John J. Obrycki

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.

Tracking the role of alternative prey in soybean aphid predation by Orius insidiosus: a molecular approach

The soybean aphid, Aphis glycines (Hemiptera: Aphididae), is a pest of soybeans in Asia, and in recent years has caused extensive damage to soybeans in North America. Within these agroecosystems, generalist predators form an important component of the assemblage of natural enemies, and can exert significant pressure on prey populations. These food webs are complex and molecular gut‐content analyses offer nondisruptive approaches for examining trophic linkages in the field. We describe the development of a molecular detection system to examine the feeding behaviour of Orius insidiosus (Hemiptera: Anthocoridae) upon soybean aphids, an alternative prey item, Neohydatothrips variabilis (Thysanoptera: Thripidae), and an intraguild prey species, Harmonia axyridis (Coleoptera: Coccinellidae). Specific primer pairs were designed to target prey and were used to examine key trophic connections within this soybean food web. In total, 32% of O. insidiosus were found to have preyed upon A. glycines, but disproportionately high consumption occurred early in the season, when aphid densities were low. The intensity of early season predation indicates that O. insidiosus are important biological control agents of A. glycines, although data suggest that N. variabilis constitute a significant proportion of the diet of these generalist predators. No Orius were found to contain DNA of H. axyridis, suggesting intraguild predation upon these important late‐season predators during 2005 was low. In their entirety, these results implicate O. insidiosus as a valuable natural enemy of A. glycines in this soybean agroecosystem.

Uptake of Bt endotoxins by nontarget herbivores and higher order arthropod predators: molecular evidence from a transgenic corn agroecosystem.

The planting of transgenic crops expressing Bacillus thuringiensis endotoxins is widespread throughout the world; the prolific increase in their application exposes nontarget organisms to toxins designed to control pests. To date, studies have focused upon the effects of Bt endotoxins on specific herbivores and detritivores, without consideration of their persistence within arthropod food webs. Here, we report the first quantitative field evaluation of levels of Bt endotoxin within nontarget herbivores and the uptake by higher order arthropods. Antibody‐based assays indicated significant quantities of detectable Cry1Ab endotoxin within nontarget herbivores which feed on transgenic corn (including the corn flea beetle, Chaetocnema pulicaria, Japanese beetle, Popillia japonica and southern corn rootworm, Diabrotica undecimpunctata howardi). Furthermore, arthropod predators (Coccinellidae, Araneae, and Nabidae) collected from these agroecosystems also contained significant quantities of Cry1Ab endotoxin indicating its movement into higher trophic levels. This uptake by predators is likely to have occurred by direct feeding on plant material (in predators which are facultatively phytophagous) or the consumption of arthropod prey which contained these proteins. These data indicate that long‐term exposure to insecticidal toxins occurs in the field. These levels of exposure should therefore be considered during future risk assessments of transgenic crops to nontarget herbivores and arthropod predators.

HOST SPECIFICITY OF INSECT PARASITOIDS AND PREDATORS : MANY FACTORS INFLUENCE THE HOST RANGES OF INSECT NATURAL ENEMIES

BiO[OgiCa, control has beeil used more often against inseet pests than against any other graur of organisms. The most commonly used agents in these eHorts are ather inseets or related arthropods. These so-called beneficial speeies are classificd a5 eicher prcdators or parasiroids. Predators typically consumc several prey during their lives and can be predacious when immature, as adult:;, ur during both phases of thei! lives. ßy contrast, parasitoids live freely as adults but when immature feed in or on the body of an er her arthropod-the host-eventually killing ir. lnseet predators and parasitoids are referred to collectivelv as natural enemies. . Predators are distributed broadly across approximately 20 inseet orders (Ncw 1991). Parasitoids, by contrast, are more restricted taxonomically, occurring <:lImost cxelusively in the orders Hymenoptera (allts, bees, and wasps) and Diptera (flies; Clausen 1940). Nevertheless~ parasitoids are ineredibly abundant eomponents of aImost every ecosystem, both in terms of speeies and absolute nllmbers of individuals. Although estimates vary widely, rhe parasitie Hymenoptera may number

Impact of Transgenic Bacillus thuringiensis Corn and Crop Phenology on Five Nontarget Arthropods

Abstract Large-scale field studies were conducted to determine if temporal plantings of Bacillus thuringiensis (Berliner) (Bt) corn (event 176 and Bt11) would affect the seasonal abundance of the following generalist predators: Coleomegilla maculata DeGeer and Cycloneda munda (Say) (Coleoptera: Coccinellidae), Orius insidiosus (Say) (Heteroptera: Anthocoridae), Chrysoperla carnea Stephens (Neuroptera: Chrysopidae), and one specialist parasitoid, Macrocentrus cingulum Brischke (Hymenoptera: Braconidae). Adult populations were monitored using Pherocon AM yellow sticky traps at three locations in Iowa (1996–1998). At each location, a split-plot design was used with Bt and non-Bt corn as main plots and three planting dates as the split plots. Few differences in abundance were observed between Bt and non-Bt corn for the generalist predators studied. However, M. cingulum, a specialist parasitoid of European corn borer, was significantly affected by the presence of Bt corn. Densities of adult M. cingulum were 29–60% lower in Bt corn compared with non-Bt corn. Regression analyses indicated M. cingulum adults were preferentially recruited to and subsequently increased over time in the non-Bt corn treatments at each location within each year. Significant differences were observed among planting dates for all five species. Abundance effects from Bt corn on these natural enemies were not unexpected given the foraging and searching behaviors of different species and their varying levels of dependence on the presence of European corn borer.

Intraspecific alkaloid variation in ladybird eggs and its effects on con- and hetero-specific intraguild predators

Egg predation and cannibalism are common phenomena in predatory ladybirds despite the presence of defensive alkaloids. Consumption of heterospecific eggs negatively affects survivorship and development; however, intraspecific variation in quantities of alkaloids and post-ingestion responses to con- and hetero-specific alkaloids, are not well understood. We examined variation in the quantity of alkaloids in eggs of Harmonia axyridis (Pallas), Coccinella septempunctata L., and Hippodamia convergens (Guérin) using gas chromatography-mass spectrometry, and show a link between heterospecific alkaloids and their toxicity and/or costs by feeding high and low alkaloid eggs to first instar H. axyridis and C. septempunctata. The repeatability of alkaloid measurements in eggs in an egg cluster was high; however, the amount of alkaloids varied significantly between egg clutches within and among females. This variation affected egg consumption by C. septempunctata when fed H. axyridis eggs. Harmonia axyridis accumulated their own alkaloid by cannibalism and synthesized it de novo, but C. septempunctata lost some portion of the consumed conspecific alkaloids. Both species lost most of the consumed heterospecific alkaloids, but C. septempunctata died within 3xa0days. Most H. axyridis survived to the second instar, but C. septempunctata alkaloids led to a significant reduction in weight gain compared to an aphid control. In addition, ingestion of high alkaloid C. septempunctata extended development of H. axyridis compared to the aphid control or conspecific eggs. Harmonia axyridis had greater abilities to process ingested con- and hetero-specific alkaloids compared with C. septempunctata, which may, in part, explain their interspecific interactions in nature.

Larval Feeding on Bt Hybrid and Non-Bt Corn Seedlings by Harmonia axyridis (Coleoptera: Coccinellidae) and Coleomegilla maculata (Coleoptera: Coccinellidae)

Abstract Zoophytophagy is an omnivorous activity that occurs when a primarily carnivorous species feeds on plant material. Plant feeding by beneficial predators may have negative consequences if the plant material has been chemically treated, contains toxins, or was transgenically altered. Although common in predaceous Hemiptera, zoophytophagy has been rarely studied in aphidophagous coccinellids. This study examined the likelihood of feeding on Bt and non-Bt corn seedlings by third- and fourth-instar coccinellid larvae, the regularity of feeding events by fourth instars, and the effect of leaf feeding on development time and adult size. Both third- and fourth-instar Harmonia axyridis Pallas and Coleomegilla maculata DeGeer consume leaf tissue, with fourth instars being significantly more likely to feed on corn seedlings. C. maculata larvae ingested leaf tissue more frequently than H. axyridis. Furthermore, when given access to corn seedlings daily, development time of fourth-instar C. maculata increased after Bt hybrid corn treatments compared with non-Bt corn treatments. Zoophytophagous feeding behavior is thought to sustain predators during times of low prey availability, and leaf tissue feeding by coccinellids has typically been attributed to their need for water. However, in this study, tissue feeding regularly occurred even though coccinellid larvae had constant access to water and a daily ad libitum supply of aphids. We suggest that, in addition to environmental conditions, the physiological state of the zoophytophagous species will influence the probability of plant feeding.

Web-site selection strategies of linyphiid spiders in alfalfa: implications for biological control

Site-specific foraging can enhance the ability of generalist predators to provide effective and sustainable levels of pest control in agroecosystems. This can result from increased growth rates, higher population densities, and improved capture frequencies of pests at high prey density microsites. We tested the hypothesis that linyphiid spiders would exhibit microhabitat-specific web-site selection strategies in alfalfa. This was predicted to result in high prey densities at web-sites compared to paired non-web-sites through direct, or indirect, selection of prey-rich habitats. A total of 22,242 potential prey items were collected from mini-sticky traps located at 896 microsites. Web-centered mini-sticky traps on the ground, representative of Erigone autumnalis Emerton (Araneae: Linyphiidae) webs, captured similar numbers of potential prey as paired non-web-centered traps nearby. However, aerial sticky traps at web-sites of Bathyphantes pallidus (Banks) (Araneae: Linyphiidae) contained significantly more Diptera and Empoasca fabae (Harris) (Homoptera: Cicadellidae) than paired non-web-centered sticky-traps. Prey activity-densities also varied between web-sites of E. autumnalis and B. pallidus. Diptera were dominant at aerial microsites of B. pallidus whilst Collembola were abundant on ground-based traps of E. autumnalis. These results suggest that in alfalfa, the pressure for selecting prey-rich web-sites by erigonine spiders is low, but B. pallidus exhibits a selective web-location strategy targeted towards high quality dipteran prey. These sites also captured large numbers of E. fabae, a major pest of alfalfa, thus implicating aerial-based linyphiines as valuable predators in biological control.

Web-Construction Behavior of Linyphiid Spiders (Araneae, Linyphiidae): Competition and Co-Existence Within a Generalist Predator Guild

The web-construction behavior of three species of linyphiid spider (Erigone autumnalis, Meioneta unimaculata and Bathyphantes pallida) was studied in the laboratory to examine competition and co-existence within predator guilds. Competitive interactions between spiders potentially reduce their role in biological control. We tested the hypothesis that at high densities, intraguild competition for web-sites would occur but spatial separation of microhabitat would reduce interguild competition, thus allowing co-existence. High mortality and reduced web-size were observed at high B. pallida densities but Linyphiinae co-existed with Erigoninae which constructed webs at different strata. Competitive exclusion by larger individuals occurred between species whose microhabitat niche overlapped. The biocontrol potential of spider or arthropod predator guilds could ultimately be enhanced by maximizing the diversity of species whose niche axes vary.

Sugar feeding by coccinellids under field conditions: the effects of sugar sprays in soybean

Sucrose was applied weekly throughout the growing season at three US locations (South Dakota [SD], Maryland [MD], and Kentucky [KY]), and coccinellids and aphids (Aphis glycines Matsumura [Hemiptera: Aphididae]) were sampled 24xa0h later. Total coccinellid densities were 50–77% greater in sugar-sprayed soybean than in untreated plots. Coccinella septempuncata L., Hippodamia convergens Guérin-Méneville, and Harmonia axyridis Pallas were more abundant where sugar was applied. Coleomegilla maculata (DeGeer) was found in equally low numbers in all treatments. Aphid densities were similar in both treatments, and only reached economically threatening levels in SD. Coccinellids digested sugar meals within 1xa0h of consumption (measured using the cold anthrone test). Despite this narrow window of detection, field-collected coccinellids frequently tested positive for fructose. Under natural conditions, sugar is commonly ingested by coccinellids and sugar sprays increase coccinellid densities and their consumption of sugar. Sugar sprays did not enhance biological control of aphids in this experiment.