A laboratory soil degradation study was conducted to determine the biodegradation potential of a DvSnf7 dsRNA transcript derived from a Monsanto genetically modified (GM) maize product that confers resistance to corn rootworm (CRW; Diabrotica spp.). This study provides new information to improve the environmental assessment of dsRNAs that become pesticidal through an RNAi process. Three agricultural soils differing in their physicochemical characteristics were obtained from the U.S., Illinois (IL; silt loam), Missouri (MO; loamy sand) and North Dakota (ND; clay loam), and exposed to the target dsRNA by incorporating insect-protected maize biomass and purified (in vitro-transcribed) DvSnf7 RNA into soil. The GM and control (non-GM maize) materials were added to each soil and incubated at ca. 22°C for 48 hours (h). Samples were collected at 12 time intervals during the incubation period, extracted, and analyzed using QuantiGene molecular analysis and insect bioassay methods. The DT50 (half-life) values for DvSnf7 RNA in IL, MO, and ND soils were 19, 28, and 15 h based on QuantiGene, and 18, 29, and 14 h based on insect bioassay, respectively. Furthermore, the DT90 (time to 90% degradation) values for DvSnf7 RNA in all three soils were <35 h. These results indicate that DvSnf7 RNA was degraded and biological activity was undetectable within approximately 2 days after application to soil, regardless of texture, pH, clay content and other soil differences. Furthermore, soil-incorporated DvSnf7 RNA was non-detectable in soil after 48 h, as measured by QuantiGene, at levels ranging more than two orders of magnitude (0.3, 1.5, 7.5 and 37.5 µg RNA/g soil). Results from this study indicate that the DvSnf7 dsRNA is unlikely to persist or accumulate in the environment. Furthermore, the rapid degradation of DvSnf7 dsRNA provides a basis to define relevant exposure scenarios for future RNA-based agricultural products.

As part of an environmental risk assessment, the potential impact of genetically modified (GM) maize MON 87411 on non-target arthropods (NTAs) was evaluated in the field. MON 87411 confers resistance to corn rootworm (CRW; Diabrotica spp.) by expressing an insecticidal double-stranded RNA (dsRNA) transcript and the Cry3Bb1 protein and tolerance to the herbicide glyphosate by producing the CP4 EPSPS protein. Field trials were conducted at 14 sites providing high geographic and environmental diversity within maize production areas from three geographic regions including the U.S., Argentina, and Brazil. MON 87411, the conventional control, and four commercial conventional reference hybrids were evaluated for NTA abundance and damage. Twenty arthropod taxa met minimum abundance criteria for valid statistical analysis. Nine of these taxa occurred in at least two of the three regions and in at least four sites across regions. These nine taxa included: aphid, predatory earwig, lacewing, ladybird beetle, leafhopper, minute pirate bug, parasitic wasp, sap beetle, and spider. In addition to wide regional distribution, these taxa encompass the ecological functions of herbivores, predators and parasitoids in maize agro-ecosystems. Thus, the nine arthropods may serve as representative taxa of maize agro-ecosystems, and thereby support that analysis of relevant data generated in one region can be transportable for the risk assessment of the same or similar GM crop products in another region. Across the 20 taxa analyzed, no statistically significant differences in abundance were detected between MON 87411 and the conventional control for 123 of the 128 individual-site comparisons (96.1 %). For the nine widely distributed taxa, no statistically significant differences in abundance were detected between MON 87411 and the conventional control. Furthermore, no statistically significant differences were detected between MON 87411 and the conventional control for 53 out of 56 individual-site comparisons (94.6 %) of NTA pest damage to the crop. In each case where a significant difference was observed in arthropod abundance or damage, the mean value for MON 87411 was within the reference range and/or the difference was not consistently observed across collection methods and/or sites. Thus, the differences were not representative of an adverse effect unfamiliar to maize and/or were not indicative of a consistent plant response associated with the GM traits. Results from this study support a conclusion of no adverse environmental impact of MON 87411 on NTAs compared to conventional maize and demonstrate the utility of relevant transportable data across regions for the ERA of GM crops.Electronic supplementary materialThe online version of this article (doi:10.1007/s11248-015-9907-3) contains supplementary material, which is available to authorized users.

As part of an ecological risk assessment, Roundup Ready 2 Yield® soybean (MON 89788) was compared to a conventional control soybean variety, A3244, for disease and arthropod damage, plant response to abiotic stress and cold, effects on succeeding plant growth (allelopathic effects), plant response to a bacterial symbiont, and effects on the ability of seed to survive and volunteer in a subsequent growing season. Statistically significant differences between MON 89788 and A3244 were considered in the context of the genetic variation known to occur in soybean and were assessed for their potential impact on plant pest (weed) potential and adverse environmental impact. The results of these studies revealed no effects of the genetic modification that would result in increased pest potential or adverse environmental impact of MON 89788 compared with A3244. This paper illustrates how such characterization studies conducted in a range of environments where the crop is grown are used in an ecological risk assessment of the genetically modified (GM) crop. Furthermore, risk assessors and decision makers use this information when deciding whether to approve a GM crop for cultivation in-or grain import into-their country.

The insidious flower bug, Orius insidiosus (Say) (Heteroptera: Anthocoridae) is an important surrogate species for assessing potential effects of plant-incorporated protectants (PIPs) on nontarget heterotrophic predators. In this study, a continuous dietary exposure system was optimized by assessing the effect of diet composition and age on the survival and development of nymphs of O. insidiosus. Greater than 85% control survival and an acceptable rate of development from nymph hatching to adult was achieved using 5-d-old nymphs at test initiation and a bee pollen-based diet supplemented with 25% Ephestia eggs. There was an unacceptable level of mortality (>40%) and/or a significantly prolonged development time when nymphs were <5 d old at test initiation. When 5-d-old nymphs were fed a bee pollen diet containing 25% Ephestia eggs and 100 μg/g potassium arsenate, time-dependent mortality was observed with a median lethal time (LT50) of 4.4 d and 100% mortality was observed after 10 d of feeding, indicating the effectiveness of the test system to detect adverse effects by dietary exposure. It is recommended that well-defined 5-d-old nymphs and an encapsulated bee pollen-based diet containing 25% ground Ephestia eggs be used in a Tier-I dietary feeding exposure assay for detecting potential effects of PIPs on O. insidiosus nymphs.