Uptake and physiological effects of the neonicotinoid imidacloprid and its commercial formulation Confidor® in a widespread freshwater oligochaete.

The neonicotinoid imidacloprid (IMI) is one of the most extensively applied neuro-active insecticides worldwide and continues to enter surface waters in many countries despite a recent ban for outdoor use in the EU. Yet little is known about ecotoxicological effects on non-target benthic freshwater species exposed to environmentally relevant concentrations of IMI and its marketed products. The aim of the present study was to narrow this gap by assessing effects of pure IMI and its commercial formulation Confidor® on the aquatic oligochaete Lumbriculus variegatus, a key species in freshwater sediments. To this end, we determined dose-response relationships in 24 h toxicity tests, bioconcentration during 24 h and 5 d of exposure to 0.1, 1 and 10 µg IMI L-1, and physiological stress responses by measuring glutathione S-transferase, glutathione reductase and catalase activity in the same conditions. Maximum neonicotinoid concentrations reported from the field were lethal to L. variegatus within 24 h (LC50 of 65 and 88 µg IMI L-1 in pure form and as active ingredient of Confidor®, respectively). At sub-lethal exposure concentrations, tissue content of IMI significantly increased with exposure time. The observed bioconcentration factors (BCFs) were far above the water octanol coefficient (KOW), indicating a potentially large underestimation of IMI bioaccumulation when based on KOW. Activities of biotransformation and antioxidant enzymes indicated attempts of L. variegatus to counter xenobiotic-triggered oxidative stress to very low IMI and Confidor® concentrations. Together, our data add significantly to growing evidence that the continued proliferation of neonicotinoids require increased efforts in environmental risk assessment, especially in view of species-specific differences in sensitivities to the insecticide and possibly to additives of commercial formulations.

Protein association of ß-N-methylamino-L-alanine in Triticum aestivum via irrigation.

Bioaccumulation of several cyanotoxins has been observed in numerous food webs. More recently, the neurotoxic, non-proteinogenic amino acid ß-N-methylamino-L-alanine (BMAA) was shown to biomagnify in marine food webs. It was thus necessary to assess whether a human exposure risk via a terrestrial food source could exist. As shown for other cyanotoxins, spray irrigation of crop plants with cyanobacterial bloom-contaminated surface water poses the risk of toxin transfer into edible plant parts. Therefore, in the present study, we evaluated a possible transfer of BMAA via spray irrigation into the seeds of one of the world's most widely cultivated crop plants, Triticum aestivum. Wheat plants were irrigated with water containing 10 µg L-1 BMAA until they reached maturity and seed-bearing stage (205 days). Several morphological characteristics, such as germination rate, number of roots per seedling, length of primary root and cotyledon, and diameter of the stems were evaluated to assess the effects of chronic exposure. After 205 days, BMAA bioaccumulation was quantified in roots, shoots, and mature seeds of T. aestivum. No adverse morphology effects were observed and no free intracellular BMAA was detected in any of the exposed plants. However, in mature seeds, protein-associated BMAA was detected at 217 ± 150 ng g FW-1; significantly more than in roots and shoots. This result demonstrates the unexpected bioaccumulation of a hydrophilic compound and highlights the demand to specify in addition to limit values for drinking water, tolerable daily intake rates for the cyanobacterial-neurotoxin BMAA.

Uptake, distribution in different tissues, and genotoxicity of imidacloprid in the freshwater fish Australoheros facetus.

The neonicotinoid imidacloprid is under re-evaluation by regulatory agencies because of the poor current information available regarding its potential effects. One of the goals of the present study was to determine imidacloprid uptake and distribution in the freshwater fish Australoheros facetus experimentally exposed for 24 h and 48 h to 100 µg/L, 300 µg/L, and 2500 µg/L. The toxicity of imidacloprid to fish reported in the literature is in the milligrams per liter or gram per liter range, but sublethal effects at micrograms per liter in some groups other than fish have been described. Another goal of the present study was to evaluate imidacloprid's potential genotoxicity and to compare it between the individual compound and a commercial formulation. Concentrations of imidacloprid were measured in water, brain, muscle, gills, gut, liver, and blood by liquid chromatography-tandem mass spectrometry. Imidacloprid was detected in all the tissues tested. Concentrations were higher after 48 h than after 24 h in liver, gills, gut, and muscle, whereas in brain and blood they were similar at both exposure times. Although there was no accumulation, only uptake, of imidacloprid, genotoxicity was observed. In fish exposed to IMIDA NOVA 35® , increased micronucleus frequency at 100 µg/L and 1000 µg/L was detected, whereas in the imidacloprid active ingredient bioassay it increased only at 1000 µg/L imidacloprid. The present findings warn of the possible consequences that fish living in freshwater ecosystems can suffer. Environ Toxicol Chem 2017;36:699-708. © 2016 SETAC.

Time dependent uptake, bioaccumulation and biotransformation of cell free crude extract microcystins from Lake Amatitlán, Guatemala by Ceratophyllum demersum, Egeria densa and Hydrilla verticillata.

Recent studies evidence that macrophytes can uptake and bioaccumulate microcystins (MC) from contaminated environments, suggesting their use in phytoremediation. In the present study Ceratophyllum demersum, Egeria densa and Hydrilla verticillata were exposed to cell free crude extracts (CE) containing three MC congeners MC-LR, MC-RR and MC-YR at a total MC concentration of 104.4 ± 7.6 µg/L from Lake Amatitlán, Guatemala. Time dependent total glutathione (tGSH), glutathione disulfide (GSSG), disappearance of MC from exposure medium and macrophyte uptake as well as calculated uptake and biotransformation rates and bioconcentration factors (BCF) were monitored after 1, 4, 8 hours (h) and 1, 3, 7 and 14 days (d). Results showed that tGSH concentrations in all exposed macrophytes were enhanced by CE. Disappearance of 62.1 ± 13, 40.8 ± 3.1 and 37.8 ± 3.5 µg/L total MCs from exposure mediums with E. densa, H. verticillata and C. demersum were observed after 1 h. Followed by the total elimination of MCs in exposure medium from H. verticillata after 14 d. Highest MC bioaccumulation capacity (BCF), was observed in E. densa followed by C. demersum and H. verticillata. The here presented results imply the strong MC phytoremediation potential of the evaluated macrophytes.

Single and combined exposure to MC-LR and BMAA confirm suitability of Aegagropila linnaei for use in green liver systems(®)-A case study with cyanobacterial toxins.

The filamentous green algae Aegagropila linnaei was tested for its uptake capacity of the cyanobacterial toxins microcystin-LR (MC-LR) and ß-N-methylamino-l-alanine (BMAA) in order to approve the suitability of its use in the Green Liver System(®). Uptake into the algae and toxin reduction in the medium were analyzed by LC-MS/MS after static exposure for one week to 20µgL(-1) MC-LR, 80µgL(-1) BMAA, and 20µgL(-1) MC-LR together with 80µgL(-1) BMAA, respectively. BMAA was effectively removed by A. linnaei within 5 days compared to only around 35% removal of the initial exposure concentration in the case of MC-LR, independent of the application mode, in single or in a mixture. However, differences were found for BMAA amounts taken up into the tissue in that it was higher if applied in combination with MC-LR. Additionally, physiological responses such as the activity of biotransformation enzyme glutathione S-transferase (GST), antioxidant enzymes peroxidase (POD) and catalase (CAT) as well as the development of the reactive oxygen species hydrogen peroxide (H2O2) were compared between the different treatment groups in order to determine possible harmful effects of the toxin exposure on the algae. In contrast to the toxin exposure to a single toxin with no significant enzymatic response, exposure to the toxin mixture provoked an immediate increase in GST and CAT activity after one day as well as after longer exposure for one week, hinting on an enhanced need for prevention against exposure derived reactive oxygen species as well as putative biotransformation attempts in a mixture exposure scenario.

Antioxidative response of the three macrophytes Ceratophyllum demersum, Egeria densa, and Hydrilla verticillata to a time dependent exposure of cell-free crude extracts containing three microcystins from cyanobacterial blooms of Lake Amatitlán, Guatemala.

Microcystins (MCs) produced by cyanobacteria in natural environments are a potential risk to the integrity of ecosystems. In this study, the effects of cyanobacterial cell-free crude extracts from a Microcystis aeruginosa bloom containing three MC-congeners MC-LR, -RR, and -YR at environmental relevant concentrations of 49.3±2.9, 49.8±5.9, and 6.9±3.8µg/L, respectively, were evaluated on Ceratophyllum demersum (L.), Egeria densa (Planch.), and Hydrilla verticillata (L.f.). Effects on photosynthetic pigments (total chlorophyll (chl), chl a, chl b, and carotenoids), enzymatic defense led by catalase (CAT), peroxidase (POD) and glutathione reductase (GR), and biotransformation enzyme glutathione S-transferase (GST) were measured after 1, 4, and 8h and after 1, 3, 7, and 14 days of exposure. Results show that in all exposed macrophytes, photosynthetic pigments were negatively affected. While chl a and total chl decreased with increasing exposure time, a parallel increase in chl b was observed after 8h. Concomitant increase of ∼5, 16, and 34% of antioxidant carotenoid concentration in exposed C. demersum, E. densa, and H. verticillata, respectively, was also displayed. Enzymatic antioxidant defense systems in all exposed macrophytes were initiated within the first hour of exposure. In exposed E. densa, highest values of CAT and GR activities were observed after 4 and 8h, respectively, while in exposed H. verticillata highest value of POD activity was observed after 8h. An early induction with a significant increase of biotransformation enzyme GST was observed in E. densa after 4h and in C. demersum and H. verticillata after 8h. These results are the first to show rapid induction of stress and further possible MC biotransformation (based on the activation of GST enzymatic activity included in MC metabolization during the biotransformation mechanism) in macrophytes exposed to crude extract containing a mixture of MCs.

Uptake of a cyanotoxin, ß-N-methylamino-L-alanine, by wheat (Triticum aestivum).

In order to study the uptake of the cyanobacterial neurotoxin ß-N-methylamino-l-alanine (BMAA) into the crop plant Triticum aestivum during germination and primary growth imbibed grains and 7-day-old seedlings were irrigated with 100 and 1000µg l(-1) BMAA for 4 days and 100µg l(-1) BMAA for 28 days. Content of derivatized free and protein-associated BMAA in seedlings, root and shoot tissue, respectively, were analyzed by LC-MS/MS. Free BMAA was only detected in seedlings exposed to 1000µg l(-1) BMAA, whereas protein-associated BMAA was found at both exposure concentrations. Irrigation with 100µgl(-1) BMAA led to an uptake of the neurotoxin into roots and shoots and to immediate protein-association. In roots, protein-associated BMAA was detectable after 5 days with peaking amounts after 14 days. Longer exposure did not cause further accumulation in roots. In contrast, protein-associated BMAA was detected in shoot samples after only 1 day. In shoots the highest amounts of protein-associated BMAA were found after 28 days. In turn, in both plant compartments free BMAA was below the measurable concentration.

The fate of the cyanobacterial toxin ß-N-methylamino-L-alanine in freshwater mussels.

The cyanobacterial neurotoxin, ß-N-methylamino-l-alanine (BMAA) has been suggested as a causative agent for certain neurodegenerative diseases. This cyanotoxin bioaccumulates in an array of aquatic organisms, in which it occurs as both a free amino acid and in a protein-associated form. This study was intended to investigate the environmental fate of BMAA by examining the metabolism of isotopically labeled BMAA in four freshwater mussel species. All species showed substantial uptake of BMAA from the culture media. Data showed no significant evidence for BMAA catabolism in any of the animals but did suggest metabolism via the reversible covalent modification of BMAA in freshwater mussels, a process that appears to be variable in different species.

Uptake of the cyanobacterial neurotoxin, anatoxin-a, and alterations in oxidative stress in the submerged aquatic plant Ceratophyllum demersum.

The prevalence of cyanobacterial blooms in fresh water bodies worldwide has become a serious environmental problem. The blooms can increase the occurrence of cyanobacterial neurotoxin, anatoxin-a, and this toxin can interact with aquatic plants and other pivotal components of aquatic ecosystems. Despite this, several questions regarding the uptake of the toxin by aquatic plants and its association with toxic effects still remain. This study investigated the uptake of anatoxin-a in relation to alterations in oxidative stress, estimated by changes in lipid peroxidation and tocopherol contents (alpha-, beta-, gamma-, and delta-tocopherol), in the submerged aquatic plant, Ceratophylum demersum, at environmentally relevant concentrations. Exposure to five different concentrations of anatoxin-a (0.005, 0.05, 0.5, 5 and 50µgl(-1)) for 24h increased concentrations in C. demersum in a dose-dependent manner. All four forms of tocopherols were elevated at low concentrations of anatoxin-a (0.005. 0.05. 0.5 and 5µgl(-1)). However, a decline in the four tocopherol forms along with a high level of lipid peroxidation was observed at 50µgl(-1) exposure dose. During 336-h exposure to 15µgl(-1) anatoxin-a, rapid toxin uptake during the first 24h and subsequent steady accumulation of the toxin were observed. The four tocopherol forms increased in response to anatoxin-a uptake, attaining their maximum levels together with a significant increase of lipid peroxidation after 12 or 24h. After 24-h exposure, the four tocopherol forms decreased gradually without recovery. The results clearly indicate that anatoxin-a uptake can cause a disturbance of the oxidative stress in the aquatic plant, and depending on the concentration and exposure duration, oxidative damage occurs.

Accumulation of microcystin congeners in different aquatic plants and crops--a case study from lake Amatitlán, Guatemala.

Microcystins (MCs) fate in natural environments can lead to its transfer into aquatic organisms, e.g. aquatic plants. Moreover, lakes in several countries sustain agriculture activities posing a serious health threat for the public. The case of Lake Amatitlán in Guatemala, was addressed to better understand MCs accumulation of four aquatic plants (Polygonum portoricensis, Eichhornia crassipes, Typha sp. and Hydrilla verticillata) coexisting with Microcystis aeruginosa blooms. These findings were further corroborated with an uptake/accumulation laboratory study. Finally crop products (Solanum lycopersicum and Capsicum annuum) irrigated with lake water were also evaluated for MCs. The obtained results suggest that Lake Amatitlán is highly contaminated with MCs (intra- and extracellular 1931 and 90 µg/L, respectively). In fruits of S. lycopersicum and C. annuum a concentration of 1.16 and 1.03 µg/kg dry weight (DW), respectively could be detected. All four aquatic plants showed a high MCs uptake capacity based on obtained bioconcentration factors (BCF) 165, 18, 16 and 11, respectively. These results were further corroborated in a laboratory study with 30 percent of total MCs taken up by H. verticillata within just 7 days. Additionally it was evidenced that all plants accumulated more MC-LR than other MCs congeners. Monitoring of crop products irrigated with lake water needs further consideration.

ß-N-Methylamino-L-alanine exposure alters defense against oxidative stress in aquatic plants Lomariopsis lineata, Fontinalis antipyretica, Riccia fluitans and Taxiphyllum barbieri.

Four different aquatic plants, the Pteridophyte Lomariopsis lineata and the Bryophytes Fontinalis antipyretica, Riccia fluitans and Taxiphyllum barbieri, were tested for their capacity to absorb the neurotoxin ß-N-Methylamino-L-alanine (BMAA) from water and thus their possible applicability in a "Green Liver System". After exposure to 10 and 100 µg L(-1) BMAA for 1, 3, 7 and 14 days exposure concentration of medium and tissue were analyzed by LC-MS/MS. The amount removed by the plants within only 1 day was equal to the biological degradation of 14 days. Comparing the "BMAA-removal" capacity of the 4 tested aquatic plants R. fluitans, L. lineata and T. barbieri turned out to be most effective in cleaning the water from this cyanobacterial toxin by up to 97% within 14 days. Activity of the antioxidant enzymes peroxidase (POD) and catalase (CAT), as well as biotransformation enzyme glutathione S-transferase (GST) was compared between exposed and control plants to determine possible harmful effects induced by BMAA. Whereas the Bryophytes displayed increased POD activity and subsequent adaptation when exposed to the lower concentration, as well as partly inhibited antioxidant response at the higher applied BMAA concentration, the Pteridophyte L. lineata reacted with increased POD activity during the whole experiment and increased GST activity after longer exposure for 14 days. To give a recommendation of the suitability of an aquatic plant to be used for sustainable phytoremediation of contaminated water, testing of removal capacity of specific contaminants as well as studying general physiological parameters giving hint on survivability in such environments has to be combined.

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.

Exposure to human pharmaceuticals Carbamazepine, Ibuprofen and Bezafibrate causes molecular effects in Dreissena polymorpha.

Carbamazepine (CBZ), Ibuprofen (IBU) and Bezafibrate (BEZ) were tested for their potential to bioaccumulate and provoke molecular changes in the non-target organism Dreissena polymorpha. mRNA changes of enzymes and other proteins involved in the prevention from protein damage (heat shock protein 70, hsp70) and oxidative stress (superoxide dismutase, SOD; catalase, CAT; metallothionein, MT), biotransformation (pi-class glutathione S-transferase, piGST; aryl hydrocarbon receptor, AH-R), elimination (P-glycoprotein, P-gp) and reversible protein posttranslational modification (protein phosphatase 2A, PP2A) served as molecular biomarkers. Mussels were exposed in a flow-through system to increasing concentrations of the three substances (1, 10, 100 and 1000 nM). The two lower concentrations correspond to environmentally relevant concentrations detected in surface and effluent waters, respectively. Measuring tissue concentration after one, four and seven days the uptake of CBZ and IBU by the mussels could be evidenced, whereas no accumulation data could be achieved for BEZ. The bioconcentration factor was highest for mussels exposed to the lowest CBZ and IBU concentrations, with 90 and 460-fold higher tissue concentration, respectively, after seven days. CBZ was the only substance tested which caused a significant increase in gill mRNA level of hsp70 after only one day exposure, evidencing the potential of CBZ to immediately provoke a stress condition and assumingly protein damage in gills. After longer exposure, mussels displayed down-regulated mRNA levels of hsp70 and SOD in gills, as well as of MT and P-gp in the digestive gland, hinting on an inhibitory character of CBZ. In IBU exposed mussels increased oxidant stress conditions were evidenced by induced mRNA levels in the digestive gland of CAT and MT, as well as SOD after one and four days, respectively. A concentration as found at sewage treatment plant effluents provoked an increase in transcript levels of piGST, suggesting enhanced need for biotransformation of IBU or by-products derived from oxidative stress. Also exposure to an environmentally relevant BEZ concentration provoked an immediate increase in piGST transcript level in the digestive gland followed by up-regulated hsp70 after four and seven days evidencing a chronic stress condition for the mussels.

The synthetic gestagen levonorgestrel impairs metamorphosis in Xenopus laevis by disruption of the thyroid system.

Synthetic gestagens, including levonorgestrel (LNG), are active compounds in contraceptives, and several studies report their occurrence in surface waters. However, information about endocrine-disrupting effects in nontarget organisms is scarce. The present study investigated effects of LNG exposure on thyroid hormone-dependent metamorphosis of Xenopus laevis. Premetamorphic X. laevis tadpoles at Nieuwkoop and Faber (NF) stage 48 were exposed in a flow-through culture system to four LNG concentrations (10(-11), 10(-10), 10(-9), and 10(-8)M) over the period of metamorphosis. At NF 58 and 66, tadpoles were examined sex specifically. Developmental time and organismal responses were recorded and correlated with molecular and histopathological endpoints. Exposure to 10(-8)M LNG caused an inhibition of metamorphosis resulting in developmental arrest at early climax stages as giant tadpoles or tailed frogs. In brain-pituitary tissue of NF 58 tadpoles, gene expression of thyroid-stimulating hormone (ß-subunit; TSHß), TH receptor ß (TRß), and deiodinase type 3 (D3) was not changed. Instead, prolactin (PRL) messenger RNA (mRNA) was significantly increased by 10(-9)M LNG in females and by 10(-8)M LNG in both sexes. In NF 66 tadpoles, mRNA levels of TSHß mRNA were significantly increased in the 10(-9) and 10(-8)M LNG treatment groups indicating a hypothyroid state. No changes of TRß, D3, and PRL gene expression were detected. Histopathological evaluation of thyroid gland sections revealed no typical sign of hypothyroidism but rather an inactivated appearance of the thyroid. In conclusion, our data demonstrate for the first time a completely new aspect of thyroid system disruption caused by synthetic gestagens in developing amphibians.

Molecular effects and bioaccumulation of levonorgestrel in the non-target organism Dreissena polymorpha.

Bioaccumulation and effects of the contraceptive hormone levonorgestrel were examined in the non-target organism Dreissena polymorpha. Molecular biomarkers of biotransformation, elimination, antioxidant defence and protein damage were analyzed after exposure to increasing concentrations of levonorgestrel in a flow-through system. The lowest concentration (0.312 µg L(-1)) was 100-fold bioconcentrated within four days. A decrease of the bioconcentration factor was observed within one week for the highest test concentrations (3.12 and 6.24 µg L(-1)) suggesting enhanced excretory processes. The immediate mRNA up-regulation of pi class glutathione S-transferase proved that phase II biotransformation processes were induced. Disturbance of fundamental cell functions was assumed since the aryl hydrocarbon receptor has been permanently down-regulated. mRNA up-regulation of P-glycoprotein, superoxide dismutase and metallothioneine suggested enhanced elimination processes and ongoing oxidative stress. mRNA up-regulation of heat shock protein 70 in mussels exposed to the two highest concentrations clearly indicated impacts on protein damage.

The beta-receptor blocker metoprolol alters detoxification processes in the non-target organism Dreissena polymorpha.

Due to increasing amounts of pharmaceutically active compounds (PhACs) in the aquatic environment, their largely unknown effects to non-target organisms need to be assessed. This study examined physiological changes in the freshwater mussel Dreissena polymorpha exposed to increasing concentrations (0.534, 5.34, 53.4 and 534 microg L(-1)) of the beta-blocker metoprolol in a flow-through system for seven days. The two lower concentrations represent the environmentally relevant range. Surprisingly, metallothionein mRNA was immediately up-regulated in all treatments. For the two higher concentrations mRNA up-regulation in gills was found for P-glycoprotein after one day, and after four days for pi class glutathione S-transferase, demonstrating elimination and biotransformation processes, respectively. Additionally, catalase and superoxide dismutase were up-regulated in the digestive gland indicating oxidative stress. In all treated mussels a significant up-regulation of heat shock protein mRNA was observed in gills after four days, which suggests protein damage and the requirement for repair processes. Metoprolol was 20-fold bioaccumulated for environmentally relevant concentrations.

Biotransformation and antioxidant enzymes of Limnoperna fortunei detect site impact in watercourses of Córdoba, Argentina.

The golden mussel Limnoperna fortunei was used as a biomonitor of environmental pollution in the Suquía River basin around Córdoba City (Argentina). The sampling sites along the river were chosen according to their increasing levels of pollutants (e.g. heavy metals) as well as biological oxygen demand (BOD) and chemical oxygen demand (COD). A water quality index (WQI) was constructed from the interaction of several normalized factors that affect the aquatic environment, such as the mentioned pollutants and physico-chemical characteristics of the sampling sites. Activity changes of biotransformation enzyme glutathione S-transferase (GST) and the antioxidant enzymes glutathione peroxidase (GPx), glutathione reductase (GR) and catalase (CAT), after exposure to pollutants, served as biomarkers. Membrane bound GST and antioxidant enzymes responded at the most polluted sampling site within 1 day showing increased activities lasting for 4 days. Further sampling was restricted due to no survival of the animals. Antioxidant enzymes GPx, GR and CAT were sensitive responding to the different pollution scenarios, showing good correlation to the chemical characterization.

Biotransformation and antioxidant enzymes of Dreissena polymorpha for detection of site impact in watercourses of Berlin.

This study investigated whether biotransformation and antioxidant enzymes of an organism tolerant to pollution such as the zebra mussel Dreissena polymorpha can be employed to evaluate the extent of urban water pollution. Activity changes of soluble and membrane bound glutathione S-transferase (s- and mGST), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) to environmental stress were explored in precultured mussels exposed for one day to one week at selected sites with reasonable anthropogenic impact. The enzymatic response of whole mussel tissue was compared to gill tissue. Changes in enzyme activity were accordant with the different pollution scenarios, as charges with polychlorinated biphenyls (PCB), dichlorodiphenyltrichloroethane (DDT) and other pesticides, as well as contamination with metals. Whereas in whole mussel tissue all analyzed enzymes responded with elevation, in gills inhibition took place with the exception of mGST. The results confirm the high sensitivity of gills, nevertheless enzymatic changes measured in whole mussel tissue provided the clearest results. Significant changes in GST, CAT and GPx enzyme activity were only observed at water temperatures above 20 degrees C. SOD activity was not restricted by temperatures and serves for this reason as a biomarker for oxidative stress at lower water temperatures. Discrepancies between biological and chemical evaluation of the sampling sites are presented and biomarker responses appraised.

Bioaccumulation of glyphosate and its formulation Roundup Ultra in Lumbriculus variegatus and its effects on biotransformation and antioxidant enzymes.

The bioaccumulation potential of glyphosate and the formulation Roundup Ultra, as well as possible effects on biotransformation and antioxidant enzymes in Lumbriculus variegatus were compared by four days exposure to concentrations between 0.05 and 5 mg L(-1) pure glyphosate and its formulation. Bioaccumulation was determined using (14)C labeled glyphosate. The bioaccumulation factor (BCF) varied between 1.4 and 5.9 for the different concentrations, and was higher than estimated from logP(ow). Glyphosate and its surfactant POEA caused elevation of biotransformation enzyme soluble glutathione S-transferase at non-toxic concentrations. Membrane bound glutathione S-transferase activity was significantly elevated in Roundup Ultra exposed worms, compared to treatment with equal glyphosate concentrations, but did not significantly differ from the control. Antioxidant enzyme superoxide dismutase was significantly increased by glyphosate but in particular by Roundup Ultra exposure indicating oxidative stress. The results show that the formulation Roundup Ultra is of more ecotoxicological relevance than the glyphosate itself.

Multi-xenobiotic-resistance a possible explanation for the insensitivity of bivalves towards cyanobacterial toxins.

Filterfeeders, such as bivalves, are highly affected during toxic cyanobacterial blooms, as they are non-selective and may use the cyanobacteria as main nutrition source. The freshwater mussel Dreissena polymorpha, living in lakes and rivers coexisting with cyanobacteria, was exposed to 100 microg L(-1) microcystin-LR (MC-LR) for up to three days. MC-LR concentration in mussel tissue and surrounding media was quantified by HPLC-PDA during uptake and depuration phase, revealing an immediate, continuous uptake, and release of non-metabolized toxin, and occurrence of reincorporation. The involvement of multi-xenobiotic-resistance protein (P-glycoprotein, P-gp) on the excretion of MC-LR was evidenced by efflux and accumulation version of the Rhodamine Assay as well as on P-gp gene expression. P-gp expression was enhanced after 1 h exposure but no changes were detected after longer (72 h) exposure. P-gp enzyme activity showed a significant increase with exposure time, supporting the hypothesis that P-gp is involved in the excretion of MC-LR. Induction of biotransformation enzyme such as pi-class glutathione S-transferase (piGST) and antioxidant enzyme catalase (CAT) was immediately inhibited and returned to control values only after more than 72 h expose time. Heat shock protein 70 (hsp70) and protein phosphatase 2A (PP2A) gene expression was not changed due to the treatment with cyanobacterial toxin MC-LR.