectors in the soils, increasing the effects and the threats on biodiversity. Finally, the effects of plastics on terrestrial invertebrates, representing major taxa in abundance and diversity in the soil compartment, need urgently more investigation from the infra-individual to the ecosystem scales.

Insights into the molecular mechanisms of pesticide tolerance in the Aporrectodea caliginosa earthworm.

Diffuse pollution of the environment by pesticides has become a major soil threat to non-target organisms, such as earthworms for which declines have been reported. However some endogeic species are still abundant and persist in intensively cultivated fields, suggesting they become tolerant to long-term anthropogenic pressure. We thus considered the working hypothesis that populations of Aporrectodea caliginosa earthworms from conventionally managed fields developed a tolerance to pesticides compared with those from organically managed fields. To investigate this hypothesis, we studied earthworm populations of the same genetic lineage from soils that were either lowly or highly contaminated by pesticides to detect any constitutive expression of differentially expressed molecular pathways between these populations. Earthworm populations were then experimentally exposed to a fungicide-epoxiconazole-in the laboratory to identify different molecular responses when newly exposed to a pesticide. State-of-the-art omics technology (RNA sequencing) and bioinformatics were used to characterize molecular mechanisms of tolerance in a non-targeted way. Additional physiological traits (respirometry, growth, bioaccumulation) were monitored to assess tolerance at higher levels of biological organization. In the present study, we generated the de novo assembly transcriptome of A. caliginosa consisting of 64,556 contigs with N50 = 2862 pb. In total, 43,569 Gene Ontology terms were identified for 21,593 annotated sequences under the three main ontologies (biological processes, cellular components and molecular functions). Overall, we revealed that two same lineage populations of A. caliginosa earthworms, inhabiting similar pedo-climatic environment, have distinct gene expression pathways after they long-lived in differently managed agricultural soils with a contrasted pesticide exposure history for more than 22 years. The main difference was observed regarding metabolism, with upregulated pathways linked to proteolytic activities and the mitochondrial respiratory chain in the highly exposed population. This study improves our understanding of the long-term impact of chronic exposure of soil engineers to pesticide residues.

Is the Cyanobacterial Bloom Composition Shifting Due to Climate Forcing or Nutrient Changes? Example of a Shallow Eutrophic Reservoir.

Cyanobacterial blooms in eutrophic freshwater is a global threat to the functioning of ecosystems, human health and the economy. Parties responsible for the ecosystems and human health increasingly demand reliable predictions of cyanobacterial development to support necessary decisions. Long-term data series help with identifying environmental drivers of cyanobacterial developments in the context of climatic and anthropogenic pressure. Here, we analyzed 13 years of eutrophication and climatic data of a shallow temperate reservoir showing a high interannual variability of cyanobacterial development and composition, which is a less occurring and/or less described phenomenon compared to recurrant monospecific blooms. While between 2007-2012 Planktothrix agardhii dominated the cyanobacterial community, it shifted towards Microcystis sp. and then Dolichospermum sp. afterwards (2013-2019). The shift to Microcystis sp. dominance was mainly influenced by generally calmer and warmer conditions. The later shift to Dolichospermum sp. was driven by droughts influencing, amongst others, the N-load, as P remained unchanged over the time period. Both, climatic pressure and N-limitation contributed to the high variability of cyanobacterial blooms and may lead to a new equilibrium. The further reduction of P-load in parallel to the decreasing N-load is important to suppress cyanobacterial blooms and ameliorate ecosystem health.

Thermal plasticity and sensitivity to insecticides in populations of an invasive beetle: Cyfluthrin increases vulnerability to extreme temperature.

Climate change increases average temperatures and the occurrence of extreme weather events, in turn accentuating the risk of organism exposure to temperature stress. When thermal conditions become stressful, the sensitivity of insects toward insecticides can be exacerbated. Likewise, exposure of insects to insecticides can subsequently influence their ability to handle stressful temperatures. Here, we investigated the effects of constant temperature and daily heat spikes, in presence/absence of insecticide treatment (cyfluthrin), on the condition (impairment of mobility) and thermal tolerance to cold (-6 °C) and heat (42.5 °C) of the terrestrial beetle Alphitobius diaperinus. The responses of insects from four populations (three farm-collected populations, one laboratory population) to different durations of extreme temperature exposure were compared. The results showed that the laboratory population was generally more sensitive to extreme cold and heat temperatures, with less than 50% of adults recovering after an exposure at -6 or +42.5 °C for 3h. Significant differences in the level of thermal tolerance were also found among insects from poultry farms. Cyfluthrin exposure incurred detrimental effects to insects' condition in all but one population. For two out of the four populations, mobility impairment was increased when adults were exposed to daily heat spikes (6 h per day at 38 °C) and cyfluthrin simultaneously, compared to cyfluthrin exposure at constant temperatures; yet, no significant interaction between the two stressors was found. Finally, using one farm collected population, effects of pre-exposure to cyfluthrin on extreme temperature tolerance provided another example of the toxicant-induced climate sensitivity in insects.

Cell free Microcystis aeruginosa spent medium affects Daphnia magna survival and stress response.

Primary consumers in freshwater ecosystems, such as the zooplankton organism Daphnia magna, are highly affected by cyanobacteria, both as they may use it as a food source but also by cyanobacterial metabolites present in the water. Here, we investigate the impacts of cyanobacterial metabolites focussing on the environmental realistic scenario of the naturally released mixture without crushing cyanobacterial cells or their uptake as food. Therefore, D. magna were exposed to two concentrations of cell free cyanobacterial spent medium from Microcystis aeruginosa PCC 7806 to represent higher and lower ecologically-relevant concentrations of cyanobacterial metabolites. Including microcystin-LR, 11 metabolites have been detected of which 5 were quantified. Hypothesising concentration and time dependent negative impact, survival, gene expression marking digestion and metabolism, oxidative stress response, cell cycle and molting as well as activities of detoxification and antioxidant enzymes were followed for 7 days. D. magna suffered from oxidative stress as both catalase and glutathione S-transferase enzyme activities significantly decreased, suggesting enzyme exhaustibility after 3 and 7 days. Moreover, gene-expressions of the 4 stress markers (glutathione S-transferase, glutathione peroxidase, catalase and thioredoxin) were merely downregulated after 7 days of exposure. Energy allocation (expression of glyceraldehyde-3-phosphate dehydrogenase) was increased after 3 days but decreased as well after 7 days exposure. Cell cycle was impacted time dependently but differently by the two concentrations, along with an increasing downregulation of myosin heavy chain responsible for cell arrangement and muscular movements. Deregulation of nuclear hormone receptor genes indicate that D. magna hormonal steering including molting seemed impaired despite no detection of microviridin J in the extracts. As a consequence of all those responses and presumably of more than investigated molecular and physiological changes, D. magna survival was impaired over time, in a concentration dependent manner. Our results confirm that besides microcystin-LR, other secondary metabolites contribute to negative impact on D. magna survival and stress response.

Cross talk: Two way allelopathic interactions between toxic Microcystis and Daphnia.

Due to eutrophication, freshwater ecosystems frequently experience cyanobacterial blooms, many of which produce bioactive metabolites that can affect vertebrates and invertebrates life traits. Zooplankton are able to develop tolerance as a physiological response to cyanobacteria and their bioactive compounds, however, this comes with energetic cost that in turn influence Daphnia life traits and may impair populations. Vice versa, it has been suggested that Daphnia are able to reduce cyanobacterial dominance until a certain cyanobacterial density; it remains unclear whether Daphnia metabolites alone influence the physiological state and bioactive metabolites production of cyanobacteria. Hence, this study investigates mutual physiological reactions of toxic Microcystis aeruginosa PCC7806 and Daphnia magna. We hypothesize that a) the presence of D. magna will negatively affect growth, increase stress response and metabolites production in M. aeruginosa PCC7806 and b) the presence of M. aeruginosa PCC7806 will negatively affect physiological responses and life traits in D. magna. In order to test these hypotheses experiments were conducted in a specially designed co-culture chamber that allows exchange of the metabolites without direct contact. A clear mutual impact was evidenced. Cyanobacterial metabolites reduced survival of D. magna and decreased oxidative stress enzyme activity. Simultaneously, presence of D. magna did not affect photosynthetic activity. However, ROS increase and tendencies in cell density decrease were observed on the same day, suggesting possible energy allocation towards anti-oxidative stress enzymes, or other protection mechanisms against Daphnia infochemicals, as the strain managed to recover. Elevated concentration of intracellular and overall extracellular microcystin MC-LR, as well as intracellular concentrations of aerucyclamide A and D in the presence of Daphnia, indicating a potential protective or anti-grazing function. However, more research is needed to confirm these findings.

Transdisciplinary Bioblitz: Rapid biotic and abiotic inventory allows studying environmental changes over 60 years at the Biological Field Station of Paimpont (Brittany, France) and opens new interdisciplinary research opportunities.

BACKGROUND: The Biological Field Station of Paimpont (Station Biologique de Paimpont, SBP), owned by the University of Rennes and located in the Brocéliande Forest of Brittany (France), has been hosting student scientific research and field trips during the last 60 years. The study area of the SBP is a landscape mosaic of 17 ha composed of gorse moors, forests, prairies, ponds and creeks. Land use has evolved over time. Historical surveys by students and researchers focused on insects and birds. With this study, we aimed to increase the range of taxa observations, document changes in species composition and landscape and provide a basis for interdisciplinary research perspectives. We gathered historical data, implemented an all-taxon biodiversity inventory (ATBI) in different habitats of the SBP study area, measured abiotic factors in the air, water and soil and performed a photographical landscape observation during the BioBlitz held in July 2017. NEW INFORMATION: During the 24 h BioBlitz, organised by the SBP and the EcoBio lab from the University of Rennes and the French National Center of Scientific Research (CNRS), different habitats were individually sampled. Seventy-seven experts, accompanied by 120 citizens and 12 young people participating in the European Volunteer Service, observed, identified and databased 660 species covering 5 kingdoms, 8 phyla, 21 classes, 90 orders and 247 families. In total, there were 1819 occurrences including records identified to higher taxon ranks, thereby adding one more kingdom and four more phyla. Historical data collection resulted in 1176 species and 4270 occurrences databased. We also recorded 13 climatic parameters, 10 soil parameters and 18 water parameters during the BioBlitz. Current habitats were mapped and socio-ecological landscape changes were assessed with a diachronic approach using 32 historical photographs and historical maps. The coupling of historical biodiversity data with new biotic and abiotic data and a photographic comparison of landscape changes allows an integrative understanding of how the SBP changed from agriculturally-used land to a managed natural area within the last 60 years. Hence, this BioBlitz represents an important holistic sampling of biodiversity for studies on trophic webs or on trophic interactions or on very diverse, but connected, habitats. The integration of social, biotic and abiotic data opens innovative research opportunities on the evolution of socio-ecosystems and landscapes.

How can interspecific interactions in freshwater benthic macroinvertebrates modify trace element availability from sediment?

This study aimed to assess how bioturbation by freshwater benthic macroinvertebrates with different biological traits alone or in combination could modify trace elements (TE) fate between sediment and water, and if water TE concentration and animal TE content impair their body stores. Three macroinvertebrate species were exposed to TE contaminated sediment for 7 days: the omnivorous Echinogammarus berilloni (Amphipoda), the sediment feeding Tubifex tubifex (Oligochaeta) and the filter feeding Pisidium sp. (Bivalvia). Treatments were one without invertebrates (control), two with amphipods or mussels alone, and the combinations amphipod-mussel, and amphipod-mussel-worms. Water TE concentration increased significantly in 2 or 3 species mesocosms, concerning mainly Rare Earth Elements, Cr, U and Pb, known to be associated to the colloidal phase. By contrast, water soluble TE were not affected by animals. For both, amphipods and mussels, TE body content increased with the number of coexisting species. For amphipods, this increase concerned both, soluble and colloid-associated TE, possibly due to intense contact and feeding from sediment and predation on tubificids. TE bioaccumulation in mussel was less important and characterized by soluble TE, with water filtration as most plausible uptake route. Protein, triglyceride and Whole Body Energy Budget increased in amphipods with the number of coexisting species (probably by feeding on mussels' feces and tubificids) whereas triglycerides declined in mussels (presumably filtration was disturbed by amphipods). This study highlights interspecific interactions as key drivers explaining both: TE bioturbation, depending on their water solubility or colloidal association, and the exposure/contamination of species through another species activity.

Daphnia magna Exudates Impact Physiological and Metabolic Changes in Microcystis aeruginosa.

While the intracellular function of many toxic and bioactive cyanobacterial metabolites is not yet known, microcystins have been suggested to have a protective role in the cyanobacterial metabolism, giving advantage to toxic over nontoxic strains under stress conditions. The zooplankton grazer Daphnia reduce cyanobacterial dominance until a certain density, which may be supported by Daphnia exudates, affecting the cyanobacterial physiological state and metabolites' production. Therefore, we hypothesized that D. magna spent medium will impact the production of cyanobacterial bioactive metabolites and affect cyanobacterial photosynthetic activity in the nontoxic, but not the toxic strain. Microcystin (MC-LR and des-MC-LR) producing M. aeruginosa PCC7806 and its non-microcystin producing mutant were exposed to spent media of different D. magna densities and culture durations. D. magna spent medium of the highest density (200/L) cultivated for the shortest time (24 h) provoked the strongest effect. D.magna spent medium negatively impacted the photosynthetic activity of M. aeruginosa PCC7806, as well as the dynamics of intracellular and extracellular cyanobacterial metabolites, while its mutant was unaffected. In the presence of Daphnia medium, microcystin does not appear to have a protective role for the strain. On the contrary, extracellular cyanopeptolin A increased in M. aeruginosa PCC7806 although the potential anti-grazing role of this compound would require further studies.

Transgenerational effects of cyanobacterial toxins on a tropical micro-crustacean Daphnia lumholtzi across three generations.

Climate change and human activities induce an increased frequency and intensity of cyanobacterial blooms which could release toxins to aquatic ecosystems. Zooplankton communities belong to the first affected organisms, but in tropical freshwater ecosystems, this issue has yet been poorly investigated. We tested two questions (i) if the tropical Daphnia lumholtzi is capable to develop tolerance to an ecologically relevant concentration of purified microcystin-LR and microcystins from cyanobacterial extract transferable to F1 and F2 generations? And (ii) would F1 and F2 generations recover if reared in toxin-free medium? To answer these questions, we conducted two full factorial mutigenerational experiments, in which D. lumholtzi was exposed to MC-LR and cyanobacterial extract at the concentration of 1 µg L-1 microcystin continuously for three generations. After each generation, each treatment was spit into two: one reared in the control (toxin free) while the other continued in the respective exposure. Fitness-related traits including survival, maturity age, body length, and fecundity of each D. lumholtzi generation were quantified. Though there were only some weak negative effects of the toxins on the first generation (F0), we found strong direct, accumulated and carried-over impacts of the toxins on life history traits of D. lumholtzi on the F1 and F2, including reductions of survival, and reproduction. The maturity age and body length showed some inconsistent patterns between generations and need further investigations. The survival, maturity age (for extract), and body length (for MC-LR) were only recovered when offspring from toxin exposed mothers were raised in clean medium for two generations. Chronic exposure to long lasting blooms, even at low density, evidently reduces survival of D. lumholtzi in tropical lakes and reservoirs with ecological consequences.

Sensitivity of a tropical micro-crustacean (Daphnia lumholtzi) to trace metals tested in natural water of the Mekong River.

Metal contamination is one of the major issues to the environment worldwide, yet it is poorly known how exposure to metals affects tropical species. We assessed the sensitivity of a tropical micro-crustacean Daphnia lumholtzi to three trace metals: copper (Cu), zinc (Zn) and nickel (Ni). Both, acute and chronic toxicity tests were conducted with metals dissolved in in situ water collected from two sites in the lower part of the Mekong River. In the acute toxicity test, D. lumholtzi neonates were exposed to Cu (3-30µgL-1), Zn (50-540µgL-1) or Ni (46-2356µgL-1) for 48h. The values of median lethal concentrations (48h-LC50) were 11.57-16.67µg Cu L-1, 179.3-280.9µg Zn L-1, and 1026-1516µg Ni L-1. In the chronic toxicity test, animals were exposed to Cu (3 and 4µgL-1), Zn (50 and 56µgL-1), and Ni (six concentrations from 5 to 302µgL-1) for 21days. The concentrations of 4µg Cu L-1 and 6µg Ni L-1 enhanced the body length of D. lumholtzi but 46µg Ni L-1 and 50µg Zn L-1 resulted in a strong mortality, reduced the body length, postponed the maturation, and lowered the fecundity. The results tentatively suggest that D. lumholtzi showed a higher sensitivity to metals than related species in the temperate region. The results underscore the importance of including the local species in ecological risk assessment in important tropical ecosystems such as the Mekong River to arrive at a better conservational and management plan and regulatory policy to protect freshwater biodiversity from metal contamination.

Dynamics of cyanobacteria and cyanobacterial toxins and their correlation with environmental parameters in Tri An Reservoir, Vietnam.

This study evaluates the water quality from Tri An Reservoir, a drinking water supply for several million people in southern Vietnam, in terms of cyanobacterial biomass and their potent toxins, microcystins (MCs). Cyanobacteria, their toxins and environmental parameters were monitored monthly for 1 year (April 2008-March 2009) at six stations covering a transect through the reservoir. Dynamics of cyanobacterial abundance in relation to cyanobacterial biomass, toxins and environmental factors were investigated. Environmental variables from Tri An Reservoir favored algal and cyanobacterial development. However, cyanobacterial biomass and proportion varied widely, influenced by physical conditions, available nutrients and nutrient competition among the phytoplankton groups. Cyanobacterial biomass correlated slightly positively to temperature, pH and biochemical oxygen demand (BOD5), but negatively to total inorganic nitrogen concentrations. During most of the sampling times, MC concentrations in the reservoir were quite low (≤0.07 µg L(-1) MC-LR equivalent), and presented a slight positive correlation to BOD5, total nitrogen:total phosphorus ratio and cyanobacterial biomass. However, in cyanobacterial scum samples, which now and then occurred in the reservoir, MC concentrations reached up to 640 µg g(-1) DW(-1). The occurrence of MC in the reservoir poses a risk to local residents who use the water daily for domestic purposes.

Accumulation and detoxication responses of the gastropod Lymnaea stagnalis to single and combined exposures to natural (cyanobacteria) and anthropogenic (the herbicide RoundUp(®) Flash) stressors.

Freshwater gastropods are increasingly exposed to multiple stressors in the field such as the herbicide glyphosate in Roundup formulations and cyanobacterial blooms either producing or not producing microcystins (MCs), potentially leading to interacting effects. Here, the responses of Lymnaea stagnalis to a 21-day exposure to non-MC or MC-producing (33µgL(-1)) Planktothrix agardhii alone or in combination with the commercial formulation RoundUp(®) Flash at a concentration of 1µgL(-1) glyphosate, followed by 14days of depuration, were studied via i) accumulation of free and bound MCs in tissues, and ii) activities of anti-oxidant (catalase CAT) and biotransformation (glutathione-S-transferase GST) enzymes. During the intoxication, the cyanobacterial exposure induced an early increase of CAT activity, independently of the MC content, probably related to the production of secondary cyanobacterial metabolites. The GST activity was induced by RoundUp(®) Flash alone or in combination with non MC-producing cyanobacteria, but was inhibited by MC-producing cyanobacteria with or without RoundUp(®) Flash. Moreover, MC accumulation in L. stagnalis was 3.2 times increased when snails were concomitantly exposed to MC-producing cyanobacteria with RoundUp(®), suggesting interacting effects of MCs on biotransformation processes. The potent inhibition of detoxication systems by MCs and RoundUp(®) Flash was reversible during the depuration, during which CAT and GST activities were significantly higher in snails previously exposed to MC-producing cyanobacteria with or without RoundUp(®) Flash than in other conditions, probably related to the oxidative stress caused by accumulated MCs remaining in tissues.

ß-N-methylamino-L-alanine (BMAA) uptake by the animal model, Daphnia magna and subsequent oxidative stress.

ß-N-methylamino-l-alanine (BMAA), produced by cyanobacteria, is a neurotoxin implicated in Amyotrophic lateral sclerosis/Parkinsonism dementia complex (ALS/PDC). BMAA concentrations in cyanobacteria are lower than those thought to be necessary to result in neurological damage thus bioaccumulation or biomagnification is required to achieve concentrations able to cause neurodegeneration. Many cyanobacteria produce BMAA and uptake routes into the food web require examination. In this study we investigate the uptake of BMAA by adult phytoplanktivorus Daphnia magna via exposure to dissolved pure BMAA and BMAA containing cyanobacteria, as well as the subsequent oxidative stress response in the daphnia. Free BMAA and protein-associated BMAA were quantified by LC-MS/MS. Dissolved BMAA was taken up and was found as free BMAA in D. magna. No protein-associated BMAA was detected in D. magna after a 24-h exposure period. No BMAA was detectable in D. magna after exposure to BMAA containing cyanobacteria. BMAA inhibited the oxidative stress defence and biotransformation enzymes within 24-h exposure in the tested Daphnia and could therefore impair the oxidant status and the capability of detoxifying other substances in D. magna.

Abiotic stressors and stress responses: What commonalities appear between species across biological organization levels?

Organisms are regularly subjected to abiotic stressors related to increasing anthropogenic activities, including chemicals and climatic changes that induce major stresses. Based on various key taxa involved in ecosystem functioning (photosynthetic microorganisms, plants, invertebrates), we review how organisms respond and adapt to chemical- and temperature-induced stresses from molecular to population level. Using field-realistic studies, our integrative analysis aims to compare i) how molecular and physiological mechanisms related to protection, repair and energy allocation can impact life history traits of stressed organisms, and ii) to what extent trait responses influence individual and population responses. Common response mechanisms are evident at molecular and cellular scales but become rather difficult to define at higher levels due to evolutionary distance and environmental complexity. We provide new insights into the understanding of the impact of molecular and cellular responses on individual and population dynamics and assess the potential related effects on communities and ecosystem functioning.

Earthworm tolerance to residual agricultural pesticide contamination: field and experimental assessment of detoxification capabilities.

This study investigates if acclimatization to residual pesticide contamination in agricultural soils is reflected in detoxification, antioxidant enzyme activities and energy budget of earthworms. Five fields within a joint agricultural area exhibited different chemical and farming histories from conventional cultivation to organic pasture. Soil multiresidual pesticide analysis revealed up to 9 molecules including atrazine up to 2.4 ng g(-1) dry soil. Exposure history of endogeic Aporrectodea caliginosa and Allolobophora chlorotica modified their responses to pesticides. In the field, activities of soluble glutathione-S-transferases (sGST) and catalase increased with soil pesticide contamination in A. caliginosa. Pesticide stress was reflected in depletion of energy reserves in A. chlorotica. Acute exposure of pre-adapted and naïve A. caliginosa to pesticides (fungicide Opus(®), 0.1 µg active ingredient epoxiconazole g(-1) dry soil, RoundUp Flash(®), 2.5 µg active ingredient glyphosate g(-1) dry soil, and their mixture), revealed that environmental pre-exposure accelerated activation of the detoxification enzyme sGST towards epoxiconazole.

Specific proteomic response of Unio pictorum mussel to a mixture of glyphosate and microcystin-LR.

Cyanobacterial toxins and pesticides regularly impact freshwaters. Microcystin-LR is one of the most toxic and common cyanobacterial toxins whereas glyphosate is the active ingredient of a widely use herbicide. As filter feeders, freshwater mussels are particularly exposed. Like many native bivalve species, Unio pictorum suffers from a continuous decline in Europe. In order to get a deeper insight of its response to contaminants, U. pictorum was exposed to either 10 µg L(-1) of microcystin-LR or 10 µg L(-1) of glyphosate or a mixture of both. Proteins of the digestive glands were extracted and analyzed by DIGE. Gel analysis revealed 103 spots with statistical variations, and the response seems to be less toward glyphosate than to microcystin-LR. Specific spots have variations only when exposed to the mixture, showing that there is an interaction of both contaminants in the responses triggered. The proteins of 30 spots have been identified. They belong mostly to the cytoskeleton family, but proteins of the oxidative pathway, detoxification, and energetic metabolism were affected either by glyphosate or microcystin-LR or by the mixture. These results demonstrate the importance to study contaminants at low concentrations representative of those found in the field and that multicontaminations can lead to different response pathways.

Transgenerational effects of microcystin-LR on Daphnia magna.

Anthropogenic and climate factors increase the frequency of problematic cyanobacterial blooms in freshwater. Among other toxins, some cyanobacteria produce microcystins (MCs), which inhibit phosphatases type 1 and type 2A and provokes oxidative stress. Toxic cyanobacteria affect the growth, survival and reproduction of zooplankton, particularly those from the genus Daphnia, which have a central position in pelagic food webs. However, one possibility to ameliorate effects is to biotransform MC via glutathione S transferase (GST) to a less toxic glutathione conjugate. This process was hypothesised to underlie the ability of Daphnia to withstand MC and to explain the enhanced tolerance of the offspring from mothers exposed to toxic cyanobacteria. Thus we conducted multigenerational experiments with D. magna, exposing the parental generation to MC for 1 or 7 days and determining the enzyme-mediated tolerance to MC in their offspring by assessing the acute effect of MC on biotransformation and antioxidant and metabolism enzymes, and through 21 day chronic tests on toxicity and growth. Seven days of exposure of the parental generation to MC induced higher activity of GST and malate dehydrogenase in the offspring and enabled them to increment the catalase activity when challenged with MC, whereas 1 day of exposure of the parental generation did not. Offspring from non-exposed and 1-day-exposed mothers suffered decreased survival when exposed to MC compared with offspring from 7-day-exposed mothers; survival was correlated with the elevated activity of GST, malate dehydrogenase and catalase, suggesting maternal transfer of activation factors. However, increased survival occurred at the expense of individual growth. These results suggest that transgenerational effects are provoked by MC in D. magna, which may explain the observed acquirement of enhanced tolerance over generations.

Adaptation of freshwater mussels to cyanobacterial toxins: response of the biotransformation and antioxidant enzymes.

Freshwater mussels such as the invasive Dreissena polymorpha and the indigenous Unio tumidus nourish by high filtration rates and may accumulate cyanobacteria and their toxins during cyanobacterial blooms. Physiological adaptations to cyanotoxins enable organisms to endure cyanobacterial blooms but may differ between species. Biotransformation and excretion capacities for cyanobacteria and anthropogenic pollutants have been demonstrated for Dreissena polymorpha but less for unionid species. This study compares the activities of biotransformation (glutathione S-transferase, GST) and antioxidant enzymes (superoxide dismutase, SOD and catalase, CAT) in Dreissena polymorpha to Unio tumidus in response to cyanotoxin exposure (10 µg L(-1) and 50 µg L(-1) microcystin-LR, respectively, total microcystin from a cyanobacterial crude extract) for 24 h and 7d exposure duration. Enzyme activities in Dreissena polymorpha were measured in the whole mussel tissue, digestive gland and in gills and in Unio tumidus in the digestive gland, gills, mantle, foot as well as in the remaining tissue. The sGST was elevated for the entire exposure period in the whole mussel tissue of Dreissena polymorpha but despite higher basal activities in digestive gland and gills of Unio tumidus, it was rather inhibited or unaltered in most of their tissues. Elevated SOD activity indicated oxidative stress response in Dreissena polymorpha, but not in Unio tumidus. The CAT activity was barely affected in both species, rather inhibited in Unio tumidus, despite again higher basal activities in digestive gland and remaining tissue. Compared to the indigenous Unio tumidus, the investigated biotransformation and oxidative stress combating enzymes respond stronger in the invasive Dreissena polymorpha.

The synthetic gestagen Levonorgestrel disrupts sexual development in Xenopus laevis by affecting gene expression of pituitary gonadotropins and gonadal steroidogenic enzymes.

In the present study, Xenopus laevis tadpoles were chronically exposed to four concentrations of the synthetic gestagen Levonorgestrel (LNG; 10(-11), 10(-10), 10(-9), and 10(-8)M) starting at Nieuwkoop and Faber (NF) stage 48 until completion of metamorphosis. At NF 58 and 66, brain-pituitary and gonad samples were taken for gene expression analyses of gonadotropins and gonadal steroidogenic enzymes. Exposure to 10(-9) and 10(-8)M LNG until NF 58 repressed messenger RNA (mRNA) expression of luteinizing hormone (LH) ß in both genders. This decrease was persistent after further treatment until NF 66 in the 10(-8)M LNG treatment. Expression of follicle-stimulating hormone (FSH) ß was affected sex-specifically. No effect was present in NF 58 females, whereas LNG at 10(-9) and 10(-8)M significantly increased FSHß mRNA levels in males. In NF 66 females, 10(-9)M LNG treatment increased FSHß gene expression, whereas a decrease was observed in NF 66 males exposed to 10(-8)M LNG. In gonads, expression of steroid-5-alpha-reductase was affected sex-specifically with increased mRNA levels in females but repressed levels in males. Gene expression of further gonadal steroidogenic factors was decreased by 10(-8)M LNG in both genders at NF 66. Assessment of gonad gross morphology and histology revealed poorly developed testes in the 10(-8)M LNG treatment. Our results reveal considerable effects of chronic LNG exposure on sexual development of amphibians. The persistent inhibition of LHß expression concomitant with decreased mRNA levels of gonadal steroidogenic enzymes is suggested to result in the disruption of reproduction in adult amphibians.