Feasibility of assessing vegetative and generative endpoints of crop- and non- crop terrestrial plant species for non-target terrestrial plant (NTTP) regulatory testing under greenhouse conditions.

Agriculture is the dominating land-use in the EU member states covering nearly half of the surface area. Using herbicides to reduce weed competition in agricultural areas can adversely affect Non-Target Terrestrial Plants (NTTP) growing in field margins. According to the EFSA Scientific Opinion on NTTPs an important protection goal is to maintain the biodiversity of plant species in agricultural areas. EFSA recommends to include also non-crop species mentioned in OECD guidelines (OECD 208 and 227) in the testing and to assess not only vegetative but also generative endpoints during the plant life-cycle such as flowering and seed production. The objectives of this study were to evaluate the feasibility of assessing generative endpoints of crop and non-crop species for NTTP regulatory testing under greenhouse conditions and to assess if generative endpoints are more sensitive than vegetative endpoints. The experimental design consisted of one control and four herbicide (Atlantis® WG) application rates, with 6 replicates each. The application rates of the test substance were the maximum field rate and 30%, 10% and 3% of the field rate. Biomass, plant height, flowering, seed production as well as seedling emergence of the F1 generation were assessed. The study shows a feasible approach to assess vegetative and generative endpoints of (non-) crops species under greenhouse conditions on the basis of the OECD guideline 227. The vegetative endpoints plant height and biomass were not more sensitive if assessed during the generative growth stage when compared to the vegetative growth stage of the plants. In contrast to that, the generative endpoint seed production was partly more sensitive in comparison to the vegetative endpoints biomass and plant height. For regulatory NTTP studies, 5 or more test substance rates at non-lethal levels should be tested so to allow the determination of ER10/50 values for vegetative and generative endpoints.

Deriving criteria to select arthropod species for laboratory tests to assess the ecological risks from cultivating arthropod-resistant genetically engineered crops.

UNLABELLED: Arthropods form a major part of the biodiversity in agricultural landscapes. Many species are valued because they provide ecosystem services, including biological control, pollination and decomposition, or because they are of conservation interest. Some arthropods reduce crop yield and quality, and conventional chemical pesticides, biological control agents and genetically engineered (GE) crops are used to control them. A common concern addressed in the ecological risk assessment (ERA) that precedes regulatory approval of these pest control methods is their potential to adversely affect valued non-target arthropods (NTAs). A key concept of ERA is early-tier testing using worst-case exposure conditions in the laboratory and surrogate test species that are most likely to reveal an adverse effect. If no adverse effects are observed in those species at high exposures, confidence of negligible ecological risk from the use of the pest control method is increased. From experience with chemical pesticides and biological control agents, an approach is proposed for selecting test species for early-tier ERA of GE arthropod-resistant crops. Surrogate species should be selected that most closely meet three criteria: (i) Potential sensitivity: species should be the most likely to be sensitive to the arthropod-active compound based on the known spectrum of activity of the active ingredient, its mode of action, and the phylogenetic relatedness of the test and target species; (ii) RELEVANCE: species should be representative of valued taxa or functional groups that are most likely to be exposed to the arthropod-active compound in the field; and (iii) Availability and reliability: suitable life-stages of the test species must be obtainable in sufficient quantity and quality, and validated test protocols must be available that allow consistent detection of adverse effects on ecologically relevant parameters. Our proposed approach ensures that the most suitable species are selected for testing and that the resulting data provide the most rigorous test of the risk hypothesis of no adverse effect in order to increase the quality and efficiency of ERAs for cultivation of GE crops.

Recommendations for the design of laboratory studies on non-target arthropods for risk assessment of genetically engineered plants.

This paper provides recommendations on experimental design for early-tier laboratory studies used in risk assessments to evaluate potential adverse impacts of arthropod-resistant genetically engineered (GE) plants on non-target arthropods (NTAs). While we rely heavily on the currently used proteins from Bacillus thuringiensis (Bt) in this discussion, the concepts apply to other arthropod-active proteins. A risk may exist if the newly acquired trait of the GE plant has adverse effects on NTAs when they are exposed to the arthropod-active protein. Typically, the risk assessment follows a tiered approach that starts with laboratory studies under worst-case exposure conditions; such studies have a high ability to detect adverse effects on non-target species. Clear guidance on how such data are produced in laboratory studies assists the product developers and risk assessors. The studies should be reproducible and test clearly defined risk hypotheses. These properties contribute to the robustness of, and confidence in, environmental risk assessments for GE plants. Data from NTA studies, collected during the analysis phase of an environmental risk assessment, are critical to the outcome of the assessment and ultimately the decision taken by regulatory authorities on the release of a GE plant. Confidence in the results of early-tier laboratory studies is a precondition for the acceptance of data across regulatory jurisdictions and should encourage agencies to share useful information and thus avoid redundant testing.

Assessment of risk of insect-resistant transgenic crops to nontarget arthropods.

An international initiative is developing a scientifically rigorous approach to evaluate the potential risks to nontarget arthropods (NTAs) posed by insect-resistant, genetically modified (IRGM) crops. It adapts the tiered approach to risk assessment that is used internationally within regulatory toxicology and environmental sciences. The approach focuses on the formulation and testing of clearly stated risk hypotheses, making maximum use of available data and using formal decision guidelines to progress between testing stages (or tiers). It is intended to provide guidance to regulatory agencies that are currently developing their own NTA risk assessment guidelines for IRGM crops and to help harmonize regulatory requirements between different countries and different regions of the world.