Pressure from insect-resistant maize on protected butterflies and moths.

Intensification in agriculture affects many insect species, including butterflies. Insect-resistant crops, such as Bt (Bacillus thuringiensis) maize, which produces a toxin active against Lepidoptera, are an alternative to insecticide sprays. Genetically modified crops are regulated in most countries and require an environmental risk assessment. In the European Union, such assessments include the use of simulation models to predict the effects on nontarget Lepidoptera (NTL). To support the assessment of protected NTL, we extended an individual-based, stochastic, spatially explicit mathematical model (LepiX) to include a wider range of exposure scenarios, a species-sensitivity distribution, and an option for repeated exposure of individuals. We applied the model to transgenic maize DAS-1507, which expresses a high concentration of Bt toxin in pollen that may be consumed by NTL larvae on their host plants nearby. Even in the most conservative scenario without repeated exposure, mortality estimates for highly sensitive species ranged from 41% to 6% at distances of 10-1000 m from the nearest maize field. Repeated exposure can cause additional mortality and thus is relevant for the overall risk assessment. Uncertainties in both exposure and ecotoxicity estimates strongly influenced the predicted mortalities. Care should be taken to include these uncertainties in the model scenarios used for decision-making. In accordance with other modeling results, our simulations demonstrated that mean mortality may not be safe for protected species. With its high pollen expression, DAS-1507 maize may pose risks to sensitive and protected butterfly and moth species that may be difficult to manage. High expression of Bt toxin in pollen is unnecessary for controlling target pests. Consequently, we suggest that Bt maize with high pollen expression not be cultivated in regions where protected butterflies are to be conserved.

La intensificación en la agricultura afecta a muchas especies de insectos, incluyendo a las mariposas. Los cultivos resistentes a los insectos, como el maíz Bt (Bacillus thuringiensis), el cual produce una toxina activa contra los lepidópteros, son una alternativa a los insecticidas. Los cultivos genéticamente modificados (GM) están regulados en la mayoría de los países y requieren de una evaluación de riesgo ambiental. En la Unión Europea (EU), dichas evaluaciones incluyen el uso de modelos de simulación para pronosticar los efectos sobre los lepidópteros no objetivo (LNO). Para apoyar a la evaluación de LNO protegidos, extendimos un modelo matemático espacialmente explícito, estocástico y basado en el individuo (LepiX) para incluir una mayor gama de escenarios de exposición, una distribución de la sensibilidad de las especies y una opción para la exposición repetida de los individuos. Aplicamos el modelo al maíz transgénico DAS­1507, el cual expresa una alta concentración de toxina Bt en el polen que puede ser consumido por las larvas de LNO en una planta hospedera cercana. Incluso en el escenario más conservador sin una exposición repetida, las estimaciones de mortalidad para las especies altamente sensibles variaron entre el 41% y el 6% en distancias de 10­1000 m a partir del campo de maíz más cercano. La exposición repetida puede causar mortalidad adicional y por lo tanto es relevante para la evaluación general del riesgo. La incertidumbre en las estimaciones de la exposición y la ecotoxicidad influyeron fuertemente sobre la mortalidad pronosticada. Se debe tener cuidado de incluir estas incertidumbres en los escenarios modelados usados para la toma de decisiones. De acuerdo con los resultados de otros modelos, nuestras simulaciones demostraron que la mortalidad media podría no ser segura para las especies protegidas. Con su alta producción de polen, el maíz DAS­1507 podría representar un riesgo difícil de manejar para las especies de mariposas y polillas sensibles y protegidas. No se necesita una expresión elevada de la toxina Bt en el polen para controlar a las plagas. En consecuencia, sugerimos que no se cultive el maíz Bt con una alta producción de polen en las regiones en donde se busca conservar a las mariposas protegidas. Presión del maíz resistente a insectos sobre mariposas y polillas protegidas.

Assessing Effects of Genetically Modified Plant Material on the Aquatic Environment Using higher-tier Studies.

Genetically modified organisms are used extensively in agriculture. To assess potential side effects of genetically modified (GM) plant material on aquatic ecosystems, only a very small number of higher-tier studies have been performed. At the same time, these studies are particularly important for comprehensive risk assessment covering complex ecological relationships. Here we evaluate the methods of experimental higher-tier effect studies with GM plant material (or Bt toxin) in comparison to those well-established for pesticides. A major difference is that nominal test concentrations and thus dose-response relationships cannot easily be produced with GM plant material. Another important difference, particularly to non-systemic pesticides, is that aquatic organisms are exposed to GM plant material primarily through their feed. These and further differences in test requirements, compared with pesticides, call for a standardisation for GM-specific higher-tier study designs to assess their potentially complex effects in the aquatic ecosystems comprehensively.

The Effect of Mating Complexity on Gene Drive Dynamics.

AbstractGene drive technology promises to deliver on some of the global challenges humanity faces today in health care, agriculture, and conservation. However, there is a limited understanding of the consequences of releasing self-perpetuating transgenic organisms into wild populations under complex ecological conditions. In this study, we analyze the impact of three such complexities-mate choice, mating systems, and spatial mating network-on the population dynamics for two distinct classes of modification gene drive systems. All three factors had a high impact on the modeling outcome. First, we demonstrate that distortion-based gene drives appear to be more robust against mate choice than viability-based gene drives. Second, we find that gene drive spread is much faster for higher degrees of polygamy. Including a fitness cost, the drive is fastest for intermediate levels of polygamy. Finally, the spread of a gene drive is faster and more effective when the individuals have fewer connections in a spatial mating network. Our results highlight the need to include mating complexities when modeling the properties of gene drives, such as release thresholds, timescales, and population-level consequences. This inclusion will enable a more confident prediction of the dynamics of engineered gene drives and possibly even inform about the origin and evolution of natural gene drives.

Effect of Bt toxin Cry1Ab on two freshwater caddisfly shredders - an attempt to establish dose-effect relationships through food-spiking.

Genetically modified organisms (GMOs), which produce Bacillus thuringiensis (Bt) toxins, are widely used in agriculture in some parts of the world. Despite this, ecotoxicological methods, tailored to GMOs, are lacking to assess effects on aquatic environments. With the objective to investigate a food-related exposure pathway for aquatic shredders, we used a new food-spiking method while caddisfly larvae (Chaetopteryx spec., Sericostoma spec.) served as test species. Pure Cry1Ab toxins were spiked on black alder leaf discs and subsequently used in a feeding experiment. The toxin did not influence larval mortality compared to the control. The results, however, showed significant effects on larval lipid content (Chaetopteryx spec.) and development (Sericostoma spec.) at concentrations of 17.2 and 132.4 ng Cry1Ab/mg leaf, respectively. These changes are indicative for impacts on the fitness of the specimen and thus relevant in a risk assessment context. Ultimately, the food-spiking method allowed applying different Bt toxin concentrations leading to the establishment of dose-response relationships for various response variables. The use of long test durations and sublethal endpoints (consumption, lipid content, growth, larval instars) is, moreover, advisable when testing GMO effects.

A new ecotoxicological test method for genetically modified plants and other stressors in soil with the black fungus gnat Bradysia impatiens (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.

Impact of genetically modified organisms on aquatic environments: Review of available data for the risk assessment.

The aquatic environment is strongly connected to the surrounding agricultural landscapes, which regularly serve as sources of stressors such as agrochemicals. Genetically modified crops, which are cultivated on a large scale in many countries, may also act as stressors. Despite the commercial use of genetically modified organisms (GMOs) for over 20years, their impact on the aquatic environment came into focus only 10years ago. We present the status quo of the available scientific data in order to provide an input for informed aquatic risk assessment of GMOs. We could identify only 39 publications, including 84 studies, dealing with GMOs in the aquatic environment, and our analysis shows substantial knowledge gaps. The available information is restricted to a small number of crop plants, traits, events, and test organisms. The analysis of effect studies reveals that only a narrow range of organisms has been tested and that studies on combinatorial actions of stressors are virtually absent. The analysis of fate studies shows that many aspects, such as the fate of leached toxins, degradation of plant material, and distribution of crop residues in the aquatic habitat, are insufficiently investigated. Together with these research needs, we identify standardization of test methods as an issue of high priority, both for research and risk assessment needed for GMO regulation.

Procedure to select test organisms for environmental risk assessment of genetically modified crops in aquatic systems.

For a long time, the environmental risk assessment (ERA) of genetically modified (GM) crops focused mainly on terrestrial ecosystems. This changed when it was scientifically established that aquatic ecosystems are exposed to GM crop residues that may negatively affect aquatic species. To assist the risk assessment process, we present a tool to identify ecologically relevant species usable in tiered testing prior to authorization or for biological monitoring in the field. The tool is derived from a selection procedure for terrestrial ecosystems with substantial but necessary changes to adequately consider the differences in the type of ecosystems. By using available information from the Water Framework Directive (2000/60/EC), the procedure can draw upon existing biological data on aquatic systems. The proposed procedure for aquatic ecosystems was tested for the first time during an expert workshop in 2013, using the cultivation of Bacillus thuringiensis (Bt) maize as the GM crop and 1 stream type as the receiving environment in the model system. During this workshop, species executing important ecological functions in aquatic environments were identified in a stepwise procedure according to predefined ecological criteria. By doing so, we demonstrated that the procedure is practicable with regard to its goal: From the initial long list of 141 potentially exposed aquatic species, 7 species and 1 genus were identified as the most suitable candidates for nontarget testing programs. Integr Environ Assess Manag 2017;13:974-979. © 2017 SETAC.

Pediatric nasolacrimal duct obstruction-benefit of a combined therapeutic approach.

BACKGROUND: Pediatric nasolacrimal duct obstruction (PNDO) requires therapeutic intervention after conservative procedures failed. As resilient treatment guidelines for the treatment are missing, the aim of this study was to evaluate the advantages of two different intervention techniques in children with PNDO. METHODS: Between January, 2006 and June, 2014, 233 children (0-208 months) were treated either with conventional probing by ophthalmologists only (Group I) or with endonasal endoscopic interdisciplinary approach (Group II). The clinical outcome was analyzed. RESULTS: The overall success rate of Group I was 93.4% compared to 98.4% of Group II (P<0.05). 50% of all interventions (n=62) of Group II required further surgical procedures in addition to probing/irrigation, particularly with regard to children <6 and >24 months. CONCLUSIONS: Endoscopic control in treatment of PNDO allows exact identification of the stenosis and appropriate surgical intervention with an improved clinical outcome. Endonasal endoscopic surgical techniques should be the standard PNDO treatment.

Reply to the EFSA (2016) on the relevance of recent publications (Hofmann et al. 2014, 2016) on environmental risk assessment and management of Bt-maize events (MON810, Bt11 and 1507).

In this commentary, we respond to a report of the EFSA GMO Panel (EFSA EFSA Supp Publ, 1) that criticises the outcomes of two studies published in this journal (Hofmann et al. Environ Sci Eur 26: 24, 2; Environ Sci Eur 28: 14, 3). Both publications relate to the environmental risk assessment and management of Bt-maize, including maize events MON810, Bt11 and maize 1507. The results of Hofmann et al. (Environ Sci Eur 26: 24, 2), using standardised pollen mass filter deposition measurements, indicated that the EFSA Panel model had underestimated pollen deposition and, hence, exposure of non-target organisms to Bt-maize pollen. The results implied a need for safety buffer distances in the kilometre range for protected nature reserve areas instead of the 20-30 m range recommended by the EFSA Panel. As a result, the EFSA Panel revised their model (EFSA EFSA J 13: 4127, 4), adopting the slope of the empirical data from Hofmann et al. The intercept, however, was substantially reduced to less than 1% at one point by introducing further assumptions based on the estimates of mainly panel members, citing possible 'uncertainty'. Hofmann et al. (Environ Sci Eur 28: 14, 3) published extensive empirical data regarding pollen deposition on leaves. These results were part of a larger 3-year study involving detailed measurements of pollen release, dispersal and deposition over the maize flowering period. The data collected in situ confirmed the previous predictions of Hofmann et al. (Environ Sci Eur 26: 24, 2). Mean levels and observed variability of pollen deposition on maize and four lepidopteran host plants exceeded the assumptions and disagreed with the conclusions of the EFSA Panel. The EFSA Panel reacted in a report (EFSA EFSA Supp Publ, 1) criticising the methods and outcomes of the two published studies of Hofmann et al. while reaffirming their original recommendations. We respond here point-by-point, showing that the critique is not justified. Based on our results on Urtica leaf pollen density, we confirm the need for specific environmental impact assessments for Bt-maize cultivation with respect to protected habitats within isolation buffer distances in the kilometre range.

Erratum to: Reply to the EFSA (2016) on the relevance of recent publications (Hofmann et al. 2014, 2016) on environmental risk assessment and management of Bt-maize events (MON810, Bt11 and 1507).

[This corrects the article DOI: 10.1186/s12302-017-0106-0.].

Accumulation and variability of maize pollen deposition on leaves of European Lepidoptera host plants and relation to release rates and deposition determined by standardised technical sampling.

BACKGROUND: Risk assessment for GMOs such as Bt maize requires detailed data concerning pollen deposition onto non-target host-plant leaves. A field study of pollen on lepidopteran host-plant leaves was therefore undertaken in 2009-2012 in Germany. During the maize flowering period, we used in situ microscopy at a spatial resolution adequate to monitor the feeding behaviour of butterfly larvae. The plant-specific pollen deposition data were supplemented with standardised measurements of pollen release rates and deposition obtained by volumetric pollen monitors and passive samplers. RESULTS: In 2010, we made 5377 measurements of maize pollen deposited onto leaves of maize, nettle, goosefoot, sorrel and blackberry. Overall mean leaf deposition during the flowering period ranged from 54 to 478 n/cm2 (grains/cm2) depending on plant species and site, while daily mean leaf deposition values were as high as 2710 n/cm2. Maximum single leaf-deposition values reached up to 103,000 n/cm2, with a 95 % confidence-limit upper boundary of 11,716 n/cm2. CONCLUSIONS: Daily means and variation of single values uncovered by our detailed measurements are considerably higher than previously assumed. The recorded levels are more than a single degree of magnitude larger than actual EU expert risk assessment assumptions. Because variation and total aggregation of deposited pollen on leaves have been previously underestimated, lepidopteran larvae have actually been subjected to higher and more variable exposure. Higher risks to these organisms must consequently be assumed. Our results imply that risk assessments related to the effects of Bt maize exposure under both realistic cultivation conditions and worst-case scenarios must be revised. Under common cultivation conditions, isolation buffer distances in the kilometre range are recommended rather than the 20-30 m distance defined by the EFSA.

Prioritizing stream types according to their potential risk to receive crop plant material--A GIS-based procedure to assist in the risk assessment of genetically modified crops and systemic insecticide residues.

Crop plant residues may enter aquatic ecosystems via wind deposition or surface runoff. In the case of genetically modified crops or crops treated with systemic pesticides, these materials may contain insecticidal Bt toxins or pesticides that potentially affect aquatic life. However, the particular exposure pattern of aquatic ecosystems (i.e., via plant material) is not properly reflected in current risk assessment schemes, which primarily focus on waterborne toxicity and not on plant material as the route of uptake. To assist in risk assessment, the present study proposes a prioritization procedure of stream types based on the freshwater network and crop-specific cultivation data using maize in Germany as a model system. To identify stream types with a high probability of receiving crop materials, we developed a formalized, criteria-based and thus transparent procedure that considers the exposure-related parameters, ecological status--an estimate of the diversity and potential vulnerability of local communities towards anthropogenic stress--and availability of uncontaminated reference sections. By applying the procedure to maize, ten stream types out of 38 are expected to be the most relevant if the ecological effects from plant-incorporated pesticides need to be evaluated. This information is an important first step to identifying habitats within these stream types with a high probability of receiving crop plant material at a more local scale, including accumulation areas. Moreover, the prioritization procedure developed in the present study may support the selection of aquatic species for ecotoxicological testing based on their probability of occurrence in stream types having a higher chance of exposure. Finally, this procedure can be adapted to any geographical region or crop of interest and is, therefore, a valuable tool for a site-specific risk assessment of crop plants carrying systemic pesticides or novel proteins, such as insecticidal Bt toxins, expressed in genetically modified crops.

Feeding Behaviour on Host Plants May Influence Potential Exposure to Bt Maize Pollen of Aglais Urticae Larvae (Lepidoptera, Nymphalidae).

Non-target butterfly larvae may be harmed by feeding on host plants dusted with Bt maize pollen. Feeding patterns of larvae and their utilization of host plants can affect the adverse Bt impact because the maize pollen is distributed unequally on the plant. In a field study, we investigated the feeding of larvae of the Small Tortoiseshell, Aglais urticae, on nettles, Urtica dioica. Young larvae used smaller host plants than older larvae. In general, the position of the larvae was in the top part of the host plant, but older larvae showed a broader vertical distribution on the nettles. Leaf blades and leaf tips were the plant parts most often consumed. Leaf veins were consumed but midribs were fed on to a lesser extent than other plant veins, particularly by young larvae. The feeding behavior of the larvae may increase possible exposure to Bt maize pollen because pollen densities are expected to be higher on the top parts and along leaf veins of nettles.

Endonasal nasolacrimal duct surgery: a comparative study of two techniques.

Dacryocystorhinostomy (DCR) is performed in patients with saccal or postsaccal lacrimal duct obstruction. Focusing on the endonasal approach, we compared success rates, clinical outcome, complications and patient satisfaction of endoscopic vs. non-endoscopic techniques in endonasal DCR. We analyzed the results of 173 patients who underwent an endonasal DCR either utilizing a non-endoscopic (Group I) or an endoscopic technique (Group II) between 2006 and 2011. Irrigation tests followed the first day and at least 3 months after surgery. Postoperative patients' satisfaction and the occurrence of symptoms were documented and evaluated in a follow-up questionnaire. The minor complication rates of both endonasal DCR techniques were similarly low (10%) without severe adverse events. The use of the endoscope prolonged the operating time significantly (28 ± 9 min Group I vs. 34 ± 14 min Group II, p = 0.003). The success rate was 90.2% in Group II compared to only 57.9% in Group I (p < 0.000). Further, we determined the absence of reflux during the irrigation test 1 day after surgery as a significant predictor for the later outcome (R = 1.598, p = 0.005). The follow-up questionnaire revealed a significant improvement in subjectively perceived symptoms by the surgical intervention for both endonasal techniques (p < 0.000). The endoscopically assisted DCR is a safe and successful endonasal technique for patients with saccal or postsaccal lacrimal duct obstructions. The use of the endoscope led to significant higher success rates compared to non-endoscopic techniques in our collective.

2011 IEEE Visualization Contest winner: Visualizing unsteady vortical behavior of a centrifugal pump.

In the 2011 IEEE Visualization Contest, the dataset represented a high-resolution simulation of a centrifugal pump operating below optimal speed. The goal was to find suitable visualization techniques to identify regions of rotating stall that impede the pump's effectiveness. The winning entry split analysis of the pump into three parts based on the pump's functional behavior. It then applied local and integration-based methods to communicate the unsteady flow behavior in different regions of the dataset. This research formed the basis for a comparison of common vortex extractors and more recent methods. In particular, integration-based methods (separation measures, accumulated scalar fields, particle path lines, and advection textures) are well suited to capture the complex time-dependent flow behavior. This video (http://youtu.be/oD7QuabY0oU) shows simulations of unsteady flow in a centrifugal pump.

A New Method for in Situ Measurement of Bt-Maize Pollen Deposition on Host-Plant Leaves.

Maize is wind pollinated and produces huge amounts of pollen. In consequence, the Cry toxins expressed in the pollen of Bt maize will be dispersed by wind in the surrounding vegetation leading to exposure of non-target organisms (NTO). NTO like lepidopteran larvae may be affected by the uptake of Bt-pollen deposited on their host plants. Although some information is available to estimate pollen deposition on host plants, recorded data are based on indirect measurements such as shaking or washing off pollen, or removing pollen with adhesive tapes. These methods often lack precision and they do not include the necessary information such as the spatial and temporal variation of pollen deposition on the leaves. Here, we present a new method for recording in situ the amount and the distribution of Bt-maize pollen deposited on host plant leaves. The method is based on the use of a mobile digital microscope (Dino-Lite Pro, including DinoCapture software), which can be used in combination with a notebook in the field. The method was evaluated during experiments in 2008 to 2010. Maize pollen could be correctly identified and pollen deposition as well as the spatial heterogeneity of maize pollen deposition was recorded on maize and different lepidopteran host plants (Centaurea scabiosa, Chenopodium album, Rumex spp., Succina pratensis and Urtica dioica) growing adjacent to maize fields.

First asymmetric synthesis of (-)-lintetralin via intramolecular Friedel-Crafts-type cyclization.

The asymmetric synthesis of an aryltetralin lignan, (-)-lintetralin, was achieved with an overall yield of 29% with seven steps. Key features of the synthesis are an asymmetric Strecker reaction, a diastereoselective Michael addition of the lithiated amino nitrile product to 5H-furan-2-one, and an intramolecular carbocationic cyclization to provide the desired ring skeleton with the correct configuration.

The developmental strategy of an idiobiont ectoparasitoid, Dendrocerus carpenteri : influence of variations in host quality on offspring growth and fitness.

Dendrocerus carpenteri (Curtis) (Hymenoptera: Megaspilidae) is a solitary hyperparasitoid, which attacks prepupal and pupal stages of hymenopteran parasitoids inside mummified aphids. The larva feeds externally on the host, which is envenomed by the female at oviposition. To evaluate the influence of variations in host quality on the growth, development and fitness of D. carpenteri, we varied the size and developmental stage of the primary parasitoid host (Aphidius ervi Haliday), which was reared on different instars of pea aphid [Acyrthosiphon pisum (Harris)] and English grain aphid [Sitobion avenae (F.)]. Within each kind of host, females eclosed from the relatively larger mummies, while males eclosed from the smaller mummies. Host size and hyperparasitoid size were correlated, and females were larger than males. In hyperparasitoids developing on prepupal and pupal hosts, development time from oviposition to adult eclosion was proportional to size; females required more time for development than males. The mean relative growth rate was the same in males and females and increased with host quality, as predicted by the growth model of Mackauer and Sequeira (1993) for idiobiont parasitoids. Larvae developing on late-pupal stages and pharate adults of A. ervi were unable to consume sclerotized host tissues; they were smaller and needed more time for development. The average number of mature eggs at eclosion was six, except in females developing on suboptimal hosts, which contained only one egg or none. Egg volume was correlated with female size, possibly reflecting differences in larval ontogeny. We provide equations describing the relationship between host quality as indexed by hind-tibia length of the mummified aphid and adult body size in terms of dry mass, development time and mean relative growth rate of D. carpenteri. We discuss the usefulness of host size as a proxy of host quality for idiobiont parasitoids, and provide examples of exceptions.