A mixture toxicity approach for environmental risk assessment of multiple insect resistance genes.

Multiple substance considerations applied to chemical mixtures in ecological risk assessments can be logically extended to nontarget organism (NTO) risk assessment for pyramided trait crops expressing multiple insect resistance genes. A case instance is developed that considers a two-protein pyramid of Cry1F and Cry1Ac synthetic proteins expressed in cotton (Gossypium hirsutum L.). A mixture toxicity approach was used to arrive at the aggregated multisubstance potentially affected fraction (msPAF) of NTOs that may be at risk from exposure to Cry1F + Cry1Ac cotton in representative-use environments. Development of the msPAF for putative susceptible NTOs considered laboratory toxicity data for Lepidoptera expressed in terms of additive mixture toxicity as well as data on in planta expression of the Cry1F and Cry1Ac proteins and their translation into environmental loads and exposure concentrations. The msPAF based on tier 1 estimated environmental concentrations (EECs) and toxicity to Lepidoptera species-used as surrogate data for adverse effects to a putative susceptible species-provided a highly conservative estimate of effects on beneficial species and therefore is a ready means to conduct screening-level NTO risk assessments for pyramided crops.

Evaluating resistance to Bt toxin Cry1Ab by F2 screen in European populations of Ostrinia nubilalis (Lepidoptera: Crambidae).

The large-scale cultivation of transgenic crops producing Bacillus thuringiensis (Bt) toxins have already lead to the evolution of Bt resistance in some pest populations targeted by these crops. We used the F2 screening method for further estimating the frequency of resistance alleles of the European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae), to Bt maize, Zea mays L., producing the Cry1Ab toxin. In France, Germany, and Italy, 784, 455, and 80 lines of European corn borer were screened for resistance to Mon810 maize, respectively. In Slovakia, 26 lines were screened for resistance to the Cry1Ab toxin. The cost of F2 screen performed in the four countries varied from U.S. dollars 300 to dollars 1300 per line screened. The major difference in cost was mostly due to a severe loss of univoltine lines during the screen in Germany and Slovakia. In none of the screened lines did we detect alleles conferring resistance to Mon810 maize or to the Cry1Ab toxin. The frequency of resistance alleles were < 1.0 x 10(-3), < 1.6 x 10(-3), < 9.2 x 10(-3), and < 2.6 x 10(-2) in France, Germany, Italy, and Slovakia, with 95% probability, respectively. The average detection probability over all lines was approximately 90%. Making the assumption that European corn borer populations in these countries belong to the same genetic entity, the frequency of alleles conferring resistance to the Cry1Ab produced by the Mon810 maize in western and central Europe was 1.0 x 10(-4), with a 95% confidence interval of 0-3.0 x 10(-4).

Mating disruption of citrus leafminer mediated by a noncompetitive mechanism at a remarkably low pheromone release rate.

The citrus leafminer, Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae), is a worldwide pest of citrus. A season-long investigation was conducted that evaluated mating disruption for this pest. Effective disruption of the male P. citrella orientation to pheromone traps (98%) and reduced flush infestation by larvae was achieved for 221 d with two deployments of a 3:1 blend of (Z,Z,E)-7,11,13-hexadecatrienal/(Z,Z)-7,11-hexadecadienal at a remarkably low rate of 1.5 g active ingredient (AI)/ha per deployment. To gain insight into the mechanism that mediates the disruption of P. citrella, male moth catch was quantified in replicated plots of citrus treated with varying densities of pheromone dispensers. The densities of septum dispensers compared were: 0 (0/ha, 0.0 g AI/ha), 0.2 (one every fifth tree or 35/ha, 0.05 g AI/ha), 1 (215/ha, 0.29 g AI/ha), and 5 per tree (1,100/ha, 1.5 g AI/ha). Profile analysis by previously published mathematical methods matched predictions of noncompetitive mating disruption. Behavioral observations of male P. citrella in the field revealed that males did not approach mating disruption dispensers in any of the dispenser density treatments. The current report presents the first set of profile analyses combined with direct behavioral observations consistent with previously published theoretical predictions for a noncompetitive mechanism of mating disruption. The results suggest that disruption of P. citrella should be effective even at high population densities given the density-independent nature of disruption for this species and the remarkably low rate of pheromone per hectare required for efficacy.

Effects of gossypol on fitness costs associated with resistance to Bt cotton in pink bollworm.

Fitness costs associated with insect resistance to Bacillus thuringiensis (Bt) crops may help to delay or prevent the spread of resistance alleles, especially when refuges of non-Bt host plants are present. The potential for such delays increases as the magnitude and dominance of fitness costs increase. Here, we examined the idea that plant secondary chemicals affect expression of fitness costs associated with resistance to Bt cotton in Pectinophora gossypiella (Saunders). Specifically, we tested the hypotheses that gossypol affects the magnitude or dominance of fitness costs, by measuring performance of three independent sets of pink bollworm populations fed artificial diet with and without gossypol. Each set had an unselected susceptible population, a resistant population derived by selection from the susceptible population, and the F1 progeny of the susceptible and resistant populations. No individuals completed development on diets with gossypol in one set, suggesting that these individuals partially lost the ability to detoxify this chemical. In the other two sets, costs affecting survival did not support the hypotheses, but costs affecting pupal weight did. Adding gossypol to diet increased the magnitude and dominance of costs affecting pupal weight. In one of the two sets with survivors on diet with gossypol, costs affecting development time were less recessive when gossypol was present in diet. These results indicate that gossypol increased the magnitude and dominance of some fitness costs. Better understanding of the effects of natural plant defenses on fitness costs could improve our ability to design refuges for managing insect resistance to Bt crops.

Sequential sampling and biorational chemistries for management of lepidopteran pests of vegetable amaranth in the Caribbean.

Although vegetable amaranth, Amaranthus viridis L. and A. dubius Mart. ex Thell., production and economic importance is increasing in diversified peri-urban farms in Jamaica, lepidopteran herbivory is common even during weekly pyrethroid applications. We developed and validated a sampling plan, and investigated insecticides with new modes of action, for a complex of five species (Pyralidae: Spoladea recurvalis (F.), Herpetogramma bipunctalis (F.), Noctuidae: Spodoptera exigua (Hubner), S. frugiperda (J. E. Smith), and S. eridania Stoll). Significant within-plant variation occurred with H. bipunctalis, and a six-leaf sample unit including leaves from the inner and outer whorl was selected to sample all species. Larval counts best fit a negative binomial distribution. We developed a sequential sampling plan using a threshold of one larva per sample unit and the fitted distribution with a k(c) of 0.645. When compared with a fixed plan of 25 plants, sequential sampling recommended the same management decision on 87.5%, additional samples on 9.4%, and gave inaccurate recommendations on 3.1% of 32 farms, while reducing sample size by 46%. Insecticide frequency was reduced 33-60% when management decisions were based on sampled data compared with grower-standards, with no effect on crop damage. Damage remained high or variable (10-46%) with pyrethroid applications. Lepidopteran control was dramatically improved with ecdysone agonists (tebufenozide) or microbial metabolites (spinosyns and emamectin benzoate). This work facilitates resistance management efforts concurrent with the introduction of newer modes of action for lepidopteran control in leafy vegetable production in the Caribbean.

Assessment of Helicoverpa Armigera resistance to pyrethroid insecticides in northern Cameroon.

In Central Africa savannas, the noctuid Helicoverpa armigera (Hübner) is a major pest of cotton and vegetable crops (including tomato and okra) which directly affects production by attacking fruit-bearing organs. In order to prevent and anticipate problems, the national cotton company (SODECOTON), with the help of the agricultural research (IRAD-PRASAC), implemented in 1999 a monitoring network for the early detection of resistance of target insects to the main recommended insecticides. Over the last three years, from 1999 up to 2001, assessment of H. armigera susceptibility to pyrethroid insecticides was carried out through both vial tests and topical bioassays. Revealed in southern and western Africa, pyrethroid resistance now seems to affect Central Africa also. Although control failure due to resistance has not yet been reported from the field, these results indicate the presence of resistant genes within H. armigera populations in Northern Cameron. Further laboratory studies confirmed the loss of susceptibility to cypermethrin in wild populations. Priority actions to be developed must focus on the rational and concerted management of pesticide use and the implementation of a regional monitoring network for the early detection of the loss of sensibility of target insects to the main recommended insecticides. At the same time, more fundamental research should be undertaken on the epidemiological profile of resistance in order to define practical ways of reducing the selection pressure.

Assessment of two natural marine toxins (Microcystis aeruginosa and Parasicyonis actinostoloides) for the control of some medical and agriculture insects with reference to the action on mice.

The present work describes the insecticidal potentiality from Microcystis aeruginosa (cyanobacteria) and Parasicyonis actinostoloides (Sea anemone) water extracts against 3rd larval instar of medical insects (Musca domestica and Culex pipiens) and 4th larval instar of agriculture insects (Spodoptera littoralis and Agrotis ipsilon. The LD50s of M. aeruginosa extract were 1.94, 2.33, 7.59 and 9.10 mg/ml. for M. domestica, C. pipiens, S. littoralis and A. ipsilon). respectively. While LD50s of P. actinostoloides extract were 19.28, 24.7, 27.3 and (29.4) mg/ml. for M. domestica, C. pipiens, S. littoralis, and A. ipsilon respectively. The aqueous extracts of M. aeruginosa and P. actinostoloides had no acute or chronic marked effect on mice as serum acetyl cholinesterase and gave more or less nearly the same level of AchE activity at the end of decapitation periods.