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.

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.

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.

Determining the agricultural ammonia immission using bark bio-monitoring: comparison with passive sampler measurements.

Bark samples of spruce, pine and oak trees were collected at two sites in southern Bavaria which are characterized by high agricultural ammonia emissions. The samples were taken using a recently developed bark sampling device which removes a defined layer of the bark. The bark was then analysed for ammonium concentration in order to reflect the environmental ammonia immission. The measured bark concentrations decreased with rising distance between the sample trees and the ammonia source. This applied (i) to measurements inside a closed forest stand ranging from forest edge with high immission to forest interior with much lower immission, and (ii) to the open field where single-standing trees were sampled. Comparing the ammonium concentrations among the three different tree species revealed significant correlations. Thus, it could be shown that old spruce trees are as usable for bark bio-monitoring as the traditionally used pine and oak trees. The ammonium concentrations of the bark were significantly correlated to measurements taken by ammonia passive samplers at the same locations. These results indicate that bark samples may be used for a standardised monitoring of airborne ammonia load. A major advantage of the technique is the determination of the long-term accumulative ammonia load using a single measurement.