Highest Test Concentration Research Articles

Toxicity assessment of pyriproxyfen in vertebrate model zebrafish embryos (Danio rerio): A multi biomarker study

Pyriproxyfen (2-[1-methyl-2-(4-phenoxyphenoxy) ethoxy] pyridine) (PPF), a pyridine-based pesticide widely used to control agricultural insect pests and mosquitoes in drinking water sources. However, its ecotoxicological data is limited in aquatic vertebrates particularly in fish. Hence, the present study aimed to evaluate the adverse effect of PPF in zebrafish embryo development (Danio rerio). In order to investigate the impact of PPF, embryos were exposed to 0.16, 0.33 and 1.66 μg/mL (0.52, 1.04 and 5.2 μM, respectively) for 96 hpf and various biomarker indices such as developmental toxicity (edema formation, hyperemia, heart size and scoliosis), oxidative stress (reactive oxygen species (ROS), lipid peroxidation (LPO) and nitric oxide (NO)), antioxidant responses (superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), glutathione peroxidase (GPx) and reduced glutathione (GSH)), biochemical (lactate dehydrogenase (LDH) and acid phosphatase (AP)), neurotoxicity (acetylcholinesterase (AChE)), genotoxicity (apoptosis and DNA damage) and histopathological changes were determined. The results showed that severe developmental deformities and changes in heart rate were observed in embryos treated with highest (1.66 μg/mL) concentration than the control (P < 0.05). Heart size measurement showed that, significant change in heart size (P < 0.01) was observed in embryos of 96 hpf only at 1.66 μg/mL PPF exposure. The oxidative stress was apparent at highest test concentration (1.66 μg/mL) as reflected by the elevated ROS, LPO and NO and changes in antioxidant enzyme activities including SOD, CAT, GST and GPx (P < 0.05). Besides, GSH level and AChE activity were significantly lowered in 1.66 μg/mL PPF exposed group than the control. After 96 hpf of PPF exposure, no significant changes were found in AP activity whereas, a biphasic response was observed in the LDH activity. There was no genotoxic effect in embryos exposed to PPF at 0.16 and 0.33 μg/mL, while significant (P < 0.05) DNA damage and apoptosis were found in 1.66 μg/mL treated group. Histopathological analysis revealed that exposure to PPF at 1.66 μg/mL resulted in thinning of heart muscles, pericardial edema and hyperemia while there was no obvious changes were observed in other treatment groups. Hence, the results of the present study demonstrate that PPF could cause adverse effect on early developmental stages of zebrafish at higher concentration.

Toxicity Evaluation of Acrylamide on the Early Life Stages of the Zebrafish Embryos (&amp;lt;i&amp;gt;Danio rerio&amp;lt;/i&amp;gt;)

Acrylamide is a chemical used mainly in industrial applications and the treatment of drinking and wastewater, making it easy to enter aquatic ecosystems. There are few studies known about the toxicity of acrylamide to aquatic organisms which have shown evidence of a number of histopathological effects. To assess the effects of acrylamide to freshwater fish, Zebrafish (Danio rerio) embryos were exposed to serial concentrations of acrylamide (0, 100, 300, and 500 mg/L) to investigate the acute toxicity effects on teleost embryogenesis. Embryos less than 24 hrs old were exposed under static non-renewal conditions for ten days or until hatching. The toxic endpoints evaluated include: egg/embryo viability, hatchability, and morphological/developmental anomalies during organogenesis. The acute toxicity test resulted in a 48 h-LC50 of 585 mg/L for egg viability. Exposure of embryos significantly reduced hatchability and larval survival, in a concentration dependent manner. Dimethyl sulfoxide (DMSO) was used as a solvent carrier to permeate the uptake of acrylamide through the chorion membrane. No significant damages or complications were observed in embryos exposed to DMSO. At 500 mg/L, the highest test concentration, the survival of embryos was greatly reduced within 24 hrs of exposure. The lower test, 100 mg/L, produced a significant number of developmental anomalies to the Zebrafish that included dorsal tail flexure, severe pericardial edema, facial and cranial defects and decreased heartrate (40 bpm). Premature hatching of embryos and developmental arrest was observed in all concentrations. The severity of these anomalies was concentration-dependent and resulted in low survival rate and high frequency of malformations. These results indicate that acrylamide is teratogenic and provide support for sub-lethal toxicity testing using Zebrafish embryos.

In vitro mutagenic and anti‐mutagenic properties of green synthesised silver nanoparticles

This study is aimed at determining the mutagenic and anti-mutagenic properties of silver nanoparticles (AgNPs) biosynthesised from Streptomyces griseorubens AU2. To the authors’ knowledge, this is the first study about the investigation of these properties for biogenic AgNPs bacterially synthesised. The mutagenic and anti-mutagenic potencies were determined by the Ames Salmonella /microsome mutagenicity test using Salmonella typhimurium TA98 and TA100 strains. After determining the cytotoxic dose of green synthesised AgNPs against S. typhimurium TA98 and TA100 strains, subcytotoxic doses (250, 100 and 50 µg/plate) were used in the assays. Biogenic AgNPs at the tested concentrations exhibited no mutagenic effects in the mutagenicity test conducted with the test strains. Moderate anti-mutagenic effects were observed at high test concentrations. The concentration of 250 µg/plate showed the strongest anti-mutagenic activity on S. typhimurium TA98. The results did not indicate any mutagenic effect against either of the strains used for screening the mutagenicity of the biogenic AgNPs as they were found to be genotoxically safe. It can be concluded that biogenic AgNPs showed great anti-mutagenic attributes, standing as a significant factor with respect to medical, pharmaceutical and cosmetic industries.

Toxicity of a Quinaldine-Based Liquid Organic Hydrogen Carrier (LOHC) System toward Soil Organisms Arthrobacter globiformis and Folsomia candida.

The study aims to establish a preliminary environmental assessment of a quinaldine-based LOHC system composed of hydrogen-lean, partially hydrogenated, and fully hydrogenated forms. We examined their toxicity toward the soil bacteria Arthrobacter globiformis and the Collembola Folsomia candida in two exposure scenarios, with and without soil, to address differences in the bioavailability of the compounds. In both scenarios, no or only slight toxicity toward soil bacteria was observed at the highest test concentration (EC50 > 3397 μmol L-1 and >4892 μmol kg-1 dry weight soil). The effects of the three quinaldines on F. candida in soil were similar, with EC50 values ranging from 2119 to 2559 μmol kg-1 dry weight soil based on nominal concentrations. Additionally, corrected pore-water-concentration-based EC50 values were calculated by equilibrium partitioning using soil/pore-water distribution coefficients. The tests without soil (simulating pore-water exposure) revealed higher toxicity, with LC50 values between 78.3 and 161.6 μmol L-1 and deformation of the protective cuticle. These results assign the compounds to the category "harmful to soil organisms". Potential risks toward the soil environment of the test compounds are discussed on the basis of predicted no-effect concentrations.

Combined Impact of Acute Exposure to Ammonia and Temperature Stress on the Freshwater Mussel Unio pictorum

Nitrogen compounds, particularly ammonia, and temperature are suspected major stressors for aquatic organisms, but little is known about their impact on globally declining freshwater mussels (Unionoida). In this study, we tested the combined effects of ammonia and temperature stress on painter’s mussel (Unio pictorum) survival, filtration behavior, hemocyte abundance, hemocyte mortality and glycogen energy status, at concentrations ranging from 0.3 to 9.0 mg·L−1 total ammonia nitrogen (NH4-N) in 96 h acute exposures at two temperatures, 17 °C and 25 °C and a pH of 8.8. The results indicate a low sensitivity of U. pictorum to elevated ammonia concentrations after short-term exposure, although effects on cell morphology were evident and delayed mortality occurred at the highest test concentration. Most pronounced effects were observed for sublethal physiological endpoints due to elevated temperature, but no synergistic effects with ammonia were evident. Temperature increase resulted in significant effects on tissue glycogen, hyalinocyte mortality and clearance rates. Hemocyte mortalities showed a linear dependency on initial mussel activity as measured by their clearance rate. Since the main stressors tested in this study, ammonia and temperature, are predicted to increase in most freshwater ecosystems, their impact on other freshwater mussel species including different life-stages should be comprehensively assessed.

Assessing multigenerational effects of prednisolone to the freshwater snail, Physa acuta (Gastropoda: Physidae)

Prednisolone (PDS), a potent synthetic glucocorticoid is widely prescribed for its exceptional anti-inflammatory properties. Several studies have detected the environmental presence of PDS in water bodies which has led to an ecological concern for its toxicity to non-target aquatic biota. The present study investigated the effects of exposure to PDS on different life-cycle stages and generations of the freshwater snail, Physa acuta. This continuous exposure over a period of multiple generations resulted in generational impairments at measured endpoints. LOEC values (p<0.001) for PDS exposure ranged from 32 to 4μg/L in exposed F0-F2 generations. Global DNA methylation (% 5-methyl cytosine) of adult progeny was found to be affected at higher test concentrations in comparison to the parent snails. Partially formed to completely missed growth components of shell structure and shell thinning in abnormally underdeveloped PDS exposed snails of F1 and F2 generation, was also observed in this multigenerational exposure experiment. The multigenerational study confirmed P. acuta as a promising bioindicator since critical effects of the long term glucocorticoid exposure opens up the way for further investigations on transgenerational toxicity in environmental toxicology and risk assessment and to monitor glucocorticoid pollution in aqueous ecosystem.

Influence of exogenous chemicals on larval development and survival of the king scallop Pecten maximus (L.)

The commercially important king scallop, Pecten maximus (L.), is the focus of ongoing research to optimise hatchery culture practices. At present, difficulties in the culture of this species are commonly associated with the transition from the larval to juvenile stages, with protracted metamorphosis, low development synchronicity and variable survival. The use of exogenously applied chemical agents, as demonstrated in other bivalve species, has been viewed as a means of resolving these issues. The present study evaluated the effects of the previously untested chemicals KCl, NH4Cl, acetylcholine chloride and GABA, in addition to the previously tested chemical l-DOPA, on the induction of larval metamorphosis and larval survival in P. maximus. A range of concentrations of each chemical, applied over a 48 hour period, were assessed. Larval metamorphic response, based upon the development of functioning gill filaments or secondary shell growth (dissoconch), was low and concentration-dependent. Larval survival was also concentration-dependent, with all chemicals becoming toxic to larvae at high test concentrations. Among the tested chemicals, KCl at 20mM and l-DOPA at 10−6M induced significantly (P<0.05) higher rates of larval metamorphosis, which was improved by 208% and 128% respectively, compared to the controls after 7days. However, whilst the KCl treatment was toxic, reducing survival by 33% compared to the control (P<0.05), the l-DOPA treatment significantly increased survival by 49% compared to the control (P<0.05). Furthermore, the influence of these two chemicals on larval development varied, with KCl promoting dissoconch growth and l-DOPA promoting gill development, suggesting that the pathways influenced by these two chemicals maybe different. In contrast, NH4Cl, acetylcholine chloride and GABA proved ineffective at inducing metamorphosis over the range of concentrations tested. The results of this study provide further evidence supporting the potential use of chemical agents, though further work is required to fully realise the ability to completely synchronise larval metamorphosis in P. maximus for hatchery application. Statement of relevanceInfluence of exogenous chemicals on larval development and survival of the king scallop Pecten maximus (L.)The results of this study provide important advances in our understanding and knowledge of the use of chemical agents in the commercially important bivalve Pecten maximus. This study has identified a new inductive chemical and highlighted considerations for future investigators.

An optimized method to assess ototoxic effects in the lateral line of zebrafish (Danio rerio) embryos

In order to clarify the suitability of the lateral line of zebrafish (Danio rerio) embryos as a model for the screening of ototoxic (neurotoxic) effects, existing neuromast assays were adapted, improved and validated with a series of chemicals known or unknown for their ototoxic potential (caffeine copper sulfate, dichlorvos, 2.4-dinitrotoluene, neomycin, 4-nonylphenol, perfluorooctanesulfonic acid). Present methods were improved by (1) the introduction of a 4-step scoring system, (2) the selection of neuromasts from both the anterior and posterior lateral line systems, (3) a combined DASPEI/DAPI staining applied after both a continuous and pulse exposure scenario, and (4) an additional screening for nuclear fragmentation. Acute toxicities of the model substances were determined by means of the fish embryo test as specified in OECD TG 236, and EC10 concentrations were used as the highest test concentration in the neuromast assay. The enhanced neuromast assay identified known ototoxic substances such as neomycin and copper sulfate as ototoxic at sensitivities similar to those of established methods, with pulse exposure leading to stronger effects than continuous exposure. Except for caffeine, all substances tested (dichlorvos, 2.4-dinitrotoluene, 4-nonylphenol, perfluorooctanesulfonic acid) produced significant toxic effects in neuromasts at EC10 concentrations. Depending on the test substances and their location along the lateral line, specific neuromasts differed in sensitivity. Generally, neuromasts proved more sensitive in the pulse exposure scenario. Whereas for neomycin and copper sulfate neuromasts located along the anterior lateral line were more sensitive, posterior lateral line neuromasts proved more sensitive for the other test substances. Nuclear fragmentation could not only be associated with all test substances, but, albeit at lower frequencies, also with negative controls, and could, therefore, not be assigned specifically to chemical damage. The study thus documented that for a comprehensive evaluation of lateral line damage both neuromasts from the anterior and the posterior lateral line have to be considered. Given the apparently rapid regeneration of hair cells, pulse exposure seems more appropriate for the identification of lateral line neurotoxicity than continuous exposure.

Population and life‐stage–specific effects of two herbicide formulations on the aquatic development of European common frogs (Rana temporaria)

Environmental contamination is suggested to contribute to amphibian population declines. However, the effects of a contaminant on a particular amphibian species can differ among populations. The authors investigated the toxic effects of 2 herbicide formulations on different populations and on representative developmental stages of the European common frog (Rana temporaria). Larvae from forest populations were more sensitive to a commonly used glyphosate-based herbicide compared with individuals from agrarian land. Median lethal concentrations correlated with measured glyphosate levels in the breeding ponds, which may be a sign of evolved tolerances. The reverse result was observed for a less commonly used cycloxydim-based herbicide. Effects of the glyphosate-based herbicide were stronger for earlier larval stages compared with later larval stages. Hence, applications in early spring (when early larvae are present in breeding ponds) pose greater risk concerning acute toxic effects on R. temporaria. With regard to late larval stages, short exposure (96 h) of prometamorphic larvae prolonged time to metamorphosis, but only at the highest test concentration that did not significantly induce mortality. This could be due to impairment of the thyroid axis. Notably, nearly all test concentrations of the 2 herbicides provoked growth retardation. Further research on how evolved or induced tolerances are acquired, actual contamination levels of amphibian habitats, and potential endocrine effects of glyphosate-based herbicides is necessary. Environ Toxicol Chem 2017;36:190-200. © 2016 SETAC.

Effects of a Standardized Phenolic-Enriched Maple Syrup Extract on β-Amyloid Aggregation, Neuroinflammation in Microglial and Neuronal Cells, and β-Amyloid Induced Neurotoxicity in Caenorhabditis elegans.

Published data supports the neuroprotective effects of several phenolic-containing natural products, including certain fruit, berries, spices, nuts, green tea, and olive oil. However, limited data are available for phenolic-containing plant-derived natural sweeteners including maple syrup. Herein, we investigated the neuroprotective effects of a chemically standardized phenolic-enriched maple syrup extract (MSX) using a combination of biophysical, in vitro, and in vivo studies. Based on biophysical data (Thioflavin T assay, transmission electron microscopy, circular dichroism, dynamic light scattering, and zeta potential), MSX reduced amyloid β1-42 peptide (Aβ1-42) fibrillation in a concentration-dependent manner (50-500μg/mL) with similar effects as the neuroprotective polyphenol, resveratrol, at its highest test concentration (63.5 % at 500μg/mL vs. 77.3 % at 50μg/mL, respectively). MSX (100μg/mL) decreased H2O2-induced oxidative stress (16.1 % decrease in ROS levels compared to control), and down-regulated the production of lipopolysaccharide (LPS)-stimulated inflammatory markers (22.1, 19.9, 74.8, and 87.6 % decrease in NOS, IL-6, PGE2, and TNFα levels, respectively, compared to control) in murine BV-2 microglial cells. Moreover, in a non-contact co-culture cell model, differentiated human SH-SY5Y neuronal cells were exposed to conditioned media from BV-2 cells treated with MSX (100μg/mL) and LPS or LPS alone. MSX-BV-2 media increased SH-SY5Y cell viability by 13.8 % compared to media collected from LPS-BV-2 treated cells. Also, MSX (10μg/mL) showed protective effects against Aβ1-42 induced neurotoxicity and paralysis in Caenorhabditis elegans in vivo. These data support the potential neuroprotective effects of MSX warranting further studies on this natural product.

Comparative sensitivity of juvenile and adult Potamopyrgus antipodarum (Mollusca: Hydrobiidae) under chronic exposure to cadmium and tributyltin

ABSTRACTTo investigate a potential extension of a partial life cycle test protocol to a full life cycle test design, a comparative sensitivity analysis with juvenile and adult Potamopyrgus antipodarum was performed. Neonates and adult snails were exposed to the metal cadmium (Cd) and the endocrine disruptor tributyltin (TBT) at nominal concentrations ranging from 1.56 to 50 μg Cd/L and from 25 to 1,000 ng TBT-Sn/L. The experiments were performed over 28 days at 16°C in a semi-static test design. Mortality was assessed for both life stages. Juvenile snails' specific growth rate and reproduction of adults were investigated as main endpoints. We determined effects on snails' survival, juvenile growth and embryo numbers in the brood pouch of adult snails under exposure to both chemicals. Juvenile control mortality was between 25% and 30% and significantly higher than in the control groups with adult snails. A higher sensitivity of juvenile snails compared to adults was observed for the endpoint mortality. Calculated LC50 in Cd exposed snails was 38.2 μg/L for adults and 15.0 μg/L for juvenile snails. Significant effects on mortality in TBT exposed adult snails occurred at the highest test concentration only with a LC50 of 535 ng Sn/L. Juvenile survival was significantly affected at 50.8 ng Sn/L and higher concentrations. Effect concentrations for the main endpoints reproduction and juvenile growth show comparable sensitivities. For Cd exposed groups, EC50 values were 11.3 μg/L for the endpoint reproduction in adult snails and 3.82 μg/L for juvenile growth with overlapping confidence intervals. TBT also significantly affected juvenile snails' growth (EC50: 178 ng Sn /L). EC50 for embryo numbers was 125 ng TBT-Sn/L. Results indicate the manageability of a FLC test starting with newly hatched snails. Precautions have to be taken to guarantee a sufficient number of surviving snails until adulthood so that reproduction can be assessed. For final decision for the practicability of a FLC, further tests are needed.

Current limitations of biodegradation screening tests and prediction of biodegradability: A focus on fragrance substances

Abstract Key parameters of OECD biodegradation screening tests were analyzed for impact on the assessment of biodegradation potential of selected fragrance compounds. Comparison of official Japanese OECD 301C test results for 48 predominantly readily biodegradable chemicals with recent OECD 301F screening data demonstrated a significantly higher occurrence of false negatives potentially attributable to inoculum limitations for the former, while for the latter, restriction to 28 days was frequently limiting. Influence of test concentration was studied for two groups of quaternary carbon-containing compounds under high (OECD 301F) and low (OECD 301D) concentrations and extended incubations. Ionones were generally ultimately biodegradable within 28 days or readily biodegradable, whereas damascones reached ultimate biodegradation in OECD 301D tests only and required extended incubations. Predictions from Biowin and Catalogic models were analyzed for ionones, damascones and other selected structural groups of fragrance compounds. Comparison with previously published experimental data revealed significant differences in results between OECD screening tests, thus illustrating the shortcomings of currently available data collections and the potential risk of training prediction models with false negative results, particularly if used for models that are primarily based on data from one specific test. In addition, the analysis revealed specific limitations for prediction models that cannot take into account positioning of fragments within a structure. As a result, it appears that the presence of more than one unfavorable fragment usually does not allow reaching currently accepted threshold scores that would identify a given compound as biodegradable. The outcome of the analyses of this study underlines the need to take into account results from OECD screening tests that are best adapted to physico-chemical properties of fragrances (hydrophobicity, volatility and inhibition potential at high test concentration) and conducted at reduced stringency compared to OECD guidelines (extended incubation owing to reduced bioavailability). Negative MITI database results should be critically reviewed, particularly for quaternary carbon-containing compounds. For future efforts aimed at designing more environmentally benign chemicals, the apparent underestimations will have to be overcome by providing more realistic experimental data that should subsequently be used to improve current biodegradation prediction models for better guidance.

Ethylhexylglycerin: a low-risk, but highly relevant, sensitizer in 'hypo-allergenic' cosmetics.

Ethylhexylglycerin is a relatively new cosmetic ingredient that is used for its surfactant, emollient, skin-conditioning and antimicrobial properties. Since 2002, it has been occasionally reported as a contact allergen. To report on 13 patients who presented with allergic contact dermatitis caused by ethylhexylglycerin, evaluated at two Belgian university patch test clinics during the period 1990-2015. The patients were patch tested with the European baseline series, a cosmetic series, and - if indicated - additional series. Both the cosmetic products used and their single ingredients were patch tested. All but one of the ethylhexylglycerin-allergic patients were female, with a median age of 43 years (range: 29-81 years), most often suffering from dermatitis on the face, and sometimes on the hands and/or axillae. As the culprit products, leave-on cosmetics were identified, including a high number of proclaimed 'hypo-allergenic' and 'preservative-free' facial creams, sun protection creams, and deodorants. Ethylhexylglycerin is a rare, but highly relevant, cosmetic sensitizer, even in those products advertised to be safe for consumers. Targeted patch testing with ethylhexylglycerin 5% pet. is very useful, and routine patch testing in a cosmetic series may be considered. Higher test concentrations might be indicated in selected cases.

The effects of plant essential oils on escape response and mortality rate of Aedes aegypti and Anopheles minimus.

The High Throughput Screening System (HITSS) has been applied in insecticide behavioral response studies with various mosquito species. In general, chemical or natural compounds can produce a range of insect responses: contact irritancy, spatial repellency, knock-down, and toxicity. This study characterized these actions in essential oils derived from citronella, hairy basil, catnip, and vetiver in comparison to DEET and picaridin against Aedes aegypti and Anopheles minimus mosquito populations. Results indicated the two mosquito species exhibited significantly different (P<0.05) contact irritant escape responses between treatment and control for all tested compound concentrations, except with the minimum dose of picaridin (P>0.05) against Ae. aegypti. Spatial repellency responses were elicited in both mosquito species when exposed to all compounds, but the strength of the repellent response was dependent on compound and concentration. Data show that higher test concentrations had greatest toxic effects on both mosquito populations, but vetiver had no toxic effect on Ae. aegypti and picaridin did not elicit toxicity in either Ae. aegypti or An. minimus at any test concentration. Ultimately, this study demonstrates the ability of the HITSS assay to guide selection of effective plant essential oils for repelling, irritating, and killing mosquitoes.

Acute Toxicity and Environmental Risks of Five Veterinary Pharmaceuticals for Aquatic Macroinvertebrates.

Due to the high use of antibiotics and antiparasitics for the treatment of livestock, there is concern about the potential impacts of the release of these compounds into freshwater ecosystems. In this context, the present study quantified the acute toxicity of two antibiotics (sulfadiazine and sulfadimidine), and three antiparasitic agents (flubendazole, fenbendazole, ivermectin) for nine freshwater invertebrate species. These experiments revealed a low degree of toxicity for the sulfonamide antibiotics, with limited implications in the survival of all test species at the highest test concentrations (50 and 100 mg/L). In contrast, all three antiparasitic agents indicated on the basis of their acute toxicity risks for the aquatic environment. Moreover, chronic toxicity data from the literature for antiparasitics, including effects on reproduction in daphnids, support the concern about the integrity of aquatic ecosystems posed by releases of these compounds. Thus, these pharmaceuticals warrant further careful consideration by environmental risk managers.

Short-term fish reproduction assays with methyl tertiary butyl ether with zebrafish and fathead minnow: Implications for evaluation of potential for endocrine activity.

The authors report on short-term fish reproduction assays in zebrafish and fathead minnow conducted to examine the potential for methyl tertiary butyl ether (MTBE) to cause effects on the endocrine system. Both studies were performed under good laboratory practice and in accordance with Organisation for Economic Co-operation and Development and US Environmental Protection Agency test guidelines. The results of the first study demonstrated that exposure to a high test concentration (147 mg/L) of MTBE impaired reproductive output of female zebrafish, evident by a reduction in fecundity. Based on the endpoints evaluated in the present study however, there was no supporting evidence to indicate that this effect was caused by disruption of or interaction with the endocrine system. In the second study, fathead minnows exposed to a wider but lower range of test concentrations showed no effects on any reproductive parameter of male or female fish, at the maximum recommended testing concentration of 100 mg/L (62 mg/L measured). The results of these 2 guideline studies indicate that MTBE does not interact with the hypothalamic-pituitary-gonadal axis of zebrafish or fathead minnow.

Effect of Chlorpyrifos Ethyl on Acetylcholinesterase Activity in Climbing Perch (Anabas testudineus, Bloch, 1972).

The high use of pesticides in intensive rice farming in the Mekong Delta constitutes a potential hazard to the environment and to people's health. Chlorpyrifos ethyl (CPF) is a commonly used organophosphate (OP) insecticide, but information about its potential negative impacts on the aquatic environment in the Mekong Delta is scarce. Both acute and subacute toxicity tests were performed in a static nonrenewable system to investigate the effects of CPF on brain acetylcholinesterase (AChE) activity in native climbing perch fingerlings (Anabas testudineus, Bloch, 1972). Environmental parameters, such as dissolved oxygen, water temperature, and pH, were similar to field conditions in the Mekong Delta. In a 96-h lethal concentration (LC50) test, fingerlings of climbing perch were randomly exposed to five levels of CPF ranging from 0.8 to 4.5ppm. Five sublethal levels of CPF (1, 5, 10, 15, and 20% of the 96-h LC50 value) were tested to assess the sensitivity and recovery of the brain AChE activity in climbing perch fingerlings exposed to CPF. The results showed that CPF were moderately toxic to climbing perch with a 96-h median LC50 of 1.73ppm. CPF also caused long-term AChE inhibition with 70% inhibition remaining after 96h for the four highest test concentrations. The recovery of brain AChE activity in fish placed in CPF-free water was very slow, and after 7days the brain AChE activity was still significant lower in fish from the four highest concentrations compared with the control. The results from this study indicate that OP insecticides, such as CPF, can have long-lasting sublethal effects on aquatic species in the Mekong Delta.

Effects of Louisiana crude oil on the sheepshead minnow (Cyprinodon variegatus) during a life-cycle exposure to laboratory oiled sediment.

Determining the long-term effects of crude oil exposure is critical for ascertaining population-level ecological risks of spill events. A 19-week complete life-cycle experiment was conducted with the estuarine sheepshead minnow (Cyprinodon variegatus) exposed to reference (uncontaminated) sediment spiked with laboratory weathered South Louisiana crude (SLC) oil at five concentrations as well as one unspiked sediment control and one seawater (no sediment) control. Newly hatched larvae were exposed to the oiled sediments at measured concentrations of < 1 (sediment control), 50, 103, 193, 347, and 711 mg total polyaromatic hydrocarbons (tPAH)/kg dry sediment. Juveniles were exposed through the reproductively active adult phase at measured concentrations of <1 (sediment control), 52, 109, 199, 358, and 751 mg tPAH/kg sediment. Throughout the exposure, fish were assessed for growth, survival, and reproduction. Resulting F1 embryos were then collected, incubated, and hatched in clean water to determine if parental full life-cycle exposure to oiled sediment produced trans-generational effects. Larvae experienced significantly reduced standard length (5-13% reduction) and wet weight (13-35% reduction) at concentrations at and above 50 and 103 mg tPAH/kg sediment, respectively. At 92 and 132 days post hatch (dph), standard length was reduced (7-13% reduction) at 199 and 109 mg tPAH/kg dry sediment, respectively, and wet weight for both time periods was reduced at concentrations at and above 109 mg tPAH/kg dry sediment (21-38% reduction). A significant reduction (51-65%) in F0 fecundity occurred at the two highest test concentrations, but no difference was observed in F1 embryo survival. This study is the first to report the effects of chronic laboratory exposure to oiled sediment, and will assist the development of population models for evaluating risk to benthic spawning fish species exposed to oiled sediments. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1627-1639, 2016.

Acetylcholinesterase in zebrafish embryos as a tool to identify neurotoxic effects in sediments.

In order to clarify the suitability of zebrafish (Danio rerio) embryos for the detection of neurotoxic compounds, the acetylcholinesterase assay was adapted and validated with a series of priority pollutants listed as relevant for the European water policy (Aroclor 1254, 2,3-benzofuran, bisphenol A, chlorpyrifos, paraoxon-methyl, quinoline, and methyl mercury chloride) as well as acetonic extracts from three sediments of known contamination. The acute toxicities of the model substances and the sediment extracts were determined by means of the fish embryo test as specified in OECD TG 236, and concentrations as low as the effective concentration at 10% inhibition (EC10) were used as the highest test concentration in the acetylcholinesterase test in order to avoid nonspecific systemic effects mimicking neurotoxicity. Among the model compounds, only the known acetylcholinesterase inhibitors paraoxon-methyl and chlorpyrifos produced a strong inhibition to about 20 and 33%, respectively, of the negative controls. For the sediment extracts, a reduction of acetylcholinesterase activity to about 60% could only be shown for the Vering Canal sediment extracts; this could be correlated to high contents of acetylcholinesterase-inhibiting polycyclic aromatic hydrocarbons (PAHs) as identified by chemical analyses. Co-incubation of the Vering Canal sediment extracts with chlorpyrifos at EC10 concentrations each did not significantly increase the inhibitory effect of chlorpyrifos, indicating that the mode of action of acetylcholinesterase inhibition by the sediment-borne PAHs is different to that of the typical acetylcholinesterase blocker chlorpyrifos. Overall, the study documents that zebrafish embryos represent a suitable model not only to reveal acetylcholinesterase inhibition, but also to investigate various modes of neurotoxic action.

Increased RO concentrate toxicity following application of antiscalants – Acute toxicity tests with the amphipods Gammarus pulex and Gammarus roeseli

In reverse osmosis, a frequently used technology in water desalination processes, wastewater (RO concentrate) is generated containing the retained solutes as well as so-called antiscalants (AS), i.e. chemical substances that are commonly applied to prevent membrane-blocking. In this study, a risk assessment of a possible discharge of concentrate into a small stream was conducted. The acute toxicity of two concentrates containing two different ASs and of concentrate without AS to the amphipods Gammarus pulex and Gammarus roeseli was studied. Mortality of gammarids exposed to the concentrate without AS was not different to the control, whereas concentrates including ASs caused mortality rates up to 100% at the highest test concentrations after 168 h. Resulting EC50-values were 36.2–39.4% (v/v) after 96 h and 26.6–58.0% (v/v) after 168 h. These results suggest that the ecotoxicological relevance of antiscalants is greater than currently assumed.