Monarch Butterfly Ecology, Behavior, and Vulnerabilities in North Central United States Agricultural Landscapes.

The North American monarch butterfly (Danaus plexippus) is a candidate species for listing under the Endangered Species Act. Multiple factors are associated with the decline in the eastern population, including the loss of breeding and foraging habitat and pesticide use. Establishing habitat in agricultural landscapes of the North Central region of the United States is critical to increasing reproduction during the summer. We integrated spatially explicit modeling with empirical movement ecology and pesticide toxicology studies to simulate population outcomes for different habitat establishment scenarios. Because of their mobility, we conclude that breeding monarchs in the North Central states should be resilient to pesticide use and habitat fragmentation. Consequently, we predict that adult monarch recruitment can be enhanced even if new habitat is established near pesticide-treated crop fields. Our research has improved the understanding of monarch population dynamics at the landscape scale by examining the interactions among monarch movement ecology, habitat fragmentation, and pesticide use.

Optimization of QuEChERS Method for Simultaneous Determination of Neonicotinoid Residues in Pollinator Forage.

Consistent with the large-scale use of pesticide seed treatments in U.S. field crop production, there has been an increased use of neonicotinoid-treated corn and soybean seed over the past decade. Neonicotinoids can move downwind to adjacent off-field pollinator habitats in dust from planting and/or move downslope to habitats in surface water. The extent of potential neonicotinoid exposure to pollinators from neonicotinoid movement into these adjacent pollinator habitats is unclear. Pollen and leaf tissue extractions were completed using a quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction procedure. Samples were subjected to a clean-up step using dispersive solid-phase extraction (dSPE) techniques prior to analysis. The compounds in the extracts were separated on a reversed-phase column with gradient elution and confirmed with tandem mass spectrometry. The extraction method showed acceptable recoveries of analytes ranging from 78.4 to 93.6% and 89.4 to 101% for leaf tissue and pollen, respectively. The method's detection limits ranged from 0.04 to 0.3 ng/g in milkweed leaf tissue and 0.04 to 1.0 ng/g in pollen. The method is currently being employed in ongoing studies surveying pollen from a diversity of forbs and milkweed leaves obtained from habitat patches established within fields with a history of using neonicotinoid-treated seeds.

Uptake of C14-atrazine by prairie grasses in a phytoremediation setting.

Agrochemicals significantly contribute to environmental pollution. In the USA, atrazine is a widely used pesticide and commonly found in rivers, water systems, and rural wells. Phytoremediation can be a cost-effective means of removing pesticides from soil. The objective of this project was to investigate the ability of prairie grasses to remove atrazine. 14C-labeled atrazine was added to sterilized sand and water/nutrient cultures, and the analysis was performed after 21 days. Switchgrass and big bluestem were promising species for phytoremediation, taking up about 40% of the applied [14C] in liquid hydroponic cultures, and between 20% and 33% in sand cultures. Yellow Indiangrass showed low resistance to atrazine toxicity and low uptake of [14C] atrazine in liquid hydroponic cultures. Atrazine degradation increased progressively from sand to roots and leaves. Most atrazine taken up by prairie grasses from sand culture was degraded to metabolites, which accounted for 60-80% of [14C] detected in leaves. Deisopropylatrazine (DIA) was the main metabolite detected in sand and roots, whereas in leaves further metabolism took place, forming increased amounts of didealkylatrazine (DDA) and an unidentified metabolite. In conclusion, prairie grasses achieved high atrazine removal and degradation, showing a high potential for phytoremediation.

Comparison of the Insecticidal Characteristics of Commercially Available Plant Essential Oils Against Aedes aegypti and Anopheles gambiae (Diptera: Culicidae).

Aedes aegypti and Anopheles gambiae are two mosquito species that represent significant threats to global public health as vectors of Dengue virus and malaria parasites, respectively. Although mosquito populations have been effectively controlled through the use of synthetic insecticides, the emergence of widespread insecticide-resistance in wild mosquito populations is a strong motivation to explore new insecticidal chemistries. For these studies, Ae. aegypti and An. gambiae were treated with commercially available plant essential oils via topical application. The relative toxicity of each essential oil was determined, as measured by the 24-h LD(50) and percentage knockdown at 1 h, as compared with a variety of synthetic pyrethroids. For Ae. aegypti, the most toxic essential oil (patchouli oil) was ∼1,700-times less toxic than the least toxic synthetic pyrethroid, bifenthrin. For An. gambiae, the most toxic essential oil (patchouli oil) was ∼685-times less toxic than the least toxic synthetic pyrethroid. A wide variety of toxicities were observed among the essential oils screened. Also, plant essential oils were analyzed via gas chromatography/mass spectrometry (GC/MS) to identify the major components in each of the samples screened in this study. While the toxicities of these plant essential oils were demonstrated to be lower than those of the synthetic pyrethroids tested, the large amount of GC/MS data and bioactivity data for each essential oil presented in this study will serve as a valuable resource for future studies exploring the insecticidal quality of plant essential oils.

In Vitro Evaluation of Essential Oils and Saturated Fatty Acids for Repellency against the Old-World Sand Fly, Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae).

The sand fly, Phlebotomus papatasi (Scopoli, 1786), is a major vector for Leishmania major in the Middle East, which has impacted human health and US military operations in the area, demonstrating the need to develop effective sand fly control and repellent options. Here, we report the results of spatial repellency and avoidance experiments in a static air olfactometer using the female P. papatasi testing essential oils of Lippia graveolens (Mexican oregano), Pimenta dioica (allspice), Amyris balsamifera (amyris), Nepeta cataria (catnip), Mentha piperita (peppermint), and Melaleuca alternifolia (tea tree); the 9-12 carbon saturated fatty acids (nonanoic acid, decanoic acid, undecanoic acid, and dodecanoic acid); and the synthetic repellents DEET and IR3535. The materials applied at 1% exhibited varying activity levels but were not significantly different in mean repellency and avoidance from DEET and IR3535, except in regards to nonanoic acid. Some materials, particularly nonanoic and undecanoic acids, produced sand fly mortality. The observed trends in mean repellency over exposure time included the following: (1) P. dioica oil, M. alternifolia oil, decanoic acid, undecanoic acid, DEET, and IR3535 exhibited increasing mean repellency over time; (2) oils of N. cataria, A. balsamifera, M. piperita, and dodecanoic acid exhibited relatively constant mean repellency over time; and (3) L. graveolens oil and nonanoic acid exhibited a general decrease in mean repellent activity over time. These studies identified the essential oils of N. cataria and A. balsamifera as effective spatial repellents at reduced concentrations compared to those of DEET. Additional research is required to elucidate the modes of action and potential synergism of repellents and essential oil components for enhanced repellency activity.

Sorption and photodegradation processes govern distribution and fate of sulfamethazine in freshwater-sediment microcosms.

The antibiotic sulfamethazine can be transported from manured fields to surface water bodies. We investigated the degradation and fate of sulfamethazine in pond water using (14)C-phenyl-sulfamethazine in small pond water microcosms containing intact sediment and pond water. We found a 2.7-day half-life in pond water and 4.2-day half-life when sulfamethazine was added to the water (5 mg L(-1) initial concentration) with swine manure diluted to simulate runoff. Sulfamethazine dissipated exponentially from the water column, with the majority of loss occurring via movement into the sediment phase. Extractable sulfamethazine in sediment accounted for 1.9-6.1% of the applied antibiotic within 14 days and then declined thereafter. Sulfamethazine was transformed mainly into nonextractable sediment-bound residue (40-60% of applied radioactivity) and smaller amounts of photoproducts. Biodegradation, as indicated by metabolite formation and (14)CO2 evolution, was less significant than photodegradation. Two photoproducts accounted for 15-30% of radioactivity in the water column at the end of the 63-day study; the photoproducts were the major degradates in the aqueous and sediment phases. Other unidentified metabolites individually accounted for <7% of radioactivity in the water or sediment. Less than 3% of applied radioactivity was mineralized to (14)CO2. Manure input significantly increased sorption and binding of sulfamethazine residues to the sediment. These results show concurrent processes of photodegradation and sorption to sediment control aqueous concentrations and establish that sediment is a sink for sulfamethazine and sulfamethazine-related residues. Accumulation of the photoproducts and sulfamethazine in sediment may have important implications for benthic organisms.

Pan trapping soybean aphids (Hemiptera: Aphididae) using attractants.

Since its introduction into the United States in the past 10 yr, soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), has been a damaging pest to soybean, Glycine max (L.) Merrill. During 2008 and 2009, fields in central and north central Iowa experienced pockets of high soybean aphid populations. Electroantennograms have shown that soybean aphid alatae are capable of detecting host plant volatiles and sex pheromones. Here, we evaluated baited pan traps as a potential soybean aphid attractant. Yellow pan traps were placed in soybean fields after planting along with lures that contained plant volatiles and sex pheromones in 2008 or sex pheromones only in 2009. Pan trap contents were collected weekly, and plant counts also were conducted. Aphids were identified, and soybean aphids were counted to determine whether one chemical lure was more attractive to spring migrants than other lures. In both years, soybean aphids collected in pan traps with lures were not significantly different from the other products tested.

Bioallethrin activates specific olfactory sensory neurons and elicits spatial repellency in Aedes aegypti.

BACKGROUND: Use of pyrethroid insecticides is a pivotal strategy for mosquito control globally. Commonly known for their insecticidal activity by acting on voltage-gated sodium channels, pyrethroids, such as bioallethrin and transfluthrin, are used in mosquito coils, emanators and other vaporizers to repel mosquitoes and other biting arthropods. However, whether specific olfactory receptor neurons are activated by pyrethroids to trigger spatial repellency remains unknown. RESULTS: We used behavioral and electrophysiological approaches to elucidate the mechanism of bioallethrin repellency in Aedes aegypti, a major vector of dengue, yellow fever, Zika and chikungunya viruses. We found that bioallethrin elicits spatial (i.e. non-contact) repellency and activates a specific type of olfactory receptor neuron in mosquito antennae. Furthermore, bioallethrin repellency is significantly reduced in a mosquito mutant of Orco, an obligate olfactory co-receptor that is essential for the function of odorant receptors (Ors). These results indicate that activation of specific Or(s) by bioallethrin contributes to bioallethrin repellency. In addition, bioallethrin repellency was reduced in a pyrethroid-resistant strain that carries two mutations in the sodium channel gene that are responsible for knockdown resistance (kdr) to pyrethroids, indicating a role of activation of sodium channels in bioallethrin repellency. CONCLUSION: Results from this study show that bioallethrin repellency is likely to be the result of co-activation of Or(s) and sodium channels. These findings not only contribute to our understanding of the modes of action of volatile pyrethroids in spatial repellency, but also provide a framework for developing new repellents based on the dual-target mechanism revealed. © 2021 Society of Chemical Industry.

Optimization of analytical methods to improve detection of erythromycin from water and sediment.

Analytical methods to improve the detection of erythromycin in water and sediment were developed to optimize for erythromycin's recovery of extractable and bound residues from the aquatic environment. The objective of this study was to determine optimal recovery of erythromycin from water and sediment to improve its detection in environmental samples through solid-phase extraction (SPE) and sediment-extraction methods. SPE methods examined included previously reported methods for macrolide and sulfonamide antibiotics with erythromycin recoveries ranging from 75.5 % to 94.7 %. Extraction of erythromycin was performed from sand employing various solvents and buffers to determine the best method for extraction from two sandy loam pond sediments. Various extraction times were also examined, and all extraction procedures were performed in duplicate. The greatest recovery of (14)C-erythromycin in the Iowa sediment was 84 % using 0.3 M ammonium acetate at pH 4.2: acetonitrile (15:85, v/v) solution. The Oklahoma sediment yielded the greatest recovery of (14)C-erythromycin at 86.7 % with 0.3 M ammonium acetate at pH 7: acetonitrile (30:70, v/v) with a 60-minute shake time. The present results demonstrate improved extraction methods for enhancing the accuracy of erythromycin detection from environmental samples.

Evaluating toxicity of Varroa mite (Varroa destructor)-active dsRNA to monarch butterfly (Danaus plexippus) larvae.

Varroa mites (Varroa destructor) are parasitic mites that, combined with other factors, are contributing to high levels of honey bee (Apis mellifera) colony losses. A Varroa-active dsRNA was recently developed to control Varroa mites within honey bee brood cells. This dsRNA has 372 base pairs that are homologous to a sequence region within the Varroa mite calmodulin gene (cam). The Varroa-active dsRNA also shares a 21-base pair match with monarch butterfly (Danaus plexippus) calmodulin mRNA, raising the possibility of non-target effects if there is environmental exposure. We chronically exposed the entire monarch larval stage to common (Asclepias syriaca) and tropical (Asclepias curassavica) milkweed leaves treated with concentrations of Varroa-active dsRNA that are one- and ten-fold higher than those used to treat honey bee hives. This corresponded to concentrations of 0.025-0.041 and 0.211-0.282 mg/g leaf, respectively. Potassium arsenate and a previously designed monarch-active dsRNA with a 100% base pair match to the monarch v-ATPase A mRNA (leaf concentration was 0.020-0.034 mg/g) were used as positive controls. The Varroa mite and monarch-active dsRNA's did not cause significant differences in larval mortality, larval or pupal development, pupal weights, or adult eclosion rates when compared to negative controls. Irrespective of control or dsRNA treatment, larvae that consumed approximately 7500 to 10,500-mg milkweed leaf within 10 to 12 days had the highest pupal weights. The lack of mortality and sublethal effects following dietary exposure to dsRNA with 21-base pair and 100% base pair match to mRNAs that correspond to regulatory genes suggest monarch mRNA may be refractory to silencing by dsRNA or monarch dsRNase may degrade dsRNA to a concentration that is insufficient to silence mRNA signaling.

Monarch Butterfly (Danaus plexippus) Life-Stage Risks from Foliar and Seed-Treatment Insecticides.

Conservation of North America's eastern monarch butterfly (Danaus plexippus) population would require establishment of milkweed (Asclepias spp.) and nectar plants in the agricultural landscapes of the north central United States. A variety of seed-treatment and foliar insecticides are used to manage early- and late-season pests in these landscapes. Thus, there is a need to assess risks of these insecticides to monarch butterfly life stages to inform habitat conservation practices. Chronic and acute dietary toxicity studies were undertaken with larvae and adults, and acute topical bioassays were conducted with eggs, pupae, and adults using 6 representative insecticides: beta-cyfluthrin (pyrethroid), chlorantraniliprole (anthranilic diamide), chlorpyrifos (organophosphate), imidacloprid, clothianidin, and thiamethoxam (neonicotinoids). Chronic dietary median lethal concentration values for monarch larvae ranged from 1.6 × 10-3 (chlorantraniliprole) to 5.3 (chlorpyrifos) µg/g milkweed leaf, with the neonicotinoids producing high rates of arrested pupal ecdysis. Chlorantraniliprole and beta-cyfluthrin were generally the most toxic insecticides to all life stages, and thiamethoxam and chlorpyrifos were generally the least toxic. The toxicity results were compared to insecticide exposure estimates derived from a spray drift model and/or milkweed residue data reported in the literature. Aerial applications of foliar insecticides are expected to cause high downwind mortality in larvae and eggs, with lower mortality predicted for adults and pupae. Neonicotinoid seed treatments are expected to cause little to no downslope mortality and/or sublethal effects in larvae and adults. Given the vagile behavior of nonmigratory monarchs, considering these results within a landscape-scale context suggests that adult recruitment will not be negatively impacted if new habitat is established in close proximity of maize and soybean fields in the agricultural landscapes of the north central United States. Environ Toxicol Chem 2021;40:1761-1777. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Structure and Properties of Polyamide Fabrics with Insect-Repellent Functionality by Electrospinning and Oxygen Plasma-Treated Surface Coating.

The need for light-weight and high-strength insect-repellant fabrics is of critical importance to the cessation of viral diseases. The goal of the study is to investigate the structure and properties of insect-repellent polyamide fabrics for use in protective garments to guard against mosquitos. Permethrin was applied to the polyamide fabrics through incorporation into the nylon 6 polymer solution during electrospinning and dip coating onto the control untreated and oxygen plasma-treated polyamide fabrics: electropun nylon 6 nanofiber nonwovens, commercially available nylon 6 warp knit tricot, and nylon 66 double weft, knit interlock fabrics. The incorporation of permethrin into the polymer solution before the formation of fibers demonstrated the most efficient way to apply permethrin to the fiber/fabric systems. The plasma treatment significantly increased the amount of permethrin on the surface of the fabrics. All permethrin-containing polyamide fabrics showed excellent fastness of the insecticide to light. The electrospun nylon 6 nonwovens demonstrated the best fastness to washing among the plasma-treated electrospun nylon 6, nylon 66 double weft knit, and nylon 6 warp-knit tricot. All permethrin-treated fabrics were repellent and caused higher percentage of mosquito escape compared to the control untreated fabrics.

Assessing Field-Scale Risks of Foliar Insecticide Applications to Monarch Butterfly (Danaus plexippus) Larvae.

Establishment and maintenance of milkweed plants (Asclepias spp.) in agricultural landscapes of the north central United States are needed to reverse the decline of North America's eastern monarch butterfly (Danaus plexippus) population. Because of a lack of toxicity data, it is unclear how insecticide use may reduce monarch productivity when milkweed habitat is placed near maize and soybean fields. To assess the potential effects of foliar insecticides, acute cuticular and dietary toxicity of 5 representative active ingredients were determined: beta-cyfluthrin (pyrethroid), chlorantraniliprole (anthranilic diamide), chlorpyrifos (organophosphate), and imidacloprid and thiamethoxam (neonicotinoids). Cuticular median lethal dose values for first instars ranged from 9.2 × 10-3 to 79 µg/g larvae for beta-cyfluthrin and chlorpyrifos, respectively. Dietary median lethal concentration values for second instars ranged from 8.3 × 10-3 to 8.4 µg/g milkweed leaf for chlorantraniliprole and chlorpyrifos, respectively. To estimate larval mortality rates downwind from treated fields, modeled insecticide exposures to larvae and milkweed leaves were compared to dose-response curves obtained from bioassays with first-, second-, third-, and fifth-instar larvae. For aerial applications to manage soybean aphids, mortality rates at 60 m downwind were highest for beta-cyfluthrin and chlorantraniliprole following cuticular and dietary exposure, respectively, and lowest for thiamethoxam. To estimate landscape-scale risks, field-scale mortality rates must be considered in the context of spatial and temporal patterns of insecticide use. Environ Toxicol Chem 2020;39:923-941. © 2020 SETAC.

Biodistribution of degradable polyanhydride particles in Aedes aegypti tissues.

Insecticide resistance poses a significant threat to the control of arthropods that transmit disease agents. Nanoparticle carriers offer exciting opportunities to expand the armamentarium of insecticides available for public health and other pests. Most chemical insecticides are delivered by contact or feeding, and from there must penetrate various biological membranes to reach target organs and kill the pest organism. Nanoparticles have been shown to improve bioactive compound navigation of such barriers in vertebrates, but have not been well-explored in arthropods. In this study, we explored the potential of polyanhydride micro- and nanoparticles (250 nm- 3 µm), labeled with rhodamine B to associate with and/or transit across insect biological barriers, including the cuticle, epithelium, midgut and ovaries, in female Ae. aeygpti mosquitoes. Mosquitoes were exposed using conditions to mimic surface contact with a residual spray or paint, topical exposure to mimic contact with aerosolized insecticide, or per os in a sugar meal. In surface contact experiments, microparticles were sometimes observed in association with the exterior of the insect cuticle. Nanoparticles were more uniformly distributed across exterior tissues and present at higher concentrations. Furthermore, by surface contact, topical exposure, or per os, particles were detected in internal organs. In every experiment, amphiphilic polyanhydride nanoparticles associated with internal tissues to a higher degree than hydrophobic nanoparticles. In vitro, nanoparticles associated with Aedes aegypti Aag2 cells within two hours of exposure, and particles were evident in the cytoplasm. Further studies demonstrated that particle uptake is dependent on caveolae-mediated endocytosis. The propensity of these nanoparticles to cross biological barriers including the cuticle, to localize in target tissue sites of interest, and to reach the cytoplasm of cells, provides great promise for targeted delivery of insecticidal candidates that cannot otherwise reach these cellular and subcellular locations.

Evaluation of pyrethroids and organophosphates in insecticide mixtures for management of western corn rootworm larvae.

BACKGROUND: The western corn rootworm is an economically important pest of corn. Management tactics include pyrethroid and organophosphate insecticides, which may be applied as a mixture to protect corn roots. The goal of our study was to characterize the effects of pyrethroids and organophosphates alone and in combination on larval corn rootworm mortality and injury to corn roots. We evaluated two insecticide combinations: tebupirimphos with ß-cyfluthrin and chlorethoxyfos with bifenthrin. Using a soil-based, laboratory bioassay, we exposed larvae to five concentrations of the pyrethroid alone, the organophosphate alone, the combined formulation, and a water control. We calculated LC50 values and co-toxicity factors to determine synergism or antagonism between organophosphates and pyrethroids. We also measured adult emergence and root injury in a field experiment that tested tebupirimphos alone, ß-cyfluthrin alone, the combined formulation, and an untreated control. RESULTS: Bioassay results indicated antagonism between the pyrethroid and organophosphate at most concentrations for both insecticide combinations. In the field experiment, tebupirimphos alone or in combination with ß-cyfluthrin significantly reduced adult emergence and root injury compared to the untreated controls, but ß-cyfluthrin alone did not differ from the untreated control for either metric. CONCLUSIONS: These results suggest that, at the concentrations tested, the pyrethroid component of pyrethroid-organophosphate mixtures may not contribute to a reduction of rootworm emergence or root injury. While these pyrethroids may confer a management benefit for other pests, such as seedcorn maggot, the concentrations of pyrethroids present in current formulations of these mixtures are likely too low for effective rootworm management. © 2020 Society of Chemical Industry.

Laboratory evaluation of mobility and sorption for the veterinary antibiotic, tylosin, in agricultural soils.

Veterinary medicines, including antibiotics, are utilized in large quantities in intensive livestock farming. It is evidenced that tylosin, one of the most frequently used antibiotics, is only partially metabolized in animals and not completely degraded in the manure storage stage before application to the farmland. In order to assess the mobility of tylosin in soil, a soil-column leaching study and a simple batch sorption experiment were conducted in the laboratory. Tylosin had strong sorption to various soils, with sorption distribution coefficients ranging from 42 to 65 ml/g. The range of concentrations in leachate was detected from non-detectable to 0.27 ng/mL after four simulated rainfall events in one month, and leachability of tylosin is dependent on soil properties and manure amendment. Percentage of clay, organic matter, cation exchange capacity, and manure amendment were positively correlated with sorption, and negatively correlated with mobility of tylosin in soil. The majority of tylosin was not recovered in the testing system, indicating that tylosin was most likely mineralized, or irreversibly bound to solid particles since no major degradation products were detected. Some trace level tylosin residues from manure-applied farmlands may be the major source to surface water systems through soil erosion and preferential flow processes.

Menthol acts as a positive allosteric modulator on nematode levamisole sensitive nicotinic acetylcholine receptors.

The ongoing and widespread emergence of resistance to the existing anti-nematodal pharmacopeia has made it imperative to develop new anthelminthic agents. Historically, plants have been important sources of therapeutic compounds and offer an alternative to synthetic drugs. Monoterpenoids are phytochemicals that have been shown to produce acute toxic effects in insects and nematodes. Previous studies have shown nicotinic acetylcholine receptors (nAChRs) to be possible targets for naturally occurring plant metabolites such as carvacrol and carveol. In this study we examined the effects of monoterpenoid compounds on a levamisole sensitive nAChR from Oesophagostomum dentatum and a nicotine sensitive nAChR from Ascaris suum. We expressed the receptors in Xenopus laevis oocytes and used two-electrode voltage-clamp to characterize the effect of various compounds on these cys-loop receptors. At 100 µM the majority of these compounds acted as antagonists. Interestingly, further experiments revealed that both 0.1 µM and 10 µM menthol potentiated acetylcholine and levamisole responses in the levamisole sensitive receptor but not the nicotine sensitive receptor. We also investigated the effects of 0.1 µM menthol on the contractility of A. suum somatic muscle strips. Menthol produced significant potentiation of peak contractions at each concentration of acetylcholine. The positive allosteric modulatory effects of menthol in both in vivo and in vitro experiments suggests menthol as a promising candidate for combination therapy with cholinergic anthelmintics.

Aquatic fate and effects of Bacillus thuringiensis Cry3Bb1 protein: toward risk assessment.

Genetically engineered crops expressing Bacillus thuringiensis (Bt) insecticidal crystalline (Cry) proteins became commercially available in the United States in 1996. In 2006, 19 million ha of Bt corn were planted worldwide, which represents a 10 million ha increase in 10 years. The sustainability of Bt crops is important, because their use has significantly reduced the amount of chemical insecticides necessary to control agricultural pests. Despite the high adoption rates of this novel insecticide, little is known about the aquatic fate of transgenic Bt proteins and their nontarget effects on aquatic invertebrates, although several potential routes exist for their transport to aquatic systems. Methods were developed to investigate the aquatic fate of transgenic Bt proteins and to determine their potential effects on nontarget aquatic invertebrates. Laboratory microcosms containing pond water only or pond water and sediment were used to examine the fate of the coleopteran-active Bt Cry3Bb1 protein in decomposing transgenic corn event MON863 (hereafter referred to as MON863 corn) leaf, stalk, and root. A half-life of less than 3 d was found for Bt Cry3Bb1 from decomposing MON863 corn residue. No Bt Cry3Bb1 was measured in the pond water or sediment extracts of microcosms containing MON863 corn. In an acute, static, partial-renewal toxicity test, Bt Cry3Bb1 protein from MON863 root extracts was fed to Chironomus dilutus larvae for 10 d. A significant decrease in C. dilutus survival at nominal concentrations of 30 ng/ml was found; however, no effect on growth among the surviving larvae was observed.

Persistence and degradation of maize-expressed vaccine protein, Escherichia coli heat-labile enterotoxin subunit B, in soil and water.

Transgenic plants represent an innovative platform for the cost-effective large-scale production of various pharmaceutical proteins. The eventual open-field production of plant-made pharmaceuticals (PMPs) requires risk assessment to determine the potential for harm to the surrounding ecosystem. In the present study, the environmental persistence of a transgenic maize-expressed antigen, Escherichia coli heat-labile enterotoxin subunit B (LTB), was studied under laboratory conditions. To semiquantitatively monitor the persistence of LTB in soil, extraction with a high-salt, high-pH extraction buffer was optimized using the closely homologous Vibrio cholerae enterotoxin subunit B (CTB) as a test substance. The time to dissipation of 50% (DT50) of the extractable fraction of maize-expressed LTB was 4 to 15 d in pond water and 35 to 90 d in soils. Both extraction efficacy and persistence were strongly affected by the matrix type and incubation conditions. In contrast with maize-expressed LTB, the DT50 for bacterially produced LTB and CTB was less than 4 d both in pond water and soil. Although maize-expressed LTB was more stable than bacterially produced analogue, its dissipation was governed by an initial lag, which could be attributed to release from the plant material, followed by rapid decline.

Plant Essential Oils Enhance Diverse Pyrethroids against Multiple Strains of Mosquitoes and Inhibit Detoxification Enzyme Processes.

Mosquito-borne diseases account for the deaths of approximately 700,000 people annually throughout the world, with many more succumbing to the debilitating side effects associated with these etiologic disease agents. This is exacerbated in many countries where the lack of mosquito control and resources to prevent and treat mosquito-borne disease coincide. As populations of mosquito species grow more resistant to currently utilized control chemistries, the need for new and effective chemical means for vector control is more important than ever. Previous work revealed that plant essential oils enhance the toxicity of permethrin against multiple mosquito species that are of particular importance to public health. In this study, we screened permethrin and deltamethrin in combination with plant essential oils against a pyrethroid-susceptible and a pyrethroid-resistant strain of both Aedes aegypti and Anopheles gambiae. A number of plant essential oils significantly enhanced the toxicity of pyrethroids equal to or better than piperonyl butoxide, a commonly used synthetic synergist, in all strains tested. Significant synergism of pyrethroids was also observed for specific combinations of plant essential oils and pyrethroids. Moreover, plant essential oils significantly inhibited both cytochrome P450 and glutathione S-transferase activities, suggesting that the inhibition of detoxification contributes to the enhancement or synergism of plant essential oils for pyrethroids. This study highlights the potential of using diverse plant oils as insecticide additives to augment the efficacy of insecticidal formulations.