Linking land use patterns and pest outbreaks in Bt maize.

Western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is a major pest of maize in the United States and is an invasive pest in Europe. Maize is the only agricultural crop on which western corn rootworm larvae can survive and this insect requires two consecutive years of maize cultivation to complete its life cycle. Transgenic maize producing insecticidal proteins derived from the bacterium Bacillus thuringiensis (Bt) is often used to manage rootworm populations. The first Bt trait, Cry3Bb1, was introduced in 2003, but larval resistance to this toxin appeared in northeastern Iowa in 2009. Rootworm management occurs on a field-by-field basis, but adult rootworm may disperse among fields. It is known that growing consecutive years of Cry3Bb1 maize within a field can lead to resistance, but the relationship of the surrounding landscape to the development of resistance is unknown. Using geospatial tools and publicly available land-use data, we examined circular areas (buffers) surrounding fields that had previously experienced high levels of rootworm injury to Cry3Bb1 maize and rootworm resistance to Cry3Bb1 maize (problem fields). We calculated the proportion of area inside each buffer planted to maize continuously for 1-9 yr, and compared these values to those for randomly selected control points throughout the state. We also calculated the proportion of the state planted to maize for at least three consecutive years for 2003 through 2018, and its relationship with the annual value of maize. We found that areas surrounding problem fields had significantly more continuous maize compared to controls, with the most continuous maize within 1.6 km of problem fields. We also found that the cultivation of continuous maize in Iowa increased significantly between 2003 and 2018, and this was correlated with average annual price of maize. We hypothesize a scenario in which continuous cultivation of Cry3Bb1 maize in local landscapes, driven in part by the increased value of maize, facilitated selection for Cry3Bb1 resistance. These results suggest that land use in areas surrounding problem fields affect the rate of resistance evolution and approaches for resistance management can be enhanced by taking a landscape-level perspective.

Applying a Selection Experiment to Test for Fitness Costs of Bt Resistance in Western Corn Rootworm (Coleoptera: Chrysomelidae) and the Effect of Density on Fitness Costs.

Western corn rootworm, Diabrotica virgifera virgifera LeConte, is a serious pest of corn and is often managed with transgenic corn producing insecticidal toxins from the bacterium Bacillus thuringiensis (Bt). This pest has developed field-evolved resistance to all commercially available Bt traits, beginning with Cry3Bb1 in 2009. Fitness costs may accompany Bt resistance, where individuals with alleles for Bt resistance have reduced fitness on non-Bt corn compared to Bt-susceptible individuals. In conjunction with non-Bt refuges, fitness costs can delay the evolution of Bt resistance. Importantly, ecological factors may affect the presence and magnitude of fitness costs. For western corn rootworm, available data suggest that fitness costs of Bt resistance may be present in some cases. Using two Cry3Bb1-resistant western corn rootworm strains (Hopkinton and Cresco), a fitness-cost experiment was performed by rearing rootworm in the absence of Bt for six generations to test for fitness costs of Cry3Bb1 resistance and the effect of larval rearing density on fitness costs. Fitness costs were detected for both strains; however, strains were still resistant to Cry3Bb1 corn at the end of the experiment. Cresco experienced a greater loss of resistance at low versus high density, but no effect of density was detected in Hopkinton. Our study shows that fitness costs can accompany Bt resistance in western corn rootworm and may be more pronounced under low larval density. Even though fitness costs were present, it appears that rootworm populations may remain resistant to Cry3Bb1 corn for years after resistance has evolved.

Western corn rootworm abundance, injury to corn, and resistance to Cry3Bb1 in the local landscape of previous problem fields.

Western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is a major pest of corn in the United States. Transgenic corn expressing insecticidal proteins derived from the bacterium Bacillus thuringiensis (Bt) is an important tool used to manage rootworm populations. However, field-evolved resistance to Bt threatens this technology. In areas where resistance is present, resistant individuals may travel from one field to a neighboring field, spreading resistance alleles. An important question that remains to be answered is the extent to which greater-than-expected root injury (i.e., >1 node of injury) to Cry3Bb1 corn from western corn rootworm is associated with rootworm abundance, root injury, and levels of resistance in neighboring fields. To address this question, fields with a history of greater-than-expected injury to Cry3Bb1 corn (focal fields) and surrounding fields (< 2.2 km from focal fields) were examined to quantify rootworm abundance, root injury, and resistance to Cry3Bb1 corn. Additionally, use of Bt corn and soil insecticide use for the previous six years were quantified for each field. Resistance to Cry3Bb1 was present in all fields assayed, even though focal fields had grown more Cry3 corn and less non-Bt corn than surrounding fields. This finding implies that some movement of resistance alleles had occurred between focal fields and surrounding fields. Overall, our data suggest that resistance to Cry3Bb1 in the landscape has been influenced by both local rootworm movement and field-level management tactics.

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.

Comparing Populations of Western Corn Rootworm (Coleoptera: Chrysomelidae) in Regions With and Without a History of Injury to Cry3 Corn.

Transgenic corn expressing insecticidal proteins derived from the bacterium Bacillus thuringiensis (Bt) is an important pest management tool. Western corn rootworm, Diabrotica virgifera virgifera LeConte, is a key pest of corn in the midwestern United States that has developed field-evolved resistance to all available Bt traits. The first Bt trait to be commercialized for management of rootworm was Cry3Bb1 in 2003, and field-evolved resistance appeared in 2009. In this study, we examined fields in counties where greater-than-expected injury to Cry3 (Cry3Bb1 or mCry3A) corn roots (>1 node) had previously been reported (problem counties) and counties where injury had not been reported (non-problem counties). Four to eight fields were sampled per county in 2015, 2016, and 2017 to quantify rootworm abundance, root injury, Cry3Bb1resistance, and rootworm management strategies. Rootworm abundance, root injury, and resistance to Cry3Bb1 did not differ between county types. Management tactics differed between county types, with problem counties growing more corn, using more soil insecticide, and growing more Cry34/35Ab1 corn. Additionally, a comparison of root injury to Bt and non-Bt corn within fields indicated that farmers derived an economic benefit from planting Bt corn to manage corn rootworm. Our results suggest that rootworm populations are similar between problem and non-problem counties in Iowa due to similar levels of selection pressure on Cry3 corn, but problem county fields have applied more management tactics due to previous rootworm issues in the area.

Combining in silico and in vitro models to inform cell seeding strategies in tissue engineering.

The seeding density of therapeutic cells in engineered tissue impacts both cell survival and vascularization. Excessively high seeded cell densities can result in increased death and thus waste of valuable cells, whereas lower seeded cell densities may not provide sufficient support for the tissue in vivo, reducing efficacy. Additionally, the production of growth factors by therapeutic cells in low oxygen environments offers a way of generating growth factor gradients, which are important for vascularization, but hypoxia can also induce unwanted levels of cell death. This is a complex problem that lends itself to a combination of computational modelling and experimentation. Here, we present a spatio-temporal mathematical model parametrized using in vitro data capable of simulating the interactions between a therapeutic cell population, oxygen concentrations and vascular endothelial growth factor (VEGF) concentrations in engineered tissues. Simulations of collagen nerve repair constructs suggest that specific seeded cell densities and non-uniform spatial distributions of seeded cells could enhance cell survival and the generation of VEGF gradients. These predictions can now be tested using targeted experiments.

Field-evolved resistance by western corn rootworm to Cry34/35Ab1 and other Bacillus thuringiensis traits in transgenic maize.

BACKGROUND: Transgenic crops producing insecticidal toxins from the bacterium Bacillus thuringiensis (Bt) are widely planted to manage agricultural insect pests. However, widespread adoption of Bt crops has led to the evolution of Bt resistance. The western corn rootworm, Diabrotica virgifera virgifera, is among the most serious pests of maize in the midwestern United States and is currently managed with Bt maize. To date, there is evidence of field-evolved resistance to all Bt toxins used to manage this pest. While western corn rootworm resistance to Cry3Bb1, and the closely related mCry3A and eCry3.1Ab traits, is widely distributed within the Midwest, fewer cases of Cry34/35Ab1 resistance have been observed, and planting of Cry34/35Ab1 maize is one of the methods used to manage Cry3-resistant rootworm. RESULTS: We found that fields with high levels of root injury to Cry34/35Ab1 maize by western corn rootworm were associated with Cry34/35Ab1-resistant western corn rootworm. Additionally, a population not associated with high levels of root injury was found to be resistant to Cry34/35Ab1. In all cases, populations that were resistant to Cry34/35Ab1 also were resistant to Cry3 traits. CONCLUSIONS: Western corn rootworm resistance to Cry34/35Ab1 has continued to persist in the agricultural landscape and has likely increased. The presence of rootworm populations with resistance to all available Bt traits threatens the utility of current and future transgenic technologies to manage this pest. Decreased reliance on Cry34/35Ab1 and better use of integrated pest management will be essential to preserve Bt susceptibility in western corn rootworm. © 2019 Society of Chemical Industry.

Atrazine Exposure Influences Immunity in the Blue Dasher Dragonfly, Pachydiplax longipennis (Odonata: Libellulidae).

Agricultural runoff containing herbicide is known to have adverse effects on freshwater organisms. Aquatic insects are particularly susceptible, and herbicide runoff has the potential to affect immunity in this group. Here we examined the effect of ecologically relevant levels of atrazine, an herbicide commonly used in the United States, on immune function in larvae of the blue dasher dragonfly (Odonata: Libelluludae, Pachydiplax longipennis Burmeister 1839) during a long-term exposure at ecologically relevant concentrations. Larvae were exposed to concentrations of 0, 1, 5, and 10 ppb atrazine for 3 or 6 wk. Hemocyte counts, hemolymph phenyloxidase (PO) activity, cuticular PO, and gut PO were measured at the end of each trial period as indicators of immune system strength. Atrazine concentration had a significant effect on hemocyte counts after controlling for larval size. There was a significant interaction between time and concentration for hemolymph PO, cuticular PO, and a marginal interaction for gut PO. The effect of atrazine on the measured immune parameters was often nonmonotonic, with larger effects observed at intermediate concentrations. Therefore, atrazine affects both hemocyte numbers and PO activity over time in P. longipennis, and the changed immune function demonstrated in this study is likely to modify susceptibility to pathogens, alter wound healing, and may decrease available energy for growth and metamorphosis.

Evidence of Resistance to Cry34/35Ab1 Corn by Western Corn Rootworm (Coleoptera: Chrysomelidae): Root Injury in the Field and Larval Survival in Plant-Based Bioassays.

Western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is a serious pest of corn in the United States, and recent management of western corn rootworm has included planting of Bt corn. Beginning in 2009, western corn rootworm populations with resistance to Cry3Bb1 corn and mCry3A corn were found in Iowa and elsewhere. To date, western corn rootworm populations have remained susceptible to corn producing Bt toxin Cry34/35Ab1. In this study, we used single-plant bioassays to test field populations of western corn rootworm for resistance to Cry34/35Ab1 corn, Cry3Bb1 corn, and mCry3A corn. Bioassays included nine rootworm populations collected from fields where severe injury to Bt corn had been observed and six control populations that had never been exposed to Bt corn. We found incomplete resistance to Cry34/35Ab1 corn among field populations collected from fields where severe injury to corn producing Cry34/35Ab1, either singly or as a pyramid, had been observed. Additionally, resistance to Cry3Bb1 corn and mCry3A corn was found among the majority of populations tested. These first cases of resistance to Cry34/35Ab1 corn, and the presence of resistance to multiple Bt toxins by western corn rootworm, highlight the potential vulnerability of Bt corn to the evolution of resistance by western corn rootworm. The use of more diversified management practices, in addition to insect resistance management, likely will be essential to sustain the viability of Bt corn for management of western corn rootworm.

Chronic sucrose intake reduces the antagonist effect of beta-funaltrexamine on morphine-induced antinociception in female but not in male rats.

Chronic ingestion of a sweet-tasting sucrose solution enhances the pain relieving actions of opioid agonists. These results, taken in conjunction with research demonstrating that sucrose stimulates the production and release of endogenous opioid peptides, have led to the hypothesis that the effects of palatable foods and fluids on pain sensitivity are mediated by the endogenous opioid system. To assess this hypothesis, two studies determined if chronic sucrose intake would block the antagonist effects of the micro-selective opioid antagonist beta-funaltrexamine (beta-FNA) on morphine-induced antinociception. Female and male Long-Evans rats were maintained on chow and water, or chow, water and a 32% sucrose solution. In Experiment 1, after four weeks on the diets, female rats received 0 or 10 mg/kg (subcutaneously (s.c.)) beta-FNA, while in Experiment 2, male and female rats received 0, 5 or 20 mg/kg beta-FNA. Six days later, rats were tested for morphine-induced antinociception using the hot-water tail-withdrawal test. Morphine, administered using a cumulative dose regime (1.0, 3.0, 5.6, 10.0 and 31.0 mg/kg s.c.), led to dose-dependent increases in tail-withdrawal latencies in male and female rats. Males were more sensitive to the pain relieving properties of morphine than females. Sucrose intake increased, while beta-FNA decreased the analgesic actions of morphine in males and females. beta-FNA was less effective in blocking the antinociceptive actions of morphine in sucrose-fed female rats than in females fed only chow. In contrast, diet had minimal effects on responses to beta-FNA in male rats.

Effect of pyruvate on the function of stallion spermatozoa stored for up to 48 hours.

Stallion spermatozoa maintain high fertilizing capacity if cooled to 5 degrees C and inseminated within 24 h. However, if spermatozoa are stored for 48 h, fertilizing capacity declines. Therefore, multiple shipments of semen are often required to inseminate mares that remain in estrus for days. Therefore, experiments were designed to determine if adding antioxidants to stallion spermatozoa stored at 5 degrees C for 48 h could maintain motility and fertilizing ability. In the first experiment stallion spermatozoa were incubated in a skim milk (SM) or a skim milk-egg yolk medium in combination with 10 mM pyruvate, 5 mM xanthurenic acid separately or in combination for up to 48 h at 5 degrees C. Spermatozoa incubated in SM for 48 h exhibited higher percentages of motile sperm (57%) than did sperm incubated in skim milk-egg yolk (34%); antioxidant treatment had little effect. In the second experiment, spermatozoa were incubated in SM containing 0, 1, 2, or 5 mM pyruvate. After 24 h of incubation at 5 degrees C, sperm incubated with 1, 2, or 5 mM pyruvate exhibited higher percentages of progressively motile spermatozoa (45%) than control exhibited (26%; P < 0.05). After 48 h, percentages of progressively motile spermatozoa were similar (27, 19, and 30 vs 14, respectively; P > 0.05). However, when incubated at 5 degrees C for 48 h and then incubated an additional 4 h at 25 degrees C, samples containing pyruvate exhibited higher percentages of motile (63 to 80%) and progressively motile (36 to 42%) sperm than did sperm in SM alone (28 and 5%, respectively; P < 0.05). The third experiment attempted to determine the optimal pyruvate concentration to maintain spermatozoal motility. Spermatozoa incubated with 0, 2, 3.5, or 5 mM pyruvate for 48 h at 5 degrees C and then an additional 4 h at 25 degrees C, exhibited similar percentages of progressively motile cells (31, 35, and 28%, respectively) that were higher than control (11%, P < 0.05). The last experiment evaluated the fertilizing potential of cooled spermatozoa. Embryos were recovered from 35, 20, and 30% of mares inseminated with spermatozoa that had been incubated at 5 degrees C, for 24 h in SM, or for 48 h in SM or SM + 2 mM pyruvate, respectively (P > 0.05). These studies indicate that 2 mM pyruvate in SM was beneficial in maintaining spermatozoal motility in 48 h-stored sperm and, although not significant, seemed to help maintain the fertility of 48 h-cooled spermatozoa.

A nicotine conjugate vaccine reduces nicotine distribution to brain and attenuates its behavioral and cardiovascular effects in rats.

Vaccination of animals to elicit drug-specific antibodies, or the passive transfer of such antibodies from other animals, can reduce the behavioral effects of drugs such as cocaine and heroin. To study the potential application of this approach to treating nicotine dependence, IgG was isolated from rabbits immunized with a nicotine-protein conjugate vaccine. Anesthetized rats received immune IgG containing nicotine-specific antibodies (Nic-IgG) or control-IgG i.v.. Thirty minutes later, rats received nicotine at 0.03 mg/kg i.v., equivalent on an mg/kg basis to the nicotine intake from two cigarettes by a smoker. Compared to control-IgG, Nic-IgG reduced the brain nicotine concentration in a dose-related manner (65% reduction at the highest IgG dose). Pretreatment with Nic-IgG also reduced the distribution to brain of five repeated doses of nicotine (equivalent to the nicotine intake from 10 cigarettes) administered over 80 min. To study blood pressure effects, rats received control-IgG or Nic-IgG 1 day prior to administering nicotine. Nicotine-induced systolic blood pressure increases were attenuated by Nic-IgG in a dose-related manner, and were almost completely blocked by the highest Nic-IgG dose. Pretreatment with Nic-IgG also completely prevented the nicotine-induced stimulation of locomotor activity observed in rats receiving control-IgG. Nic-IgG did not prevent locomotor activation from cocaine, demonstrating its specificity for nicotine. These data demonstrate that the administration of nicotine-specific antibodies can reduce or prevent some of the pharmacokinetic, cardiovascular, and behavioral consequences of nicotine in rats. Effects were observed at nicotine doses and nicotine serum concentrations equal to or exceeding those typically associated with nicotine exposure in cigarette smokers. A potential role for immunization in the treatment of nicotine dependence is suggested.

Drug form selection in albuterol-containing metered-dose inhaler formulations and its impact on chemical and physical stability.

New albuterol-containing metered-dose inhaler (MDI) formulations were under development to replace chlorofluorocarbon (CFC) propellants with more environmentally friendly hydrofluoroalkane (HFA) propellants. To achieve good chemical and physical stability of MDI formulations with HFA propellants, different drug forms were evaluated in model formulations (drug, oleic acid, and one of the following: P12/P11, P12/ethanol, P12, P134a/ethanol, P134a). The effects of drug form (base versus sulfate), propellant type (P12 versus P134a), and cosolvent type (P11 or ethanol versus none) on the chemical and physical stability were examined. The chemical stability of the formulations was determined by monitoring the percent drug remaining in the formulations using HPLC. The physical stability of the formulations was followed by visually assessing the suspension appearance, and by determining the mass median diameter (MMD) of the suspended particles using laser diffraction analysis. The drug form has a great impact on the chemical and physical stability of the formulations. The sulfate formulations were chemically stable up to 12 months when stored at 30 degrees C and 40 degrees C/85% relative humidity (RH). Poor chemical stability was observed for the base formulations, except for ethanol-free formulations (P12/P11, P12, and P134a) at 30 degrees C and a P134a formulation at 40 degrees C/85% RH. The chemical instability of albuterol base formulations at 30 degrees C correlates with its solubility. The presence of a cosolvent greatly improved the dispersion characteristics of both sulfate and base formulations. The sulfate formulations in the presence of a cosolvent (P12/P11, P12/ethanol, and P134a/ethanol) showed good physical stability when stored for up to 12 months at 30 degrees C and 40 degrees C/85% RH. The physical stability of the base formulations was not acceptable due to crystal growth/agglomeration in all formulations, except for the P12/P11 formulation. The physical instability of both sulfate and base formulations not only correlates with the drug solubility, but also with particle agglomeration. In conclusion, good chemical and physical stability of albuterol-containing suspension formulations can be achieved with the appropriate choice of drug form and formulation constituents.

Musculoskeletal etiology and therapy of craniomandibular pain and dysfunction.

A number of diverse symptoms, conventionally diagnosed and treated as individual problems, exist in combination in the craniomandibular pain and dysfunction patient. This chapter explores musculoskeletal dysfunction of the area as a common etiologic factor for head and neck pain. The electronically derived data, reported in 68 case histories, indicate that a common measurable etiology is responsible for the many ostensibly diverse manifestations of craniomandibular pain and dysfunction. Both quantitative and subjective data support the etiologic validity of the diagnosis. The basis for the effectiveness of the reported therapy rests on diagnosis of a skeletal malrelation of the mandible to the cranium that poses sustained accommodative demand on the neuromusculature.