Showing papers in "Ecotoxicology in 2012"

Neonicotinoids in bees: a review on concentrations, side-effects and risk assessment

Abstract: Neonicotinoid insecticides are successfully applied to control pests in a variety of agricultural crops; however, they may not only affect pest insects but also non-target organisms such as pollinators. This review summarizes, for the first time, 15 years of research on the hazards of neonicotinoids to bees including honey bees, bumble bees and solitary bees. The focus of the paper is on three different key aspects determining the risks of neonicotinoid field concentrations for bee populations: (1) the environmental neonicotinoid residue levels in plants, bees and bee products in relation to pesticide application, (2) the reported side-effects with special attention for sublethal effects, and (3) the usefulness for the evaluation of neonicotinoids of an already existing risk assessment scheme for systemic compounds. Although environmental residue levels of neonicotinoids were found to be lower than acute/chronic toxicity levels, there is still a lack of reliable data as most analyses were conducted near the detection limit and for only few crops. Many laboratory studies described lethal and sublethal effects of neonicotinoids on the foraging behavior, and learning and memory abilities of bees, while no effects were observed in field studies at field-realistic dosages. The proposed risk assessment scheme for systemic compounds was shown to be applicable to assess the risk for side-effects of neonicotinoids as it considers the effect on different life stages and different levels of biological organization (organism versus colony). Future research studies should be conducted with field-realistic concentrations, relevant exposure and evaluation durations. Molecular markers may be used to improve risk assessment by a better understanding of the mode of action (interaction with receptors) of neonicotinoids in bees leading to the identification of environmentally safer compounds.

Practical considerations for conducting ecotoxicity test methods with manufactured nanomaterials: what have we learnt so far?

Abstract: This review paper reports the consensus of a technical workshop hosted by the European network, NanoImpactNet (NIN). The workshop aimed to review the collective experience of working at the bench with manufactured nanomaterials (MNMs), and to recommend modifications to existing experimental methods and OECD protocols. Current procedures for cleaning glassware are appropriate for most MNMs, although interference with electrodes may occur. Maintaining exposure is more difficult with MNMs compared to conventional chemicals. A metal salt control is recommended for experiments with metallic MNMs that may release free metal ions. Dispersing agents should be avoided, but if they must be used, then natural or synthetic dispersing agents are possible, and dispersion controls essential. Time constraints and technology gaps indicate that full characterisation of test media during ecotoxicity tests is currently not practical. Details of electron microscopy, dark-field microscopy, a range of spectroscopic methods (EDX, XRD, XANES, EXAFS), light scattering techniques (DLS, SLS) and chromatography are discussed. The development of user-friendly software to predict particle behaviour in test media according to DLVO theory is in progress, and simple optical methods are available to estimate the settling behaviour of suspensions during experiments. However, for soil matrices such simple approaches may not be applicable. Alternatively, a Critical Body Residue approach may be taken in which body concentrations in organisms are related to effects, and toxicity thresholds derived. For microbial assays, the cell wall is a formidable barrier to MNMs and end points that rely on the test substance penetrating the cell may be insensitive. Instead assays based on the cell envelope should be developed for MNMs. In algal growth tests, the abiotic factors that promote particle aggregation in the media (e.g. ionic strength) are also important in providing nutrients, and manipulation of the media to control the dispersion may also inhibit growth. Controls to quantify shading effects, and precise details of lighting regimes, shaking or mixing should be reported in algal tests. Photosynthesis may be more sensitive than traditional growth end points for algae and plants. Tests with invertebrates should consider non-chemical toxicity from particle adherence to the organisms. The use of semi-static exposure methods with fish can reduce the logistical issues of waste water disposal and facilitate aspects of animal husbandry relevant to MMNs. There are concerns that the existing bioaccumulation tests are conceptually flawed for MNMs and that new test(s) are required. In vitro testing strategies, as exemplified by genotoxicity assays, can be modified for MNMs, but the risk of false negatives in some assays is highlighted. In conclusion, most protocols will require some modifications and recommendations are made to aid the researcher at the bench.

Effects of imidacloprid, a neonicotinoid pesticide, on reproduction in worker bumble bees ( Bombus terrestris )

Abstract: Bumble bees are important pollinators whose populations have declined over recent years, raising widespread concern. One conspicuous threat to bumble bees is their unintended exposure to trace residues of systemic neonicotinoid pesticides, such as imidacloprid, which are ingested when bees forage on the nectar and pollen of treated crops. However, the demographic consequences for bumble bees of exposure to dietary neonicotinoids have yet to be fully established. To determine whether environmentally realistic levels of imidacloprid are capable of making a demographic impact on bumble bees, we exposed queenless microcolonies of worker bumble bees, Bombus terrestris, to a range of dosages of dietary imidacloprid between zero and 125 μg L(-1) and examined the effects on ovary development and fecundity. Microcolonies showed a dose-dependent decline in fecundity, with environmentally realistic dosages in the range of 1 μg L(-1) capable of reducing brood production by one third. In contrast, ovary development was unimpaired by dietary imidacloprid except at the highest dosage. Imidacloprid reduced feeding on both syrup and pollen but, after controlling statistically for dosage, microcolonies that consumed more syrup and pollen produced more brood. We therefore speculate that the detrimental effects of imidacloprid on fecundity emerge principally from nutrient limitation imposed by the failure of individuals to feed. Our findings raise concern about the impact of neonicotinoids on wild bumble bee populations. However, we recognize that to fully evaluate impacts on wild colonies it will be necessary to establish the effect of dietary neonicotinoids on the fecundity of bumble bee queens.

Lethal effect of imidacloprid on the coccinellid predator Serangium japonicum and sublethal effects on predator voracity and on functional response to the whitefly Bemisia tabaci

Abstract: Neonicotinoid insecticides are widely used for controlling sucking pests, and sublethal effects can be expected in beneficial arthropods like natural enemies. Serangium japonicum is an important predator in many agricultural systems in China, and a potential biological control agent against Bemisia tabaci. We evaluated the toxicity of imidacloprid to S. japonicum and its impact on the functional response to B. tabaci eggs. S. japonicum adults exposed through contact to dried residues of imidacloprid at the recommended field rate on cotton against B. tabaci (4 g active ingredient per 100 l, i.e. 40 ppm [part per million]), and reduced rates (25, 20, 15 and 10 ppm) for 24 h showed high mortality rates. The mortality induced by a lowest rate, 5 ppm, was not significantly different than the control group and thus it was considered as a sublethal rate. The lethal rate 50 and hazard quotient (HQ) were estimated to be 11.54 ppm and 3.47 respectively, indicating a risk for S. japonicum in treated fields (HQ > 2). When exposed to dried residues of imidacloprid at the sublethal rate (5 ppm) on cotton leaves, functional response of S. japonicum to B. tabaci eggs was affected with an increase in handling time and a reduction in peak consumption of eggs. Imidacloprid residues also disturbed predator voracity, the number of B. tabaci eggs consumed on treated leaves being significantly lower than on untreated leaves. All effects disappeared within a few hours after transfer to untreated cotton leaves. Imidacloprid systemically applied at the recommended field rate (for cotton) showed no toxicity to S. japonicum, nor affected the functional response of the predator. Sublethal effects of imidacloprid on S. japonicum observed in our study likely negatively affect S. japonicum development and reproductive capacity and may ultimately reduce predator population growth. These results hint at the importance of assessing potential effects of imidacloprid on S. japonicum for developing effective integrated pest management programs of B. tabaci in China.

DNA damage and oxidative stress induced by endosulfan exposure in zebrafish (Danio rerio).

Abstract: Endosulfan (6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzo-dioxathiepin-3-oxide), an organochlorine pesticide, is prevalently used all around the world. It is considered to be a new candidate for the persistent organic pollutants group. Endosulfan residues in the environment may cause serious damage to ecosystems, especially in aquatic environments. The present study was conducted to investigate the effect of endosulfan on antioxidant enzymes [catalase (CAT) and superoxide dismutase (SOD)], reactive oxygen species (ROS) generation and DNA damage in zebrafish. Male and female zebrafish were separated and exposed to a control solution and four concentrations of endosulfan (0.01, 0.1, 1, and 10 μg L−1) and were sampled after 7, 14, 21, and 28 days. It is noteworthy that the present research explored the correlation among the three indicators induced by endosulfan. Low endosulfan concentrations (0.01 μg L−1) induced a slight increase of SOD and CAT activity, which kept ROS in a stable level. High endosulfan concentration (10 μg L−1) induced excessive ROS production which exceeded the capacity of the cellular antioxidants and exhausted the enzyme including CAT and SOD. The DNA damage of zebrafish was evaluated by single-cell gel electrophoresis and was enhanced with increasing endosulfan concentration. In conclusion, the present study showed that endosulfan (0.01–10 μg L−1) has toxic effects on zebrafish.

Risk assessment of chlortetracycline, oxytetracycline, sulfamethazine, sulfathiazole, and erythromycin in aquatic environment: are the current environmental concentrations safe?

Abstract: To understand potential risks of major pharmaceutical residues in waters, we evaluated ecotoxicities of five major veterinary pharmaceuticals, i.e., chlortetracycline, oxytetracycline, sulfamethazine, sulfathiazole, and erythromycin, which have been frequently detected in freshwater environment worldwide. We conducted acute and chronic toxicity tests using two freshwater invertebrates (Daphnia magna and Moina macrocopa) and a fish (Oryzias latipes). In general, D. magna exhibited greater sensitivity than M. macrocopa, and chronic reproduction was the most sensitive endpoints for both organisms. The population growth rate was adversely influenced by exposure to chlortetracycline, sulfamethazine, or sulfathiazole in water fleas, but reduction in population size was not expected. In O. latipes, the tested pharmaceuticals affected several reproduction related endpoints including time to hatch and growth. Based on the toxicity values from the present study and literature, algae appeared to be the most sensitive organism, followed by Daphnia and fish. Hazard quotients derived from measured environmental concentrations (MECs) and predicted no effect concentrations (PNECs) for erythromycin and oxytetracycline exceeded unity, suggesting that potential ecological effects at highly contaminated sites cannot be ruled out. Long-term consequences of veterinary pharmaceutical contamination in the environment deserve further investigation.

Ecotoxicological effects of ciprofloxacin on freshwater species: data integration and derivation of toxicity thresholds for risk assessment

Abstract: Although antibiotics have been increasingly used and detected in natural samples, their ecotoxicological effects on aquatic wildlife are not yet extensively studied. Considering the environmental threat posed by the biological activity of antibiotics it is quite relevant to assess the resulting impact, especially on sub-lethal endpoints. As such, this study evaluated the effects of ciprofloxacin on Pseudokirchneriella subcapitata and Lemna minor growth, on the survival and reproduction of Daphnia magna and on Gambusia holbrooki survival. The risks associated with ciprofloxacin effects on non-target organisms were quantified through the calculation of the PEC/PNEC ratio. Overall, the toxicity values obtained (at the mg L−1 level) were higher than the environmental concentrations. P. subcapitata and L. minor were more sensitive under short-term exposures than D. magna and G. holbrooki. No acute toxicity was observed for fish. The chronic assay with D. magna evidenced that long term exposures to lower concentrations of this antibiotic induced impairments on its life-history parameters. Such outcome may pre-empt potential damages on the long-term maintenance of natural populations continuously exposed to the input of antibiotics. Indeed, the PEC/PNEC ratios showed that ciprofloxacin represents a risk for the most sensitive aquatic organisms, since the defined threshold of an acceptable risk was considerably surpassed.

Assessment of physiological sublethal effects of imidacloprid on the mirid bug Apolygus lucorum (Meyer-Dür).

Abstract: Apolygus lucorum (Meyer-Dur) (Hemiptera: Miridae) is currently one of major mirid pests in the Yangtze River and the Yellow River regions in China. Imidacloprid (neonicotinoid) is widely used against pierce-sucking pest insects, including against A. lucorum. In addition to its direct lethal effect, multiple negative sublethal effects may also occur in exposed insects. We assessed potential sublethal effects of imidacloprid on some biological characteristics of A. lucorum with the aim of increasing rational use of imidacloprid against that cotton pest. The lethal toxicity of imidacloprid on adults of A. lucorum was determined in laboratory conditions by a topical application bioassay (LD50 = 6.70 ng a.i. [active ingredient]/A. lucorum adult). We also estimated a sublethal dose, LD5 (0.38 ng a.i./adult), a low lethal dose, LD25 (1.96 ng a.i./adult), and moderate lethal dose, LD40 (3.97 ng a.i./adult). The sublethal dose of imidacloprid (LD5) shortened the pre-oviposition period of females but increased the time required for eggs to develop (i.e. longer embryogenesis). The low lethal dose (LD25) also reduced the pre-oviposition period. Females exposed to the LD40 laid eggs that developed faster but overall percentage of eggs hatching was reduced. LD25 and LD40 reduced longevity of males but not of females. In addition, the susceptibility to seven insecticides generally used on Chinese crops was not modified in A. lucorum previously exposed to the LD25 of imidacloprid. Our results demonstrate sublethal effects of low doses of imidacloprid on A. lucorum (notably on pre-oviposition period and egg development) which may have an impact on population dynamics of this pest.

Short-term and transgenerational effects of the neonicotinoid nitenpyram on susceptibility to insecticides in two whitefly species

Abstract: The cosmopolitan silverleaf whitefly, Bemisia tabaci which had coexisted with Trialeurodes vaporariorum in Northern China for many years, has become the dominant species in the last years. Recent reports show that it is gradually displacing the other greenhouse whitefly species. Neonicotinoid, which includes nitenpyram, is a major group of insecticides used against whiteflies in various crops. When exposed to low doses of insecticides, insects may develop resistance by adapting physiologically. The short- and long-term effects of nitenpyram on insecticide sensitivity in B. tabaci biotype B and T. vaporariorum adult populations have been compared in the present study. After being exposed to LC25 of nitenpyram for 24 h, the B. tabaci biotype B adults showed no significant change in susceptibility to nitenpyram or to five other insecticides: imidacloprid, acetamiprid, abamectin, chlorpyrifos and beta-cypermethrin. By contrast, exposure to the LC25 of nitenpyram for 24 h led to a significant increase in the susceptibility of T. vaporariorum to nitenpyram and imidacloprid, by 1.8- and 2-fold, respectively. When exposed for seven generations to the LC25 of nitenpyram, B. tabaci developed 6-fold resistance to nitenpyram, and 3.1- and 5-fold cross-resistance to imidacloprid and acetamiprid, respectively, whereas T. vaporariorum developed lower resistance (3.7-fold) to the nitenpyram and very low cross-resistance to imidacloprid (2.5-fold). The higher adaptable nature of B. tabaci (demonstrated here in the case of nitenpyram) when exposed to low doses of insecticides may provide a selective advantage when competing with T. vaporariorum in crops.

Olive oil mill wastewaters before and after treatment: a critical review from the ecotoxicological point of view.

Abstract: The olive oil mill wastewater (OMW) is a problematic and polluting effluent which may degrade the soil and water quality, with critical negative impacts on ecosystems functions and services provided The main purpose of this review paper is presenting the state of the art of OMW treatments focusing on their efficiency to reduce OMW toxicity, and emphasizing the role of ecotoxicological tests on the evaluation of such efficiency before the up-scale of treatment methodologies being considered In the majority of research works, the reduction of OMW toxicity is related to the degradation of phenolic compounds (considered as the main responsible for the toxic effects of OMW on seed germination, on bacteria, and on different species of soil and aquatic invertebrates) or the decrease of chemical oxygen demand content, which is not scientifically sound Batteries of ecotoxicological tests are not applied before and after OMW treatments as they should be, thus leading to knowledge gaps in terms of accurate and real assessment of OMW toxicity Although the toxicity of OMW is usually high, the evaluation of effects on sub-lethal endpoints, on individual and multispecies test systems, are currently lacking, and the real impacts yielded by its dilution, in freshwater trophic chains of receiving systems can not be assessed As far as the terrestrial compartment is considered, ecotoxicological data available include tests only with plants and the evaluation of soil microbial parameters, reflecting concerns with the impacts on crops when using OMW for irrigation purposes The evaluation of its ecotoxicity to other edaphic species were not performed giving rise to a completely lack of knowledge about the consequences of such practice on other soil functions OMW production is a great environmental problem in Mediterranean countries; hence, engineers, chemists and ecotoxicologists should face this problem together to find an ecologically friend solution

Assessment of microcystin distribution and biomagnification in tissues of aquatic food web compartments from a shallow lake and evaluation of potential risks to public health

Abstract: The objectives of this study were: (1) to examine the distribution and bioaccumulation of microcystins in the main components of the food web (phytoplankton, zooplankton, crayfish, shrimp, mussel, snail, fish, frog) of Lake Pamvotis (NW Greece), (2) to investigate the possibility of microcystin biomagnification and (3) to evaluate the potential threat of the contaminated aquatic organisms to human health. Significant microcystin concentrations were detected in all the aquatic organisms during two different periods, with the higher concentrations observed in phytoplankton and the lower in fish species and frogs. This is the first study reporting microcystin accumulation in the body of the freshwater shrimp Atyaephyra desmsaresti, in the brain of the fish species common carp (Cyprinus carpio) and in the skin of the frog Rana epirotica. Although there was no evidence for microcystin biomagnification, the fact that microcystins were found in lake water and in the tissues of aquatic organisms, suggests that serious risks to animal and public health are possible to occur. In addition, it is likely to be unsafe to consume aquatic species harvested in Lake Pamvotis due to the high-concentrations of accumulated microcystins.

Differential genotoxicity of Roundup ® formulation and its constituents in blood cells of fish ( Anguilla anguilla ): considerations on chemical interactions and DNA damaging mechanisms

Abstract: It has been widely recognized that pesticides represent a potential threat in aquatic ecosystems. However, the knowledge on the genotoxicity of pesticides to fish is still limited. Moreover, genotoxic studies have been almost exclusively focused on the active ingredients, whereas the effect of adjuvants is frequently ignored. Hence, the present study addressed the herbicide Roundup®, evaluating the relative contribution of the active ingredient (glyphosate) and the surfactant (polyethoxylated amine; POEA) to the genotoxicity of the commercial formulation on Anguilla anguilla. Fish were exposed to equivalent concentrations of Roundup® (58, 116 μg L−1), glyphosate (17.9, 35.7 μg L−1) and POEA (9.3, 18.6 μg L−1), during 1 and 3 days. The comet assay was applied to blood cells, either as the standard procedure, or with an extra step involving DNA lesion-specific repair enzymes in an attempt to clarify DNA damaging mechanisms. The results confirmed the genotoxicity of Roundup®, also demonstrating the genotoxic potential of glyphosate and POEA individually. Though both components contributed to the overall genotoxicity of the pesticide formulation, the sum of their individual effects was never observed, pointing out an antagonistic interaction. Although POEA is far from being considered biologically inert, it did not increase the risk associated to glyphosate when the two were combined. The analysis of oxidatively induced breaks suggested that oxidation of DNA bases was not a dominant mechanism of damage. The present findings highlighted the risk posed to fish populations by the assessed chemicals, jointly or individually, emphasizing the need to define regulatory thresholds for all the formulation components and recommending, in particular, the revision of the hazard classification of POEA.

Species traits as predictors for intrinsic sensitivity of aquatic invertebrates to the insecticide chlorpyrifos

Abstract: Ecological risk assessment (ERA) has followed a taxonomy-based approach, making the assumption that related species will show similar sensitivity to toxicants, and using safety factors or species sensitivity distributions to extrapolate from tested to untested species. In ecology it has become apparent that taxonomic approaches may have limitations for the description and understanding of species assemblages in nature. Therefore it has been proposed that the inclusion of species traits in ERA could provide a useful and alternative description of the systems under investigation. At the same time, there is a growing recognition that the use of mechanistic approaches in ERA, including conceptual and quantitative models, may improve predictive and extrapolative power. Purposefully linking traits with mechanistic effect models could add value to taxonomy-based ERA by improving our understanding of how structural and functional system facets may facilitate inter-species extrapolation. Here, we explore whether and in what ways traits can be linked purposefully to mechanistic effect models to predict intrinsic sensitivity using available data on the acute sensitivity and toxicokinetics of a range of freshwater arthropods exposed to chlorpyrifos. The results of a quantitative linking of seven different endpoints and twelve traits demonstrate that while quantitative links between traits and/or trait combinations and process based (toxicokinetic) model parameters can be established, the use of simple traits to predict classical sensitivity endpoints yields little insight. Remarkably, neither of the standard sensitivity values, i.e. the LC50 or EC50, showed a strong correlation with traits. Future research in this area should include a quantitative linking of toxicodynamic parameter estimations and physiological traits, and requires further consideration of how mechanistic trait-process/parameter links can be used for prediction of intrinsic sensitivity across species for different substances in ERA.

Toxicokinetic-toxicodynamic modelling of survival of Gammarus pulex in multiple pulse exposures to propiconazole: model assumptions, calibration data requirements and predictive power

Abstract: Toxicokinetic-toxicodynamic (TKTD) models quantify the time-course of internal concentration, which is defined by uptake, elimination and biotransformation (TK), and the processes which lead to the toxic effects (TD). TKTD models show potential in predicting pesticide effects in fluctuating concentrations, but the data requirements and validity of underlying model assumptions are not known. We calibrated TKTD models to predict survival of Gammarus pulex in propiconazole exposure and investigated the data requirements. In order to assess the need of TK in survival models, we included or excluded simulated internal concentrations based on pre-calibrated TK. Adding TK did not improve goodness of fits. Moreover, different types of calibration data could be used to model survival, which might affect model parameterization. We used two types of data for calibration: acute toxicity (standard LC50, 4 d) or pulsed toxicity data (total length 10 d). The calibration data set influenced how well the survival in the other exposure scenario was predicted (acute to pulsed scenario or vice versa). We also tested two contrasting assumptions in ecotoxicology: stochastic death and individual tolerance distribution. Neither assumption fitted to data better than the other. We observed in 10-d toxicity experiments that pulsed treatments killed more organisms than treatments with constant concentration. All treatments received the same dose, i.e. the time-weighted average concentration was equal. We studied mode of toxic action of propiconazole and it likely acts as a baseline toxicant in G. pulex during 10-days of exposure for the endpoint survival.

Surface-functionalization effects on uptake of fluorescent polystyrene nanoparticles by model biofilms

Abstract: A study was conducted to investigate the role of nanoparticle (NP) surface functionalization/charge on their uptake by biofilms. Biofilms, bacterial colonies attached to surfaces via extracellular polymers, are effec- tive at removing suspended nanomaterials from the aque- ous phase. However, the mechanisms regulating particle uptake are unknown. Here, it was shown that the mecha- nism was strongly dependent on the nanoparticle surface ionization, and not the core composition of the NP. Uptake experiments were conducted using laboratory-cultured biofilms. The biofilms were incubated in the presence of fluorescent polystyrene NPs with either negatively-charged surfaces (i.e. functionalized with sulfated (SO4 - -NP) or carboxylated (COO - -NP) groups) or positively-charged surfaces (functionalized with primary amines, Amine-P). Particles with negatively-charged sulfated surfaces associ- ated most strongly to biofilms across all experimental conditions. Associations of positively-charged amine particles with biofilms were greatest at high ionic condi- tions resembling those of seawater, but were sensitive to changes in ionic strength. Sorption of COO - -NPs was lowest, relative to other particle types, and was not sensi- tive to ionic strength. The results of this study support an emerging precedent that biofilms may be an effective player in the binding and sequestration of nanoparticles in aqueous systems.

Analyses of gene expression and physiological changes in Microcystis aeruginosa reveal the phytotoxicities of three environmental pollutants

Abstract: When the concentrations of ampicillin (Amp), atrazine (Atr) and cadmium chloride (Cd) reach excessive quantities, they become toxic to aquatic organisms. Due to the acceleration of the industrialization and the intensification of human activities, the incidence and concentrations of these types of pollutants in aquatic systems are increasing. The primary purpose of this study was to evaluate the short-term effects of Amp, Atr and Cd on the physiological indices and gene expression levels in Microcystis aeruginosa. These three pollutants significantly induced antioxidant activity but continuously accelerated the cellular oxidative damage in microalgae, which suggests an imbalance between the oxidant and the antioxidant systems. Amp, Atr and Cd also decreased the transcription of psaB, psbD1 and rbcL; the lowest transcription of these genes was only 38.1, 23.7 and 7% of the control, respectively. These three pollutants affected nitrogen (N) and phosphorous (P) uptake by inhibiting the transcription of N or P absorbing and transporting related genes, and they down regulated the transcription of microcystin-related genes, which caused a decrease of microcystin levels; and the lowest level of microcystin was only 42.4% of the control. Our results suggest that these pollutants may cause pleiotropic effects on algal growth and physiological and biochemical reactions, and they may even affect secondary metabolic processes.

Single and combined effects of selected pharmaceuticals at sublethal concentrations on multiple biomarkers in Carassius auratus

Abstract: In this study, the sublethal effects of caffeine, sulfamethoxazole and their mixture on goldfish (Carassius auratus) were investigated, the biomarkers including acetylcholinesterase (AChE) in brain, 7-ethoxyresorufin O-deethylase (EROD), glutathione S-transferase (GST) and superoxide dismutase (SOD) in liver and vitellogenin (VTG) in serum were determined after 1, 2, 4, and 7 days of exposure. AChE activity was significantly inhibited by caffeine (≥0.4 mg/l), sulfamethoxazole (≥0.4 mg/l) and their mixtures (≥0.048 mg/l) during all exposure periods, and obvious concentration-response and time-response relationships were obtained. EROD, GST and SOD activities were significantly increased by individual compounds and mixtures in most cases. GST induction exhibited bell-shaped concentration-response curves. Serum VTG was significantly induced by 2 mg/l of caffeine, 10 mg/l of sulfamethoxazole and the mixtures at concentrations ≥1.2 mg/l. In general, the two pharmaceuticals induced similar biological responses. The joint effect of caffeine/sulfamethoxazole was additive with regard to AChE and GST activity variation and was antagonistic with regard to EROD and SOD induction. The results indicated that multiple biomarker response method might be a useful tool for describing an integrated toxicological effect of chemicals. VTG induction suggested that caffeine and sulfamethoxazole may cause a slightly feminization effect.

Evaluating the ameliorative effect of natural dissolved organic matter (DOM) quality on copper toxicity to Daphnia magna : improving the BLM

Abstract: Various quality predictors of seven different natural dissolved organic matter (DOM) and humic substances were evaluated for their influence on protection of Daphnia magna neonates against copper (Cu) toxicity. Protection was examined at 3 and 6 mg l−1 of dissolved organic carbon (DOC) of each DOM isolate added to moderately hard, dechlorinated water. Other water chemistry parameters (pH, concentrations of DOC, calcium, magnesium and sodium) were kept relatively constant. Predictors included absorbance ratios Abs254/365 (index of molecular weight) and Abs-octanol254/Abs-water254 (index of lipophilicity), specific absorption coefficient (SAC340; index of aromaticity), and fluorescence index (FI; index of source). In addition, the fluorescent components (humic-like, fulvic-like, tryptophan-like, and tyrosine-like) of the isolates were quantified by parallel factor analysis (PARAFAC). Up to 4-fold source-dependent differences in protection were observed amongst the different DOMs. Significant correlations in toxicity amelioration were found with Abs254/365, Abs-octanol254/Abs-water254, SAC340, and with the humic-like fluorescent component. The relationships with FI were not significant and there were no relationships with the tryptophan-like or tyrosine-like fluorescent components at 3 mg C l−1, whereas a negative correlation was seen with the fulvic-like component. In general, the results indicate that larger, optically dark, more lipophilic, more aromatic DOMs of terrigenous origin, with higher humic-like content, are more protective against Cu toxicity. A method for incorporating SAC340 as a DOM quality indicator into the Biotic Ligand Model is presented; this may increase the accuracy for predicting Cu toxicity in natural waters.

Direct and indirect effects of the glyphosate formulation Glifosato Atanor® on freshwater microbial communities.

Abstract: Glyphosate-based formulations are among the most widely used herbicides in the world. The effect of the formulation Glifosato Atanor® on freshwater microbial communities (phytoplankton, bacterioplankton, periphyton and zooplankton) was assessed through a manipulative experiment using six small outdoor microcosms of small volume. Three of the microcosms were added with 3.5 mg l−1 of glyphosate whereas the other three were left as controls without the herbicide. The treated microcosms showed a significant increase in total phosphorus, not fully explained by the glyphosate present in the Glifosato Atanor®. Therefore, part of the phosphorus should have come from the surfactants of the formulation. The results showed significant direct and indirect effects of Glifosato Atanor® on the microbial communities. A single application of the herbicide caused a fast increase both in the abundance of bacterioplankton and planktonic picocyanobacteria and in chlorophyll a concentration in the water column. Although metabolic alterations related to oxidative stress were induced in the periphyton community, the herbicide favored its development, with a large contribution of filamentous algae typical of nutrient-rich systems, with shallow and calm waters. An indirect effect of the herbicide on the zooplankton was observed due to the increase in the abundance of the rotifer Lecane spp. as a consequence of the improved food availability given by picocyanobacteria and bacteria. The formulation affected directly a fraction of copepods as a target. It was concluded that the Glifosato Atanor® accelerates the deterioration of the water quality, especially when considering small-volume water systems.

Sub-lethal effects induced by a mixture of three non-steroidal anti-inflammatory drugs (NSAIDs) on the freshwater bivalve Dreissena polymorpha

Abstract: Non-steroidal anti-inflammatory drugs (NSAIDs) are the sixth top-selling drugs worldwide and are commonly found in freshwater ecosystems in the high ng/l to low μg/l range. Recent studies have investigated both the acute and the chronic toxicity of single NSAIDs on different biological models, but these studies have completely neglected the fact that, in the environment, non-target organisms are exposed to mixtures of drugs that have unforeseeable toxicological behavior. This work investigated the sub-lethal effects induced by a mixture of three common NSAIDs, namely, diclofenac, ibuprofen and paracetamol, on the freshwater bivalve, the zebra mussel (Dreissena polymorpha). The mussels were exposed to three different environmental concentrations of the mixture (Low, Mid and High). A multi-biomarker approach was used to highlight cyto-genotoxic effects and the imbalance of the oxidative status of the treated specimens. The Neutral Red Retention Assay (NRRA) was used as a biomarker of cytotoxicity, whereas the activities of catalase, superoxide dismutase, glutathione peroxidase and glutathione S-transferase were measured to assess the role played by the oxidative stress enzymes. In addition, the single cell gel electrophoresis assay, the DNA Diffusion assay and the micronucleus test were used to investigate possible genotoxic effects. According to our NRRA results, each treatment was able to induce a significant cellular stress in bivalves, probably due to the raise of oxidative stress, as indicated by the alteration of enzyme activities measured in treated specimens. Moreover, the mixture induced significant enhancements of DNA fragmentation, which preluded fixed genetic damage, as highlighted by the increase of both apoptotic and micronucleated cells.

Differences in susceptibility of five cladoceran species to two systemic insecticides, imidacloprid and fipronil.

Abstract: Differences in susceptibility of five cladocerans to the neonicotinoid imidacloprid and the phenyl-pyrazole fipronil, which have been dominantly used in rice fields of Japan in recent years, were examined based on short-term (48-h), semi-static acute immobilization exposure tests. Additionally, we compared the species sensitivity distribution (SSD) patterns of both insecticides between two sets of species: the five tested cladocerans and all other aquatic organisms tested so far, using data from the ECOTOX database of U.S. Environmental Protection Agency (USEPA). The sensitivity of the test species to either imidacloprid or fipronil was consistent, spanning similar orders of magnitude (100 times). At the genus level, sensitivities to both insecticides were in the following descending order: Ceriodaphnia > Moina > Daphnia. A positive relationship was found between body lengths of each species and the acute toxicity (EC50) of the insecticides, in particular fipronil. Differences in SSD patterns of imidacloprid were found between the species groups compared, indicating that test cladocerans are much less susceptible than other aquatic species including amphibians, crustaceans, fish, insects, mollusks and worms. However, the SSD patterns for fipronil indicate no difference in sensitivity between cladocerans tested and other aquatic organisms despite the greater exposure, which overestimates the results, of our semi-static tests. From these results, Ceriodaphnia sp. should be considered as more sensitive bioindicators (instead of the standard Daphnia magna) for ecotoxicological assessments of aquatic ecosystems. In addition, we propose that ecotoxicity data associated with differences in susceptibility among species should be investigated whenever pesticides have different physicochemical properties and mode of action.

Analyses of n-alkanes degrading community dynamics of a high-temperature methanogenic consortium enriched from production water of a petroleum reservoir by a combination of molecular techniques

Abstract: Despite the knowledge on anaerobic degradation of hydrocarbons and signature metabolites in the oil reservoirs, little is known about the functioning microbes and the related biochemical pathways involved, especially about the methanogenic communities. In the present study, a methanogenic consortium enriched from high-temperature oil reservoir production water and incubated at 55 °C with a mixture of long chain n-alkanes (C15–C20) as the sole carbon and energy sources was characterized. Biodegradation of n-alkanes was observed as methane production in the alkanes-amended methanogenic enrichment reached 141.47 μmol above the controls after 749 days of incubation, corresponding to 17 % of the theoretical total. GC–MS analysis confirmed the presence of putative downstream metabolites probably from the anaerobic biodegradation of n-alkanes and indicating an incomplete conversion of the n-alkanes to methane. Enrichment cultures taken at different incubation times were subjected to microbial community analysis. Both 16S rRNA gene clone libraries and DGGE profiles showed that alkanes-degrading community was dynamic during incubation. The dominant bacterial species in the enrichment cultures were affiliated with Firmicutes members clustering with thermophilic syntrophic bacteria of the genera Moorella sp. and Gelria sp. Other represented within the bacterial community were members of the Leptospiraceae, Thermodesulfobiaceae, Thermotogaceae, Chloroflexi, Bacteroidetes and Candidate Division OP1. The archaeal community was predominantly represented by members of the phyla Crenarchaeota and Euryarchaeota. Corresponding sequences within the Euryarchaeota were associated with methanogens clustering with orders Methanomicrobiales, Methanosarcinales and Methanobacteriales. On the other hand, PCR amplification for detection of functional genes encoding the alkylsuccinate synthase α-subunit (assA) was positive in the enrichment cultures. Moreover, the appearance of a new assA gene sequence identified in day 749 supported the establishment of a functioning microbial species in the enrichment. Our results indicate that n-alkanes are converted to methane slowly by a microbial community enriched from oilfield production water and fumarate addition is most likely the initial activation step of n-alkanes degradation under thermophilic methanogenic conditions.

Differences in ecological impacts of systemic insecticides with different physicochemical properties on biocenosis of experimental paddy fields

Abstract: The environmental risks of pesticides are typically determined by laboratory single-species tests based on OECD test guidelines, even if biodiversity should also be taken into consideration. To evaluate how realistic these assessments are, ecological changes caused by the systemic insecticides imidacloprid and fipronil, which have different physicochemical properties, when applied at recommended commercial rates on rice fields were monitored using experimental paddy mesocosms. A total of 178 species were observed. There were no significant differences in abundance of crop arthropods among the experimental paddies. However, zooplankton, benthic and neuston communities in imidacloprid-treated field had significantly less abundance of species than control and fipronil fields. Significant differences in abundance of nekton community were also found between both insecticide-treated paddies and control. Influences on the growth of medaka fish were also found in both adults and their fries. Both Principal Response Curve analysis (PRC) and Detrended Correspondence Analysis (DCA) showed the time series variations in community structure among treatments, in particular for imidacloprid during the middle stage of the experimental period. These results show the ecological effect-concentrations (LOEC ~ 1 μg/l) of these insecticides in mesocosms, especially imidacloprid, are clearly different from their laboratory tests. We suggest that differences in the duration of the recovery process among groups of species are due to different physicochemical properties of the insecticides. Therefore, realistic prediction and assessment of pesticide effects at the community level should consider not only the sensitivity traits and interaction among species but also the differences in physicochemical characteristics of each pesticide.

Antimicrobial effects of commercial silver nanoparticles are attenuated in natural streamwater and sediment

Abstract: Given the demonstrated antimicrobial properties of silver nanoparticles (AgNPs), and the key role that microorganisms play in performing critical ecosystem functions such as decomposition and nutrient cycling, there is growing concern that AgNP pollution may negatively impact ecosystems. We examined the response of streamwater and sediment microorganisms to commercially available 21 ± 17 nm AgNPs, and compared AgNP impacts to those of dissolved-Ag added as AgNO3. We show that in streamwater, AgNPs and AgNO3 decreased respiration in proportion to dissolved-Ag concentrations at the end of the incubation (r2 = 0.78), while in sediment the only measurable effect of AgNPs was a 14 % decrease in sulfate concentration. This contrasts with the stronger effects of dissolved-Ag additions in both streamwater and sediment. In streamwater, addition of dissolved-Ag at a level equivalent to the lowest AgNP dose led to respiration below detection, a 55 % drop in phosphatase enzyme activity, and a 10-fold increase in phosphate concentration. In sediment, AgNO3 addition at a level equivalent to the highest AgNP addition led to a 34 % decrease in respiration, a 55 % increase in microbial biomass, and a shift in bacterial community composition. The results of this study suggest that, in similar freshwater environments, the short-term biological impacts of AgNPs on microbes are attenuated by the physical and chemical properties of streamwater and sediment.

Acute toxicity and responses of antioxidant systems to 1-methyl-3-octylimidazolium bromide at different developmental stages of goldfish

Abstract: Acute toxicity of 1-methyl-3-octylimidazolium bromide ([C8mim]Br) to goldfish at different developmental stages and responses of the antioxidant system in adult goldfish were evaluated in the present study. The results indicate that post-embryonic developmental toxicity of [C8mim]Br on goldfish is developmental-stage dependent. The juvenile and larva goldfish are more sensitive to [C8mim]Br-toxicity than the adult fish. Histological observations in adult goldfish reveal that acute [C8mim]Br exposure damages the hepatopancreas, intestines, and kidneys, indicating that these are possible target organs of [C8mim]Br toxicity in goldfish. Subsequent biochemical assays in adult goldfish show that [C8mim]Br also induces changes in the activities of the superoxide dismutase, catalase, glutathione peroxidase, and glutathione content of fish hepatopancreas. These results suggest that [C8mim]Br exposure may induce oxidant stress and lipid peroxidation in hepatopancreas of adult goldfish. In addition, we also find that [C8mim]Br causes a remarkable increase in malondialdehyde (MDA) levels in the hepatopancreas of adult goldfish, and thus we think that the MDA level change can be a biomarker of [C8mim]Br toxicity in goldfish. The present study indicates that ionic liquids can be a threat to the survival, growth, and development of the fish population once they are accidentally leaked into aquatic ecosystems.

Toxicity and genotoxicity of organic and inorganic nanoparticles to the bacteria Vibrio fischeri and Salmonella typhimurium.

Abstract: The present work aimed at evaluating the toxicity and genotoxicity of two organic (vesicles composed of sodium dodecyl sulphate/didodecyl dimethylammonium bromide—SDS/DDAB and of monoolein and sodium oelate—Mo/NaO) and four inorganic (titanium oxide—TiO2, silicon titanium—TiSiO4, Lumidot™-CdSe/ZnS, and gold nanorods) nanoparticles (NP), suspended in two aqueous media (Milli Q® water and American Society for Testing and Materials (ASTM) hardwater), to the bacteria Vibrio fischeri (Microtox® test) and Salmonella typhimurium-his − (Ames® test with strains TA98 and TA100). Aiming a better understanding of these biological responses physical and chemical characterization of the studied NP suspensions was carried out. Results denoted a high aggregation state of the NP in the aqueous suspensions, with the exception of SDS/DDAB and Mo/NaO vesicles, and of nanogold suspended in Milli Q water. This higher aggregation was consistent with the low values of zeta potential, revealing the instability of the suspensions. Regarding toxicity data, except for nano TiO2, the tested NP significantly inhibited bioluminescence of V. fischeri. Genotoxic effects were only induced by SDS/DDAB and TiO2 for the strain TA98. A wide range of toxicity responses was observed for the six tested NP, differing by more than 5 orders of magnitude, and suggesting different modes of action of the tested NP.

Effects of lead on the growth, lead accumulation and physiological responses of Pluchea sagittalis

Abstract: This work aimed to study the process of stress adaptation in root and leaves of different developmental stages (apex, middle and basal regions) of Pluchea sagittalis (Lam.) Cabrera plants grown under exposure to five Pb levels (0, 200, 400, 600 and 1000 μM) for 30 days. Pb concentration and content in roots, stems, and leaves of different developmental stages increased with external Pb level. Consumption of nutrient solution, transpiration ratio, leaf fresh weight, leaf area, and shoot length decreased upon addition of Pb treatments. However, dry weight of shoot parts and roots did not decrease upon addition of Pb treatments. Based on index of tolerance, the roots were much more tolerant to Pb than shoots. δ-aminolevulinic acid dehydratase activity was decreased by Pb treatments, whereas carotenoid and chlorophyll concentrations were not affected. Lipid peroxidation and hydrogen peroxide concentration both in roots and leaves increased with increasing Pb levels. Pb treatments increased ascorbate peroxidase activity in all plant parts, while superoxide dismutase activity increased in leaves and did not change in roots. Catalase activity in leaves from the apex shoot was not affected by Pb, but in other plant parts it was increased. Pb toxicity caused increase in non-protein thiol groups concentration in shoot parts, whereas no significant difference was observed in roots. Both root and shoot ascorbic acid concentration increased with increasing Pb level. Therefore, it seems that Pb stress triggered an efficient defense mechanism against oxidative stress in P. sagittalis but its magnitude was depending on the plant organ and of their physiological status. In addition, these results suggest that P. sagittalis is Pb-tolerant. In conclusion, P. sagittalis is able to accumulate on average 6730 and 550 μg Pb g−1 dry weight, respectively, in the roots and shoot, a physiological trait which may be exploited for the phytoremediation of contaminated soils and waters.

Interactions of silver nanoparticles with the marine macroalga, Ulva lactuca.

Abstract: The marine macroalga, Ulva lactuca, has been exposed for 48 h to different concentrations of Ag added as either silver nanoparticles (AgNP) or aqueous metal (AgNO3) and the resulting toxicity, estimated from reductions in quenching of chlorophyll-a fluorescence, and accumulation of Ag measured. Aqueous Ag was toxic at available concentrations as low as about 2.5 μg l−1 and exhibited considerable accumulation that could be defined by the Langmuir equation. AgNP were not phytotoxic to the macroalga at available Ag concentrations up to at least 15 μg l−1 and metal measured in U. lactuca was attributed to a physical association of nanoparticles at the algal surface. At higher AgNP concentrations, a dose–response relationship was observed that was similar to that for aqueous Ag recorded at much lower concentrations. These findings suggest that AgNP are only indirectly toxic to marine algae through the dissolution of Ag+ ions into bulk sea water, albeit at concentrations orders of magnitude greater than those predicted in the environment.

Biomarker assessment of toxicity with miniaturised bioassays: diclofenac as a case study

Abstract: The development of suitable biomarker-based microbioassays with model species with ecological relevance would help increase the cost-efficiency of routine environmental monitoring and chemical toxicity testing. The anti-inflammatory drug diclofenac has been widely reported in the environment but ecotoxicological data are scarce. The aim of this work is to assess the acute and chronic sublethal toxicity of diclofenac in relevant taxa of aquatic and riparian ecosystems (the fish Danio rerio and the fern Polystichum setiferum). Reliable biomarkers of cell viability (mitochondrial activity), plant physiology (chlorophyll), growth (DNA content) or oxidative damage (lipid peroxidation) were assessed as sensitive endpoints of toxicity. DNA quantification shows that diclofenac induces acute lethal phytotoxicity at 24 and 48 h (LOECs 30 and 0.3 μg l−1, respectively). Hormetic effects in mitochondrial activity in spores of Polystichum setiferum mask lethality, and adverse effects are only observed at 48 h (LOEC 0.3 μg l−1). In chronic exposure (1 week) LOEC for DNA is 0.03 μg l−1. Mitochondrial activity shows a strong hormetic stimulation of the surviving spore population (LOEC 0.3 μg l−1). Little changes are observed in chlorophyll autofluorescence (LOEC 0.3 μg l−1). A very short exposure (90 min) of zebrafish embryos induces a reduction of lipid peroxidation at 0.03 μg l−1. Environmental concentrations of diclofenac can be deleterious for the development of significant populations of sensitive individuals in aquatic and riparian ecosystems.

Pesticide application to agricultural fields: effects on the reproduction and avoidance behaviour of Folsomia candida and Eisenia andrei

Abstract: The objective of this work was to assess the impact of pesticide application to non-target soil organisms simulating what happens following pesticide application in agricultural fields and thus obtaining higher realism on results obtained. For that purpose, three commercial formulations containing the insecticides chlorpyrifos and endosulfan and the herbicide glyphosate were applied to a Mediterranean agricultural field. The soil was collected after spraying and dilution series were prepared with untreated soil to determine the impact of the pesticides on the avoidance behaviour and reproduction of the earthworm Eisenia andrei and the collembolan Folsomia candida. A significant avoidance was observed at the recommended field dose in case of endosulfan by earthworms (60 %) and in case of chlorpyrifos by collembolans (64 %). In addition, both insecticides affected the number of juveniles produced by the earthworms (EC50 were below the recommended field dose). Glyphosate did not seem to affect either earthworms or collembolans in the recommended field dose. Folsomia candida was more sensitive to pesticide application than Eisenia andrei, what was corroborated by the EC50 and LC50 values. In conclusion, insecticides may affect the structure of the soil community by reducing the survival of collembolans and the reproductive capacity of collembolans and earthworms.