Mark K. Sears

Impact of Bt corn pollen on monarch butterfly populations: A risk assessment

A collaborative research effort by scientists in several states and in Canada has produced information to develop a formal risk assessment of the impact of Bt corn on monarch butterfly (Danaus plexippus) populations. Information was sought on the acute toxic effects of Bt corn pollen and the degree to which monarch larvae would be exposed to toxic amounts of Bt pollen on its host plant, the common milkweed, Asclepias syriaca, found in and around cornfields. Expression of Cry proteins, the active toxicant found in Bt corn tissues, differed among hybrids, and especially so in the concentrations found in pollen of different events. In most commercial hybrids, Bt expression in pollen is low, and laboratory and field studies show no acute toxic effects at any pollen density that would be encountered in the field. Other factors mitigating exposure of larvae include the variable and limited overlap between pollen shed and larval activity periods, the fact that only a portion of the monarch population utilizes milkweed stands in and near cornfields, and the current adoption rate of Bt corn at 19% of North American corn-growing areas. This 2-year study suggests that the impact of Bt corn pollen from current commercial hybrids on monarch butterfly populations is negligible.

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.

Monarch larvae sensitivity to Bacillus thuringiensis- purified proteins and pollen

Laboratory tests were conducted to establish the relative toxicity of Bacillus thuringiensis (Bt) toxins and pollen from Bt corn to monarch larvae. Toxins tested included Cry1Ab, Cry1Ac, Cry9C, and Cry1F. Three methods were used: (i) purified toxins incorporated into artificial diet, (ii) pollen collected from Bt corn hybrids applied directly to milkweed leaf discs, and (iii) Bt pollen contaminated with corn tassel material applied directly to milkweed leaf discs. Bioassays of purified Bt toxins indicate that Cry9C and Cry1F proteins are relatively nontoxic to monarch first instars, whereas first instars are sensitive to Cry1Ab and Cry1Ac proteins. Older instars were 12 to 23 times less susceptible to Cry1Ab toxin compared with first instars. Pollen bioassays suggest that pollen contaminants, an artifact of pollen processing, can dramatically influence larval survival and weight gains and produce spurious results. The only transgenic corn pollen that consistently affected monarch larvae was from Cry1Ab event 176 hybrids, currently <2% corn planted and for which re-registration has not been applied. Results from the other types of Bt corn suggest that pollen from the Cry1Ab (events Bt11 and Mon810) and Cry1F, and experimental Cry9C hybrids, will have no acute effects on monarch butterfly larvae in field settings.

Corn pollen deposition on milkweeds in and near cornfields

The density of corn pollen on leaves of milkweed plants inside and outside of cornfields was measured in several studies from different localities. The purpose was to obtain a representative picture of naturally occurring pollen densities to provide a perspective for laboratory and field studies of monarch larvae feeding on milkweed leaves with Bt corn pollen. Pollen density was highest (average 170.6 grains per cm2) inside the cornfield and was progressively lower from the field edge outward, falling to 14.2 grains per cm2 at 2 m. Inside the cornfield, and for each distance from the field edge, a frequency distribution is presented showing the proportion of leaf samples with different pollen densities. Inside cornfields, 95% of leaf samples had pollen densities below 600 grains per cm2 and the highest pollen density observed was 1400 grains per cm2, which occurred in a study with a rainless anthesis period. All other studies had rainfall events during the anthesis period. A single rain event can remove 54–86% of the pollen on leaves. Leaves on the upper portion of milkweed plants, where young monarch larvae tend to feed, had only 30–50% of the pollen density levels of middle leaves.

Temporal and spatial overlap between monarch larvae and corn pollen

To assess the likelihood that monarch larvae will be exposed to Bacillus thuringiensis (Bt) pollen, we studied milkweed and monarch densities in habitats which comprise much of the land available to breeding monarchs, e.g., cornfields, cornfield edges, other agricultural fields, and nonagricultural areas, in four regions of the monarch breeding range. We found that monarchs use milkweed in cornfields throughout their breeding season, and that per plant densities are as high or higher in agricultural habitats as in nonagricultural habitats. As a result of the prevalence of agricultural land, most of the monarchs produced in the upper Midwest are likely to originate in cornfields or other agricultural habitats. There was a greater temporal overlap between susceptible monarchs and corn anthesis in the northern than the southern part of the summer breeding range, because of earlier pollen shed in the south. The importance of agricultural habitats to monarch production suggests that, regardless of the impact of genetically modified crops, agricultural practices such as weed control and foliar insecticide use could have large impacts on monarch populations.

Assessing the impact of Cry1Ab-expressing corn pollen on monarch butterfly larvae in field studies

Survival and growth of monarch larvae, Danaus plexippus (L.), after exposure to either Cry1Ab-expressing pollen from three Bacillus thuringiensis (Bt) corn (Zea mays L.) events differing in toxin expression or to the insecticide, λ-cyhalothrin, were examined in field studies. First instars exposed to low doses (≈22 grains per cm2) of event-176 pollen gained 18% less weight than those exposed to Bt11 or Mon810 pollen after a 5-day exposure period. Larvae exposed to 67 pollen grains per cm2 on milkweed leaves from within an event-176 field exhibited 60% lower survivorship and 42% less weight gain compared with those exposed to leaves from outside the field. In contrast, Bt11 pollen had no effect on growth to adulthood or survival of first or third instars exposed for 5 days to ≈55 and 97 pollen grains per cm2, respectively. Similarly, no differences in larval survivorship were observed after a 4-day exposure period to leaves with 504–586 (within fields) or 18–22 (outside the field) pollen grains per cm2 collected from Bt11 and non-Bt sweet-corn fields. However, survivorship and weight gain were drastically reduced in non-Bt fields treated with λ-cyhalothrin. The effects of Bt11 and Mon810 pollen on the survivorship of larvae feeding 14 to 22 days on milkweeds in fields were negligible. Further studies should examine the lifetime and reproductive impact of Bt11 and Mon810 pollen on monarchs after long-term exposure to naturally deposited pollen.

Effects on Monarch Butterfly Larvae (Lepidoptera: Danaidae) After Continuous Exposure to Cry1Ab-Expressing Corn During Anthesis

Abstract Effects on monarch butterfly, Danaus plexippus L., after continuous exposure of larvae to natural deposits of Bacillus thuringiensis (Bt) and non-Bt pollen on milkweed, were measured in five studies. First instars were exposed at 3–4 and 6–7 d after initial anthesis, either directly on milkweed plants in commercial cornfields or in the laboratory on leaves collected from milkweeds in corn plots. Pollen exposure levels ranging from 122 to 188 grains/cm2/d were similar to within-field levels that monarch butterfly populations might experience in the general population of cornfields. Results indicate that 23.7% fewer larvae exposed to these levels of Bt pollen during anthesis reached the adult stage. A risk assessment procedure used previously was updated with a simulation model estimating the proportion of second-generation monarch butterflies affected. When considered over the entire range of the Corn Belt, which represents only 50% of the breeding population, the risk to monarch butterfly larvae associated with long-term exposure to Bt corn pollen is 0.6% additional mortality. Exposure also prolonged the developmental time of larvae by 1.8 d and reduced the weights of both pupae and adults by 5.5%. The sex ratio and wing length of adults were unaffected. The ecological significance of these sublethal effects is discussed relative to generation mortality and adult performance.

Setting the record straight: a rebuttal to an erroneous analysis on transgenic insecticidal crops and natural enemies

As scientists involved in risk assessment of transgenicinsecticidal plants, we are greatly concerned about thepublication by Lo¨vei et al. (2009) implying thatinsect-protected crops based on the Cry proteins ofBacillus thuringiensis may have substantial negativeimpacts on non-target organisms. We believe thatLo¨vei et al. (2009) use inappropriate and unsoundmethods for risk assessment that have led them toreach conclusions that are in conflict with those ofseveral recent comprehensive reviews and meta-analyses (e.g., O’Callaghan et al. 2005; Romeiset al. 2006; Marvier et al. 2007; Wolfenbarger et al.

Use of Transgenic Bacillus thuringiensis Berliner Corn Hybrids to Determine the Direct Economic Impact of the European Corn Borer (Lepidoptera: Crambidae) on Field Corn in Eastern Canada

Abstract Transgenic corn expressing Bacillus thuringiensis Berliner (Bt corn) (Maximizer and Yieldgard hybrids, Novartis Seeds), non-Bt isolines and high-performance (check) hybrids were evaluated for European corn borer, Ostrinia nubilalis (Hübner), damage and grain yield in commercial strip plots across Ontario in 1996 and 1997. Bt corn hybrids reduced stalk tunneling damage by 88–100%. In 1996, minimal damage was found in locations where only one generation of European corn borer occurred per year. Bt corn proved its greatest potential for reducing the number and length of cavities below the primary ear in locations where two generations of European corn borer were present. A yield response to using Bt hybrids only occurred when levels of tunneling damage exceeded 6 cm in length. European corn borer infestations resulted in a 6 and 2.4% reduction in yield for 1996 and 1997, respectively, when Bt hybrids were compared with their non-Bt isolines. A linear relationship was found between tunnel length per plant in centimeters (x) and yield protection (%) obtained from using Bt corn (y) (y = 1.02 + 0.005x, r2 = 0.7217). At a premium of

Quantitative analysis of 3-alkyl-2-methoxypyrazines in juice and wine using stable isotope labelled internal standard assay

34.58 Canadian (CDN) per hectare for Bt corn seed, an infestation of at least 6 cm of corn borer tunneling per plant was required to break even at a market price for corn of