Direct and indirect effects of pesticides on a benthic grazer during its life cycle

Background

Macroinvertebrates in aquatic ecosystems are repeatedly exposed to pesticides during their life cycle. Effects of consecutive exposure during different life stages and possible synergistic effects are not addressed in the standardized hazard assessment. The present study investigated two environmentally relevant exposure scenarios in batch (microcosm) and artificial indoor stream (mesocosm) experiments using the larvae of the mayfly Rhithrogena semicolorata (grazer) and natural aufwuchs. Grazers were analysed regarding growth, physiological condition, and drift behaviour, while the aufwuchs was analysed in terms of biomass using the particulate organic carbon as well as the chlorophyll a content. The aim was to reveal direct and indirect effects of an herbicide exposure during autumn on juvenile grazers and an insecticide exposure during spring on semi-juvenile grazers.

Results

Direct and indirect effects were found in both exposure scenarios at environmentally relevant concentrations. In the herbicide exposure scenario with terbutryn, clear direct effects on the aufwuchs community with a LOEC of 0.38 µg L^?1 were found. Effect levels of grazers due to indirect effects were equal, with the overnight drift being the most sensitive grazer endpoint. In the insecticide exposure scenario, clear lethal and sub lethal effects of lambda-cyhalothrin were evident. Derived LC₅₀ values for the artificial indoor stream and batch experiment were 2.42 µg g^?1 OC (69 days) and 1.2 µg g^?1 OC (28 days), respectively. Sub lethal effects in terms of increased drift as well-reduced growth and triglyceride levels were found at concentrations of 1.4 and 0.09 µg g^?1 OC (LOECs). These results were confirmed by the batch experiment, which revealed effect values in the similar range. Finally, a clear indirect effect of the insecticide on the aufwuchs was evident in the batch experiment with an LOEC at 0.9 µg g^?1 OC.

Conclusion

Toxicity Exposure Ratios calculated with the derived effect values indicate a risk for the investigated grazer by both pesticides. Moreover, observed indirect effects during the herbicide exposure seem to be able to affect the grazers during a second exposure with an insecticide, due to reduced physiological conditions. We suggest further research with time-shifted exposure scenarios to gain a better understanding of the complex interactions of pesticides with the life cycle and the food webs of macroinvertebrates.

     

10种常用农药对赤子爱胜蚓(Eisenia foetida)的急性毒性效应

为了评价农药对土壤无脊椎动物种群的生态风险,采用OECD标准滤纸法和人工土壤法测定了10种常用农药对蚯蚓(Eisenia foetida)的急性毒性效应。采用滤纸法,48h测定结果表明,4种氨基甲酸酯类(异丙威、甲萘威、速灭威和丁硫克百威)和2种有机磷类(毒死蜱和哒嗪硫磷)农药对蚯蚓的急性毒性(其LC50值为3.50(2.77～4.44)～72.42(59.58～88.05)μg·cm-2)明显高于3种昆虫生长调节剂(噻嗪酮、虫酰肼和呋喃虫酰肼)和吡蚜酮对蚯蚓的急性毒性(其LC50值>629.1μg·cm-2)。采用人工土壤法,14d测定结果表明,上述4种氨基甲酸酯类农药对蚯蚓的急性毒性(其LC50值为59.61(55.13～64.44)～134.1(127.0～141.5)mg·kg-1)明显高于上述2种有机磷类农药和其他农药对蚯蚓的急性毒性(其LC50值为193.0(180.1～206.8)～386.4(359.7～414.2)mg·kg-1)。上述结果表明,不同类型的农药对蚯蚓的毒性存在较大差异,且同一类型的不同农药品种对蚯蚓的毒性也存在较大差异。总体来看,氨基甲酸酯类农药比其他类型的农药对蚯蚓具有更高的毒性。根据《化学农药环境安全评价试验准则》,本研究采用人工土壤法测定的所有农药对蚯蚓均为低毒级。     

New effects of Roundup on amphibians: predators reduce herbicide mortality; herbicides induce antipredator morphology

The use of pesticides is important for growing crops and protecting human health by reducing the prevalence of targeted pest species. However, less attention is given to the potential unintended effects on nontarget species, including taxonomic groups that are of current conservation concern. One issue raised in recent years is the potential for pesticides to become more lethal in the presence of predatory cues, a phenomenon observed thus far only in the laboratory. A second issue is whether pesticides can induce unintended trait changes in nontarget species, particularly trait changes that might mimic adaptive responses to natural environmental stressors. Using outdoor mesocosms, I created simple wetland communities containing leaf litter, algae, zooplankton, and three species of tadpoles (wood frogs [Rana sylvatica or Lithobates sylvaticus], leopard frogs [R. pipiens or L. pipiens], and American toads [Bufo americanus or Anaxyrus americanus]). I exposed the communities to a factorial combination of environmentally relevant herbicide concentrations (0, 1, 2, or 3 mg acid equivalents [a.e.]/L of Roundup Original MAX) crossed with three predator-cue treatments (no predators, adult newts [Notophthalmus viridescens], or larval dragonflies [Anax junius]). Without predator cues, mortality rates from Roundup were consistent with past studies. Combined with cues from the most risky predator (i.e., dragonflies), Roundup became less lethal (in direct contrast to past laboratory studies). This reduction in mortality was likely caused by the herbicide stratifying in the water column and predator cues scaring the tadpoles down to the benthos where herbicide concentrations were lower. Even more striking was the discovery that Roundup induced morphological changes in the tadpoles. In wood frog and leopard frog tadpoles, Roundup induced relatively deeper tails in the same direction and of the same magnitude as the adaptive changes induced by dragonfly cues. To my knowledge, this is the first study to show that a pesticide can induce morphological changes in a vertebrate. Moreover, the data suggest that the herbicide might be activating the tadpoles' developmental pathways used for antipredator responses. Collectively, these discoveries suggest that the world's most widely applied herbicide may have much further-reaching effects on nontarget species than previous considered.     

新烟碱类农药的污染现状及转化行为研究进展

新烟碱类(neonicotinoids)农药自在20世纪80年代中期引入以来,便发展成为世界上使用最广泛的一类杀虫剂。近年来,该类化合物在环境中被不断检出,并通过饮用水和食物等方式进入食物链,给人类健康带来极大的安全隐患。文章综述了新烟碱类杀虫剂在空气、土壤、水相、生物体和人体中的暴露浓度,以及这类农药在环境中的转化行为。目前针对新烟碱类杀虫剂的环境暴露研究仍十分有限,需要更全面调查其在多环境介质中的暴露状况以综合评价其生态风险。关于新烟碱类化合物的毒理学研究主要集中在急性毒性试验,而对于其他形式的毒性效应有待进一步研究。最后对新烟碱农药未来的研究重点和方向提出了展望。     

基于RICEWQ—EXAMS模型的东苕溪流域稻田用药的水生生态及健康风险评价

构建了东苕溪流域水稻地表水暴露场景,对国外已广泛应用的稻田-地表水暴露评价模型（RICEWQ—EX—AMS）进行二次开发,并应用构建的场景和开发的模型对东苕溪流域稻田常用农药品种进行水生生态风险评价和健康风险评价。结果表明,在所评价的10种东苕溪流域常用农药品种中,除草剂氟乐灵对藻具有急性高风险,杀虫剂硫丹对鱼既具有急性高风险,又具有慢性风险,阿维菌素对泾具有急性高风险,氟铃脲对涵既具有急性高风险,又具有慢性风险,其余品种对鱼、溢和藻的急慢性风险均较低。运用传统评价法和风险评价模型计算法得出了相一致的结论,即所评价的10个农药品种对人体健康均无风险,该评价结果与所评价农药的实际风险表现较吻合。认为所构建的场景、模型能较好地用于东苕溪流域稻田农药品种的风险评价。     

Toxicity potential of Grassate® a non-selective herbicide on snails (Achachatina marginata) and earthworms (Aporrectodea longa)

The application of herbicides in agricultural practices may pose certain toxic effects on non-target species including molluscs (snails), earthworms and other soil-dwelling organisms, which would in turn put the lives of humans at great risk. This study considered the lethal and sublethal effects of Grassate®, a non-selective glyphosate-based herbicide on snails (Archachatina marginata) and earthworms (Aporrectodea longa) with regards to ecotoxicological risk assessment. The lethal concentration LC₅₀ for the test chemical averaged at 1.731?±?0.05 and 3.045?±?0.08?mg?kg^?1 for snails and earthworms, respectively. There was decrease in growth and biomass with increased concentration for the species. Growth inhibition of 11–38% in size and 7–43% in length was obtained for earthworms while 26–65% reduction in weight was reported for snails. Using the ecotoxicological risk assessment matrix, the herbicide was classified as D 4 (A; P; E) or 16 (A; P; E), which could be considered as high risk to animals, plant and the environment. Thus, if these herbicides are used uncontrollably and unregulated, further consequence could lead to harmful effects on humans who feed on snails, a rich source of protein and depend on earthworms for nutrient.     

Cytochrome P450 monooxygenase specific activity reduction in wheat <Emphasis Type="Italic">Triticum aestivum</Emphasis> induced by soil roxithromycin stress

Roxithromycin, as widely used medicine and livestock growth promoter, arouses concern because its occurrence and persistence in soil environments. However, effects of roxithromycin in higher plants are still vague. Accordingly, we hypothesized that roxithromycin-contaminated soil may exhibits ecotoxicological effects in wheat (Triticum aestivum). In this study, effects induced by a gradient concentration of roxithromycin stress (0.01, 0.1, 1, 10, and 100 mg·kg^–1) was investigated in a 7-d soil test in T. aestivum. Results indicated that the specific activity of cytochrome P450 (CYP450) monooxygenase was decreased dramatically with the concentration of roxithromycin in soil. The IC₅₀ value was 8.78 mg·kg^–1 of roxithromycin. On the contrary, the growth related endpoints (i.e., the germination percentage, the biomass and the height), the content related endpoints (i.e., soluble protein content and CYP450 content), and the superoxide dismutase (SOD) activity failed to reveal the roxithromycin-induced effects. Further analysis revealed that the CYP450 monooxygenase specific activity reduction was enzymatic mechanism mediated, other than oxidative stress induced. We conclude that the soil roxithromycin declined the CYP450 monooxygenase activity in T. aestivum by the inhibition of the enzymatic mechanism. Further efforts can include, but are not limited to, investigation of joint effects induced by combined exposure of roxithromycin and the pesticides and evaluation of the similar effects in other higher plants.     

Wirkung von Isoproturon auf Laich und Larven zweier Unkenarten (Bombina spec.)

In intensively used arable areas, a contamination of the reproduction ponds with pesticides probably impairs the development of spawn and tadpoles of amphibians, based on the coincidental space and time. Therefore, the effects of the herbicide isoproturon (IPU) on the early life stages of the firebellied toad (Bombina bombina) and the closely related yellowbellied toad (Bombina variegata) were investigated. The results of the exposure with¹⁴C-labelled IPU (1 μg/L) indicated an uptake into the spawn and tadpoles of bothBombina species. The jelly capsules could not protect the embryo from effects of the herbicide. Tadpoles with complete opercula and without external gills were most sensitive to contamination by IPU. Physical and behavioral abnormalities of the tadpoles developed at concentrations, of 0.1 μg/L after 24 h exposure. At increasing IPU-concentrations the number of impaired and dead tadpoles increased significantly compared to the control. The enzymatic system of the Glutathion S-Transferase (GST) of theBombina tadpoles were influenced significantly by the duration and concentration of IPU exposure. Compared to the pure active ingredient IPU, the commercial herbicide TOLKAN FLO^® provoked a stronger enzymatic response in the tadpoles. This could be caused by the presence of an emulsifier used in the TOLKAN FLO^® formulation which enhanced the availability of IPU and/or the interaction between IPU and the emulsifier.     

Mitigation of agricultural nonpoint-source pesticide pollution in artificial wetland ecosystems

Contamination caused by pesticides in agriculture is a source of environmental poor water quality in some of the European Union countries. Without treatment or targeted mitigation, this pollution is diffused in the environment. Pesticides and some metabolites are of increasing concern because of their potential impacts on the environment, wildlife and human health. Within the context of the European Union (EU) water framework directive context to promote low pesticide-input farming and best management practices, the EU LIFE project ArtWET assessed the efficiency of ecological bioengineering methods using different artificial wetland (AW) prototypes throughout Europe. We optimized physical and biological processes to mitigate agricultural nonpoint-source pesticide pollution in artificial wetland ecosystems. Mitigation solutions were implemented at full-scale demonstration and experimental sites. We tested various bioremediation methods at seven experimental sites. These sites involved (1) experimental prototypes, such as vegetated ditches, a forest microcosm and 12 wetland mesocosms, and (2) demonstration prototypes: vegetated ditches, three detention ponds enhanced with technology of constructed wetlands, an outdoor bioreactor and a biomassbed. This set up provides a variety of hydrologic conditions, with some systems permanently flooded and others temporarily flooded. It also allowed to study the processes both in field and controlled conditions. In order to compare the efficiency of the wetlands, mass balances at the inlet and outlet of the artificial wetland will be used, taking into account the partition of the studied compound in water, sediments, plants, and suspended solids. The literature background necessary to harmonize the interdisciplinary work is reviewed here and the theoretical framework regarding pesticide removal mechanisms in artificial wetland is discussed. The development and the implementation of innovative approaches concerning various water quality sampling strategies for pesticide load estimates during flood, specific biological endpoints, innovative bioprocess applied to herbicide and copper mitigation to enhance the pesticide retention time within the artificial wetland, fate and transport using a 2D mixed hybrid finite element model are introduced. These future results will be useful to optimize hydraulic functioning, e.g., pesticide resident time, and biogeochemical conditions, e.g., dissipation, inside the artificial wetlands. Hydraulic retention times are generally too low to allow an optimized adsorption on sediment and organic materials accumulated in artificial wetlands. Absorption by plants is not either effective. The control of the hydraulic design and the use of adsorbing materials can be useful to increase the pesticides residence time and the contact between pesticides and biocatalyzers. Pesticide fluxes can be reduced by 50–80% when hydraulic pathways in artificial wetlands are optimized by increasing ten times the retention time, by recirculation of water, and by deceleration of the flow. Thus, using a bioremediation method should lead to an almost complete disappearance of pesticides pollution. To retain and treat the agricultural nonpoint-source po a major stake for a sustainable development.     

Effects of chytrid fungus and a glyphosate-based herbicide on survival and growth of wood frogs (Lithobates sylvaticus)

Anthropogenic-derived stressors in the environment, such as contaminants, are increasingly considered important cofactors that may decrease the immune response of amphibians to pathogens. Few studies, however, have integrated amphibian disease and contaminants to test this multiple-stressor hypothesis for amphibian declines. We examined whether exposure to sublethal concentrations of a glyphosate-based herbicide and two strains of the pathogenic chytrid fungus, Batrachochrytrium dendrobatidis (Bd) could: (1) sublethally affect wood frogs (Lithobates sylvaticus) by altering the time to and size at metamorphosis, and (2) directly affect survivability of wood frogs after metamorphosis. Neither Bd strain nor herbicide exposure alone significantly altered growth or time to metamorphosis. The two Bd strains did not differ in their pathogenicity, and both caused mortality in post-metamorphic wood frogs. There was no evidence of an interaction between treatments, indicating a lack of herbicide-induced susceptibility to Bd. However, the trends in our data suggest that exposure of wood frogs to a high concentration of glyphosate-based herbicide may reduce Bd-caused mortality compared to animals exposed to Bd alone. These results exemplify the complexities inherent when populations are coping with multiple stressors. In this case, the perceived stressor, glyphosate-based herbicide, appeared to affect the pathogen more than the host's immune system, relieving the host from disease-caused effects. This suggests caution when invoking multiple stressors as a cause for increased disease susceptibility and indicates that the effects of multiple stressors on disease outcome depend on the interrelationships of stressors to both the pathogen and the host.     

草甘膦对非靶标生物的毒性研究进展

随着现代农业的飞速发展,农药的使用越来越广泛。草甘膦作为全球销量最大的农药除草剂,在自然环境中广泛存在,其对生态环境和人类健康的影响越来越受到人们的关注。文中从水生/两栖动物、土壤生物、哺乳动物以及人群等4个方面对除草剂草甘膦对非靶标生物的毒性研究概况进行综述,并对该领域未来发展趋势做了简要分析与展望,以期为草甘膦的生态安全和环境风险评价提供数据资料,为其合理使用提供理论指导。     

Evidence for DNA-diquat interaction and cytotoxicity in in vitro rat cells

Over 98% of sprayed insecticides and 95% of herbicides reach non-target species in air, soil, and water. Numerous studies have reported that pesticide residues can cause acute and chronic toxicity. Pesticide residues can be carcinogenic, mutagenic, and immunotoxic. There are actually too few studies that bridge the disciplines of chemobioanalysis and environmental toxicology. Here, we assessed the cytotoxicity of a bipyridilium herbicide diquat in rat adrenal pheochromocytoma cells (PC12). Our results show that diquat caused the decrease in cell viability with a lethal concentration 50 (LC₅₀) of 1.4 × 10⁻⁵ mol/L. This cytotoxicity may result from diquat-induced apoptosis, characterized by nuclear fragmentation and chromatin condensation by Hoechst 33324 staining. To explore the possible mechanisms, the interaction between herbicide diquat and calf thymus DNA (ctDNA) was further investigated using fluorescence quenching. The detection of static quenching showed that diquat was linked with ctDNA by electrostatic interaction with a binding constant of 9.288 × 10⁴ L/mol. This is the first study on the interaction of DNA with herbicide diquat by fluorometric method as well as on the evaluation of cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Hoechst staining. Given the widespread use of synthetic pesticides, the data would be valuable for the risk assessment of pesticide residues.     

Entwicklung der Ökotoxikologie von Pflanzenschutzmitteln in den letzten 25 Jahren: Von ersten Diskussionen über Prüforganismen zur probabilistischen Risikoanalyse

In Europe as well as in Germany a remarkable development within the last 25 years took place in the assessment of ecotoxicological effects of Plant Protection Products (PPPs). The starting point was a comparative, retrospective toxicity testing of some selected indicator species. Then, first attempts of a quantitative, prospective hazard and risk assessment with relevant exposure evaluations were made. Currently a probabilistic risk analysis on the landscape level is discussed.Based on personal experiences within the last decades this contribution will highlight the different developments of the ecotoxicological evaluation of PPPs. At first, the selection of suitable test species and the development of suitable standardized test methods was the main task. Thereafter, the discussions focused more on long-term, chronic effects instead of solely acute ones, the development of focused higher tier studies adjusted to better address the specific properties of a PPP and/or the specific concerns resulting from its application, as well as multi-species and ecosystem approaches. With the availability of suitable computer based models a more precise exposure calculation was possible, which allows a more realistic exposure assessment instead of simple worstcase assumptions. Current discussions focus on probabilistic exposure assessments at a local scale.All these developments took place regardless of the insufficient (political) definition of the protection goal and — as a consequence — the need and extent of appropriate safety factors. The development of ecotoxicology was mostly driven by political trends in Europe, mainly by the harmonized registration of PPPs under the still valid Annexes II and III (1996) of the EU Directive 91/414 and the thereby initiated further progression. In the context of this development more and more scientists were offered interesting job opportunities.     

南渡江农业土壤中农药分布特征与生态风险评估

为摸清稻菜轮作土壤中农药(CUPs)残留特征与生态风险,采集海南岛南渡江流域稻菜轮作系统168个土壤样品,利用QuEChERS-超高效液相色谱-串联质谱(UPLC-MS/MS)检测土壤中25种CUPs残留水平,结合土壤因子与作物类型分析CUPs残留关键影响因素,以风险熵法评估其生态风险.结果表明,∑25 CUPs处于0...     

Entwicklung von ökotoxikologischen Instrumenten und ihre rechtliche Implementierung zur marinen ökologischen Risikobewertung von Chemikalien, Pestiziden und Baggergut

Background, aim, and scope The marine environment is often the final sink for pollutants, especially for persistent organic pollutants (POPs) and for persistent, bioaccumulative and toxic (PBT) substances. The body burden of marine top predators in the Baltic Sea, in the North Sea and also in the Arctic seas is alarming. Thus, the question was investigated if the European environmental law considers a prospective marine ecological risk assessment of chemicals, pesticides and dredged material before they are launched on the market or disposed to the sea. Results The analysis of European environmental law showed that the European environmental protection goals demand a good quality status of coastal waters until 2015 (Waterframework Directive) and a good quality status of European seas until 2020 (Marine Strategy Directive), but a specific marine ecological risk assessment of chemicals and pesticides is not sufficiently required in current European legislation. Discussion It was shown that the ecological risk assessment for freshwater ecosystems is, due to the pecularities of pollutant impacts in the marine environment, not adequate to predict marine effects and to protect the marine environment sufficiently. A statistical analysis of international databases on the relative toxicity of narcotics revealed that marine organisms can be significantly more sensitive than freshwater organisms towards substances with an unspecific mode of action (narcotics). Approximately 60?% of the industrial chemicals are classified as narcotics by their mode of action. Thus, this substance class is of environmental importance. Due to the hydrophobic properties and the low solubility of narcotics in seawater, the ecotoxicological assessment of marine sediments was of interest. An estuarine and marine bioassay test set was established and further developed to assess the ecotoxicological potential of brackish and seawater sediments. It was important that the test procedures were adapted to brackish and marine conditions and were harmonised between each other as well as on the international level. Conclusions Beside two bioassays for the ecotoxicological assessment of elutriates of marine and brackish sediments (bacteria bioluminescence test and marine algae test), the implementation and further development of the whole sediment bioassay with the marine amphipod Corophium volutator was important for enhancing the risk assessment. In order to gain a more standardised, all-season available test organism, the marine amphipod was for the first time reproduced under laboratory conditions the whole year round (also in winter), which is the essential basis for the urgently needed chronic whole sediment bioassay. The results of this investigation were implemented in the international (ISO), European (EN) and national (DIN) standardisations. Therefore, a standardised test set is ready for the implementation in the marine ecological risk assessment of chemicals, pesticides and dredged material in international, European and national legislation. Recommendations and perspectives Recommendations to improve the implementation of a marine risk assessment in European regulations are given with the goal to reach the internationally required objective of a sustainable development of the seas.     

林丹和呋喃丹对赤子爱胜蚓存活、生长和繁殖能力的影响

采用人工土培养法,通过急性和亚急性暴露实验,研究了林丹和呋喃丹对赤子爱胜蚓(Eisenia foetida)存活、生长和繁殖能力的影响.结果表明,1)林丹对蚯蚓14d LC50为162.09mg·kg-1,属低等毒性;呋喃丹对蚯蚓14d LC50为3.11mg·kg-1,属中等毒性.2)林丹在急性暴露期显著抑制蚯蚓的生长,在亚急性暴露期低浓度林丹对蚯蚓生长影响不显著,高浓度林丹则显著抑制蚯蚓的生长;呋喃丹在急性和亚急性暴露期对蚯蚓的生长均具有极显著的抑制作用.3)林丹和呋喃丹均可显著抑制蚯蚓的繁殖能力.4)林丹和呋喃丹均可对蚯蚓皮肤结构造成损伤,林丹的损伤程度较严重,呋喃丹相对较弱.5)LC50可以迅速有效地对农药的毒性进行初步判断,而生长抑制率和幼虫孵化数是更加敏感的评估农药对蚯蚓毒性的指标.     

International standardized procedures for in vivo evaluation of multi-walled carbon nanotube toxicity in water

The classical approach in ecotoxicological evaluation of chemical substances consists of conducting standardized bioassays on organism models. In this work, the potential impact of industrial multi-walled carbon nanotubes was investigated by ecotoxicological standardized procedures using aquatic organisms of different trophic levels, namely bacteria, green algae, invertebrates, fish, and amphibians. The results indicated (1) inhibition of growth in amphibians at 50 mg L^?1 and higher, and (2) no effects on daphnia and fish up to 100 mg L^?1. With the exception of algae (for which Fe deficiency is measured), it seems that the observed toxicity may be due to physiological effects in relation to the ingestion of carbon nanotubes not necessarily related to their intrinsic effects.     

A theoretical‐methodological approach towards the evaluation of environmental pollutants

The recognition and analysis as well as a generally applicable ecotoxicological evaluation are parts of the strategy towards the determination of changes in the quality of environmental compartments and their effects on humans. The progress in the field of identification of qualitative effects calls for the creation of a theoretical basis for the empirical findings. With this task being the main concern, fundamental ideas on a theoretical‐methodological approach towards the ecotoxicological evaluation of environmental chemicals are discussed. The relationships between the molecular structure and the effect of the substances in abiotic and biotic environmental compartments are the basis of this approach.     

A new ecotoxicological test method for genetically modified plants and other stressors in soil with the black fungus gnat <Emphasis Type="Italic">Bradysia impatiens</Emphasis> (Diptera): current status of test development and dietary effects of azadirachtin on larval development and emergence rate

Background

Few suitable and standardized test methods are currently available to test the effects of genetically modified plants (GMP) on non-target organisms. To fill this gap and improve ecotoxicological testing for GMP, we developed a new soil ecotoxicological test method using sciarid larvae as test organisms.

Results

Bradysia impatiens was identified as a candidate species. Species of the genus Bradysia occur in high numbers in European agroecosystems and B. impatiens can be reared in the laboratory in continuous culture. A functional basic test design was successfully developed. Newly hatched larvae were used as the initial life stage to cover most of the life cycle of the species during the test. Azadirachtin was identified as a suitable reference substance. In several tests, the effects of this substance on development time and emergence rate varied for different temperatures and test substrates. The toxicity was higher at 25 °C compared to 20 °C and in tropical artificial soil compared to coconut fiber substrate.

Conclusions and outlook

Results suggest that the developed test system is suitable to enter a full standardization process, e.g., via the Organisation for Economic Co-operation and Development. Such a standardization would not only assist the risk assessment of GMP, but could include other stressors such as systemic pesticides or veterinary pharmaceuticals reaching the soil, e.g., via spreading manure. The use of sciarid flies as test organisms supports recommendations of EFSA, which stressed the ecological role of flies and encouraged including Diptera into test batteries.

     

A multi-integrated approach on toxicity effects of engineered TiO2 nanoparticles

The new properties of engineered nanoparticles drive the need for new knowledge on the safety, fate, behavior and biologic effects of these particles on organisms and ecosystems. Titanium dioxide nanoparticles have been used extensively for a wide range of applications, e.g, self-cleaning surface coatings, solar cells, water treatment agents, topical sunscreens. Within this scenario increased environmental exposure can be expected but data on the ecotoxicological evaluation of nanoparticles are still scarce. The main purpose of this work was the evaluation of effects of TiO₂ nanoparticles in several organisms, covering different trophic levels, using a battery of aquatic assays. Using fish as a vertebrate model organism tissue histological and ultrastructural observations and the stress enzyme activity were also studied. TiO₂ nanoparticles (Aeroxide® P25), two phase composition of anatase (65%) and rutile (35%) with an average particle size value of 27.6±11 nm were used. Results on the EC₅₀ for the tested aquatic organisms showed toxicity for the bacteria, the algae and the crustacean, being the algae the most sensitive tested organism. The aquatic plant Lemna minor showed no effect on growth. The fish Carassius auratus showed no effect on a 21 day survival test, though at a biochemical level the cytosolic Glutathione-S-Transferase total activity, in intestines, showed a general significant decrease (p<0.05) after 14 days of exposure for all tested concentrations. The presence of TiO₂ nanoparticles aggregates were observed in the intestine lumen but their internalization by intestine cells could not be confirmed.