Determining Factors and Economic Injury Levels for Sphenophorus levis for Chemical and Biological Control in Irrigated and Non-irrigated Sugarcane Crops.

Globally, people use sugarcane (Saccharum officinarum) to produce sugar and ethanol. Rainfed or irrigated sugarcane agricultural systems are available. Among the pests affecting this crop, the weevil Sphenophorus levis, Vaurie 1978 (Coleoptera: Curculionidae), is increasingly becoming a significant threat in southern South America. Sphenophorus levis populations are controlled using chemical or biological measures. Control decisions hinge upon the economic injury level (EIL). The EIL delineates the pest density that results in financial losses for producers. This study aims to determine the EIL for S. levis, considering the factors favoring this insect pest and chemical and biological control methods in rainfed and irrigated systems. The intensity of S. levis attacks was monitored in commercial sugarcane plantations over four years in João Pinheiro, Minas Gerais, Brazil. Sampling occurred in a 50 × 50 × 30-cm-deep trench dug in the soil surrounding the sugarcane clump. The total number of stumps in the clump, including those attacked by S. levis, was tallied. The EILs for this pest were 5.93% and 4.85% of targeted stumps for chemical control in rainfed and irrigated crops, respectively. Biological control in sugarcane plots resulted in an EIL of 4.15% and 3.40% for stumps attacked in rainfed and irrigated crops, respectively. Pest attacks were more severe during rainy years and in older sugarcane crops. The EIL values determined in this study could inform integrated pest management programs for sugarcane crops.

Fall armyworm (Lepidoptera: Noctuidae): practical resistance of 2 Brazilian populations to Cry1A.105†+†Cry2Ab and Cry1F Bt maize.

In the Americas, transgenic crops producing insecticidal proteins from Bacillus thuringiensis Berliner (Bt, Bacillales: Bacillaceae) have been used widely to manage fall armyworm (FAW, Spodoptera frugiperda [J.E. Smith]). As resistance to Cry1 single-gene Bt maize (Zea mays L.) rapidly evolved in some FAW populations, pyramided Bt maize hybrids producing Cry1, Cry2, or Vip3Aa proteins were introduced in the 2010s. We examined field-evolved resistance to single- and dual-protein Bt maize hybrids in 2 locations in southeastern Brazil, where plant damage by FAW larvae far exceeded the economic threshold in 2017. We collected late-instar larvae in Cry1A.105 + Cry2Ab and Cry1F maize fields and established 2 FAW populations in the laboratory. The F1 offspring reared on the foliage of Bt and non-Bt maize plants (Cry1A.105 + Cry2Ab and Cry1F) showed neonate-to-adult survival rates as high as 70% for both populations. There was no significant difference in the life-table parameters of armyworms reared on non-Bt and Bt maize foliage, indicating complete resistance to Cry1A.105 + Cry2Ab maize. Larval survival rates of reciprocal crosses of a susceptible laboratory strain and the field-collected populations indicated nonrecessive resistance to Cry1F and a recessive resistance to Cry1A.105 + Cry2Ab maize. When relaxing the selection pressure, the armyworm fitness varied on Cry1A.105 + Cry2Ab and non-Bt maize; the resistance was somewhat stable across 12 generations, without strong fitness costs, although one of the lines died confounded by a depleted-quality, artificial rearing diet. To our knowledge, this is the first report documenting the practical resistance of FAW to a pyramided Bt crop. We discuss the implications for resistance management.

Draft genome of neotropical Bacillus thuringiensis UFT038 and its potential against lepidopteran soybean pests.

Bacillus thuringiensis (Bt) is known for its Cry and Vip3A pesticidal proteins with high selectivity to target pests. Here, we assessed the potential of a novel neotropical Bt strain (UFT038) against six lepidopteran pests, including two Cry-resistant populations of fall armyworm, Spodoptera frugiperda. We also sequenced and analyzed the genome of Bt UFT038 to identify genes involved in insecticidal activities or encoding other virulence factors. In toxicological bioassays, Bt UFT038 killed and inhibited the neonate growth in a concentration-dependent manner. Bt UFT038 and HD-1 were equally toxic against S. cosmioides, S. frugiperda (S_Bt and R_Cry1 + 2Ab populations), Helicoverpa zea, and H. armigera. However, larval growth inhibition results indicated that Bt UFT038 was more toxic than HD-1 to S. cosmioides, while HD-1 was more active against Chrysodeixis includens. The draft genome of Bt UFT038 showed the cry1Aa8, cry1Ac11, cry1Ia44, cry2Aa9, cry2Ab35, and vip3Af5 genes. Besides this, genes encoding the virulence factors (inhA, plcA, piplC, sph, and chi1-2) and toxins (alo, cytK, hlyIII, hblA-D, and nheA-C) were also identified. Collectively, our findings reveal the potential of the Bt UFT038 strain as a source of insecticidal genes against lepidopteran pests, including S. cosmioides and S. frugiperda.

Cervical cytology results among pregnant and non-pregnant women in Brazil.

OBJECTIVE: To compare the prevalence of abnormal cervical smear results among pregnant and non-pregnant women with and without representation of the transformation zone (TZ) in a mid-sized city in Brazil. STUDY DESIGN: This observational analytical and retrospective study analyzed cervical smear results from pregnant and non-pregnant women in a mid-sized city in Brazil. Private data were collected at private clinics over seven years and from public services over 16 years. All results were reported according to Bethesda System. Pregnant women were separated from non-pregnant women, and women were stratified into two age groups (25-29 years and 30-34 years) in general analyses. For TZ representation and cytologic-histologic correlation, there was no age stratification. RESULTS: Among public services cervical smears, the frequency of atypical squamous cells of undetermined significance (ASC-US) was higher for pregnant women in the entire group (odds ratio [OR] 1.91; confidence interval [CI] 1.07-3.39) and among women from 30 to 40 years old (OR 2.79; CI 1.38-5.66). The same occurred for low-grade squamous intraepithelial lesion (LSIL) in the total group (OR 3.50; CI 1.44-8.53) and among women 30 to 40 years old (OR 4.57; CI 1.45-14.42). The frequency of ASC-US was higher in pregnant women with TZ representation exams than those without (OR 4.62; CI 2.17-9.84). The same occurred for those with atypical squamous cells, which cannot exclude high-grade squamous intraepithelial lesion (OR 9.81; CI 1.63-59.15) and LSIL (OR 10.47; CI 3.39-32.36). Among pregnant women with abnormal cervical smear results, 85.74% were followed up through cytology, and an average of 26.12% of patients underwent biopsy, of which almost all were positive for dysplasia. CONCLUSION: Pregnant and non-pregnant women did not differ in cervical smears with actual precursor lesions. Nevertheless, considering the opportunistic nature of Brazilian cervical cancer screening, prenatal visits present an opportunity to perform cervical smears in pregnant women. TZ representation correlated with higher rates of abnormal cervical smears, reinforcing the clinical importance of endocervical/metaplastic cell representation, including among pregnant women.

The role of genetically engineered soybean and Amaranthus weeds on biological and reproductive parameters of Spodoptera cosmioides (Lepidoptera: Noctuidae).

BACKGROUND: In soybean fields containing insecticide- and herbicide-resistant genetically engineered varieties, some weed species have increasingly become difficult to manage and may favor the population growth of secondary pests like Spodoptera cosmioides (Walker) (Lepidoptera: Noctuidae). To test this hypothesis, we measured life-history traits, population growth parameters and adult nutrient content of S. cosmioides reared on foliage from four Amaranthus species, from Cry1Ac Bt and non-Bt soybean varieties, and on meridic artificial diet. RESULTS: Larvae reared on A. palmeri and A. spinosus had a shorter development time (5-7 days) than larvae raised on the soybean varieties and A. hybridus. Armyworm survival probability was zero on A. viridis and highest (80% and 71%) on soybeans and A. palmeri. The latter and the artificial diet produced the heaviest larvae and pupae, in contrast to the non-Bt soybean variety. Body nutrient content diverged mostly for adults reared on artificial diet compared with those raised on the soybean varieties. The intrinsic rate of population increase (overall fitness) was 27.88% higher for the armyworms on A. palmeri, Cry1Ac Bt soybean and artificial diet compared with those on non-Bt soybean, A. spinosus and A. hybridus. CONCLUSIONS: Cry1Ac soybean fields infested by some Amaranthus weeds, especially A. palmeri, are conducive to the population growth of S. cosmioides. Integrated pest management programs may be needed to properly manage S. cosmioides in soybean fields, with surveillance for population peaks and judicious control measures when needed. © 2022 Society of Chemical Industry.

Seed treatment for managing fall armyworm as a defoliator and cutworm on maize: plant protection, residuality, and the insect life history.

BACKGROUND: The highly polyphagous and invasive fall armyworm (FAW, Spodoptera frugiperda) can feed on different plant parts of host crops, damaging whorls and stalks in early maize growth stages. Systemic insecticide seed treatment (IST) could minimize this damage, although the residual efficacy may vary with the plant tissue damaged. Using damage rating scales and artificial infestation in controlled conditions, we determined the potential of IST against FAW attacking maize whorl leaves or the stalk base. RESULTS: Chlorantraniliprole, cyantraniliprole, or thiodicarb + imidacloprid IST similarly killed > 80% FAWs for 1 or 2 weeks after plant emergence depending on the plant tissue attacked. The residual efficacy (i.e. time after plant emergence sustaining > 80% larval mortality) lasted from the first to the eleventh day (VE-V3 maize growth stages), while for cutworm on the maize stalk base, it lasted 3-7 days after plant emergence (V1-V2 stages). In terms of damage, the ISTs lasted 15 days after emergence (V4 stage) for FAW on whorl leaves and 10 days (V3 stage) for FAW feeding on the stalk base. The larvae surviving on the seed-treated plants underwent sublethal effects in growth and development, reducing insect fitness. CONCLUSION: Diamide or carbamate + neonicotinoid seed treatments kill FAW larvae on maize whorls or stalks in favorable edaphoclimatic and insecticide-susceptibility conditions. The cumulative impacts of systemic IST on aboveground insect pests go beyond mortality. The ISTs studied can be valuable against FAW in maize, for instance, to help protect varieties that may not express sufficient insect resistance in maize early growth stages.

Comparison of cervical cancer screening results among public and private services in Brazil.

OBJECTIVE: To compare the frequency of cervical smear test results between women seen in private and public health services in a medium-sized city in Brazil. METHODS: This was an observational analytical study analyzing cervical cytologic results over 16 years. Public data were collected from the Brazilian National Health System, and private refers to those collected at private clinics. RESULTS: The overall frequency of unsatisfactory/rejected results was higher in public service examinations (odds ratio [OR] 0.05; 95% confidence interval [CI] 0.04-0.06). The same occurred for atypical squamous cells of undetermined significance (ASC-US) (OR 0.90; 95% CI 0.85-0.95) and atypical squamous cells cannot exclude high grade intraepithelial lesions (ASC-H) (OR 0.55; 95% CI 0.47-0.64) categories. For low-grade squamous intraepithelial lesions, the frequency was higher among women from private services (OR 1.39; 95% CI 1.24-1.55). Among women aged 25-64 years, the frequency of high-grade squamous intraepithelial lesions was higher among women seen in the public service (OR 0.81; 95% CI 0.66-0.99). CONCLUSION: Brazilian public services showed higher frequencies of unsatisfactory/rejected results and higher rates of ASC-US and ASC-H. Focusing on pre-analytical phases and establishing an internal quality control program can help improve these rates even though national protocols guide them.

Insecticide seed treatment against corn leafhopper: helping protect grain yield in critical plant growth stages.

BACKGROUND: The corn leafhopper, Dalbulus maidis (Hemiptera: Cicadellidae), spreads maize stunt pathogens and requires timely and effective crop protection. We determined the interaction between maize phenology and the vector feeding/infection period by stunt pathogens with the residual efficacy of neonicotinoid insecticidal seed treatments. Greenhouse- and field-grown maize plants, seed-treated with clothianidin or imidacloprid insecticides, were infested during seven growth stages with corn leafhoppers reared under controlled conditions on maize plants displaying infection symptoms by both spiroplasma (corn stunt spiroplasma, Spiroplasma kunkelii) and phytoplasma (maize bushy phytoplasma) pathogens. RESULTS: In the greenhouse and field settings, seed treatment reduced the stunt disease symptoms and corn yield loss during the VE-V4 maize growth stages and caused no phytotoxicity. The neonicotinoid seed treatment reduced 20-60% of the yield losses from the corn stunt disease until the V4 growth stage. Infestation by infective corn leafhoppers in the V12 maize growth stage caused a 25-30% yield loss irrespective of seed treatment, yet no stunt disease symptom was evident. Nonetheless, corn yield losses and visual stunt symptoms as rated by a nine-category ordinal scale were strongly correlated (r = 0.79, P < 0.01). CONCLUSION: These results reinforce that maize plants are more susceptible to leafhopper stunt disease during the VE-V4 growth stages (emergence to the fourth-leaf stage). Seed treatment helps reduce the damage in the early growth stages (VE-V2), although supplemental control measures depending on leafhopper population density may be needed from VE-V12 to protect yield losses from the maize stunt condition. © 2021 Society of Chemical Industry.

Whole genome comparisons reveal panmixia among fall armyworm (Spodoptera frugiperda) from diverse locations.

BACKGROUND: The fall armyworm (Spodoptera frugiperda (J.E. Smith)) is a highly polyphagous agricultural pest with long-distance migratory behavior threatening food security worldwide. This pest has a host range of > 80 plant species, but two host strains are recognized based on their association with corn (C-strain) or rice and smaller grasses (R-strain). The population genomics of the United States (USA) fall armyworm remains poorly characterized to date despite its agricultural threat. RESULTS: In this study, the population structure and genetic diversity in 55 S. frugiperda samples from Argentina, Brazil, Kenya, Puerto Rico and USA were surveyed to further our understanding of whole genome nuclear diversity. Comparisons at the genomic level suggest a panmictic S. frugiperda population, with only a minor reduction in gene flow between the two overwintering populations in the continental USA, also corresponding to distinct host strains at the mitochondrial level. Two maternal lines were detected from analysis of mitochondrial genomes. We found members from the Eastern Hemisphere interspersed within both continental USA overwintering subpopulations, suggesting multiple individuals were likely introduced to Africa. CONCLUSIONS: Our research is the largest diverse collection of United States S. frugiperda whole genome sequences characterized to date, covering eight continental states and a USA territory (Puerto Rico). The genomic resources presented provide foundational information to understand gene flow at the whole genome level among S. frugiperda populations. Based on the genomic similarities found between host strains and laboratory vs. field samples, our findings validate the experimental use of laboratory strains and the host strain differentiation based on mitochondria and sex-linked genetic markers extends to minor genome wide differences with some exceptions showing mixture between host strains is likely occurring in field populations.

Plant Resistance in Some Modern Soybean Varieties May Favor Population Growth and Modify the Stylet Penetration of Bemisia tabaci (Hemiptera: Aleyrodidae).

Complaints of severe damage by whiteflies in soybean fields containing genetically engineered (GE) varieties led us to investigate the role of transgenic soybean varieties expressing resistance to some insects (Cry1Ac Bt toxin) and to herbicide (glyphosate) on the population growth and feeding behavior of Bemisia tabaci (Gennadius) MEAM1 (Hemiptera: Aleyrodidae). In the laboratory, the whiteflies reared on the GE Bt soybeans had a net reproductive rate (R0) 100% higher and intrinsic rate of population increase (rm) 15% higher than those reared on non-GE soybeans. The increased demographic performance was associated with a higher lifetime fecundity. In electrical penetration graphs, the whiteflies reared on the GE soybeans had fewer probes and spent 50% less time before reaching the phloem phase from the beginning of the first successful probe, indicating a higher risk of transmission of whitefly-borne viruses. Data from Neotropical fields showed a higher population density of B. tabaci on two soybean varieties expressing glyphosate resistance and Cry1Ac Bt toxin. These results indicate that some GE soybean varieties expressing insect and herbicide resistances can be more susceptible to whiteflies than non-GE ones or those only expressing herbicide resistance. Most likely, these differences are related to varietal features that increase host-plant susceptibility to whiteflies. Appropriate pest management may be needed to deal with whiteflies in soybean fields, especially in warm regions, and breeders may want to consider the issue when developing new soybean varieties.

Demographic Performance of Helicoverpa zea Populations on Dual and Triple-Gene Bt Cotton.

Insecticidal toxins from Bacillus thuringiensis (Bt) are valuable tools for pest management worldwide, contributing to the management of human disease insect vectors and phytophagous insect pests of agriculture and forestry. Here, we report the effects of dual and triple Bt toxins expressed in transgenic cotton cultivars on the fitness and demographic performance of Helicoverpa zea (Boddie)-a noctuid pest, known as cotton bollworm and corn earworm. Life-history traits were determined for individuals of three field populations from a region where H. zea overwintering is likely. Triple-gene Bt cotton cultivars that express Cry and Vip3Aa toxins killed 100% of the larvae in all populations tested. In contrast, dual-gene Bt cotton that express Cry1Ac+Cry1F and Cry1Ac+Cry2Ab allowed population growth with the intrinsic rate of population growth (rm) 38% lower than on non-Bt cotton. The insects feeding on Bt cotton plants that express Cry1Ac+Cry2Ab, Cry1Ac+Cry1F, or Cry1Ab+Cry2Ae exhibited reduced larval weight, survival rate, and increased development time. Additionally, fitness parameters varied significantly among the insect populations, even on non-Bt cotton plants, likely because of their different genetic background and/or previous Bt toxin exposure. This is the first report of the comparative fitness of H. zea field populations on dual-gene Bt cotton after the recent reports of field resistance to certain Bt toxins. These results document the population growth rates of H. zea from an agricultural landscape with 100% Bt cotton cultivars. Our results will contribute to the development and validation of resistance management recommendations.

Contrasting susceptibility of lepidopteran pests to diamide and pyrethroid insecticides in a region of overwintering and migratory intersection.

BACKGROUND: Pesticide resistance is a growing issue worldwide, and susceptibility of pest populations should be monitored in migratory intersection regions for successful resistance management. We determined the susceptibility of eight noctuid species from the Florida Panhandle to bifenthrin (pyrethroid) and chlorantraniliprole (diamide). Larvae from field and laboratory populations were exposed to commercial insecticide formulations using the leaf-dip method in concentration-mortality bioassays. RESULTS: The field populations of Helicoverpa zea (Boddie), Spodoptera frugiperda (Smith), S. eridania (Stoll), S. exigua (Hubner) and Chloridea virescens (Fabricius) had reduced susceptibility to bifenthrin compared with the laboratory populations. Resistance ratios to bifenthrin were as high as 10 071-fold in S. exigua and 436-fold in S. frugiperda, while there was no reduced susceptibility in Agrotis ipsilon (Hufnagel). The susceptibility to chlorantraniliprole was similar between the field and laboratory populations studied, except for S. exigua that exhibited 630-fold resistance to the diamide. The probit regression equations indicated that the larval mortality of S. exigua and S. frugiperda populations was <80% with bifenthrin at the concentration equivalent to the label rate. Likewise, the estimated mortality of S. exigua larvae with chlorantraniliprole at the label rate concentration was <80%. CONCLUSIONS: The lepidopteran pest populations tested were variable in susceptibility to bifenthrin by contrast to more consistent susceptibility to chlorantraniliprole. These results help in the choice of effective insecticides for integrated pest management and resistance management in cropping systems colonized by migratory lepidopteran pests from the U.S. Gulf Coast region. © 2020 Society of Chemical Industry.

Rethinking biorational insecticides for pest management: unintended effects and consequences.

Biorational insecticides are composed of natural products, including animals, plants, microbes, and minerals, or are their derivates. The use of biorational products for the management of insect pests has grown intensively in recent years, which has increased their popularity and share on the insecticide global market. Much of these recent increases in the use of biorational insecticides has been derived from the generalized perception that conventional insecticides have undesirable ecological and human health impacts. However, the idea of simply replacing synthetic compounds with biorational insecticides without considering their potential unintended effects can mislead their use and reduce the market life of such pest management tools. A systematic literature survey encompassing over 15 000 scientific manuscripts published between 1945 and 2019 reinforces the bias of focusing on studying the targeted effects while overlooking the potential detrimental effects of biorational products on human health and the environment (e.g. death and negative sublethal effects on pollinators and beneficial arthropods such as parasitoids and predators). Thus, the risks associated with biorational compounds (e.g. control failures, the evolution of resistance, shift in dominance, and outbreaks of secondary or primary pests) need to be revisited and the outcomes of such inquiry could be decisive for their future use in pest management programs. The shortcomings of regulatory processes, knowledge gaps, and the outlook for the use of the biorational products in pest management are discussed. © 2020 Society of Chemical Industry.

Comparative genomic analysis and mosquito larvicidal activity of four Bacillus thuringiensis serovar israelensis strains.

Bacillus thuringiensis serovar israelensis (Bti) is used to control insect vectors of human and animal diseases. In the present study, the toxicity of four strains of Bti, named T0124, T0131, T0137, and T0139, toward Aedes aegypti and Culex quinquefasciatus larvae was analyzed. The T0131 strain showed the highest larvicidal activity against A. aegypti (LC50 = 0.015 µg/ml) and C. quinquefasciatus larvae (LC50 = 0.035 µg/ml) when compared to the other strains. Furthermore, the genomic sequences of the four strains were obtained and compared. These Bti strains had chromosomes sizes of approximately 5.4 Mb with GC contents of ~35% and 5472-5477 putative coding regions. Three small plasmids (5.4, 6.8, and 7.6 kb) and three large plasmids (127, 235, and 359 kb) were found in the extrachromosomal content of all four strains. The SNP-based phylogeny revealed close relationship among isolates from this study and other Bti isolates, and SNPs analysis of the plasmids 127 kb did not reveal any mutations in δ-endotoxins genes. This newly acquired sequence data for these Bti strains may be useful in the search for novel insecticidal toxins to improve existing ones or develop new strategies for the biological control of important insect vectors of human and animal diseases.

Like Parents, Like Offspring? Susceptibility to Bt Toxins, Development on Dual-Gene Bt Cotton, and Parental Effect of Cry1Ac on a Nontarget Lepidopteran Pest.

An important step to devise appropriate pest management strategies for armyworms (Lepidoptera: Noctuidae) in Bacillus thuringiensis Berliner (Bt) crops is to determine the lethal, sublethal, and parental effects of Bt toxins on target and nontarget pest species. Here we documented the susceptibility of black armyworm, Spodoptera cosmioides (Walker), to three Cry toxins and its life-history traits feeding on dual-toxin Bt cotton and an artificial diet containing sublethal concentrations of Cry1Ac. In concentration-response bioassays, black armyworm larvae showed low susceptibility to Cry toxins, with 853 ng/cm2 as the lowest value estimated for the median lethal concentration (LC50). The decreasing rank of toxicity was Cry1F, Cry2Aa, and Cry1Ac. Foliage of dual-toxin Bt cotton varieties (Cry1Ac + Cry1F and Cry1Ab + Cry2Ae) caused higher larval mortality than Cry1Ac-expressing cotton. Black armyworms showed reduced larval weight when growing on the Cry1Ac-treated diet, yet they reached adulthood and produced offspring. Interestingly, these larvae were grown on the control diet and showed reduced weight gain associated with the toxin exposure of the previous generation, indicating a parental effect of the exposure to Cy1Ac. The reduced larval weight was recovered in later instars, and there was no significant change in the population fitness of the parental armyworms or their offspring. To our knowledge, this is the first study documenting the parental effects of Bt toxins in insects. These results advance our understanding of potential responses of nontarget species when exposed to Bt toxins and contribute to design pest management programs for armyworms and other nontarget lepidopteran species exposed to Bt crops.

Bt-induced hormesis in Bt-resistant insects: Theoretical possibility or factual concern?

The biphasic dose-response of a stressor where low amounts of a toxicant may stimulate some biological processes is a recent focus of attention in insecticide ecotoxicology. Nonetheless, the importance and management consequences of this phenomenon of pesticide-induced hormesis remain largely unrecognized. Curiously, the potential induction of hormesis by insecticidal proteins such as Bacillus thuringiensis toxins (i.e., Bt toxins), a major agriculture pest management tool of widespread use, has been wholly neglected. Thus, we aimed to circumvent this shortcoming while assessing the potential occurrence of hormesis induced by the Bt toxin Cry1Fa in its main target pest species - the fall armyworm Spodoptera frugiperda. Concentration-response bioassays were carried out in a Bt-susceptible and a Bt-resistant population providing the purified Cry1Fa toxin in artificial diet and recording the insect demographic parameters. As significant hormetic effect was detected in both populations with a significant increase in the net reproductive rate and the intrinsic rate of population growth, the potential occurrence of Bt-induced hormesis was subsequently tested providing the insects with leaves from transgenic Bt maize expressing the toxic protein. The performance of the Bt-resistant insects was not different in both maize genotypes, indicating that the leaf expression of the Bt protein did not promote hormesis in the resistant insects. Thus, despite the Bt-induced hormesis detected in the purified protein bioassays, the phenomenon was not detected with current levels of Bt expression in maize minimizing the risk of this additional efficacy constraint besides that of field occurrence of Bt resistance.

Complete Sequences of Two Plasmids Found in a Brazilian Bacillus thuringiensis Serovar israelensis Strain.

Plasmids play a crucial role in the evolution of bacterial genomes by mediating horizontal gene transfer. In this work, we sequenced two plasmids found in a Brazilian Bacillus thuringiensis serovar israelensis strain which showed 100% nucleotide identities with Bacillus thuringiensis serovar kurstaki plasmids.

Negative cross-resistance between structurally different Bacillus thuringiensis toxins may favor resistance management of soybean looper in transgenic Bt cultivars.

High adoption rates of single-gene Bacillus thuringiensis (Bt) Cry1Ac soybean impose selection pressure for resistance in the soybean looper, Chrysodeixis includens, a major defoliator in soybean and cotton crops. To anticipate and characterize resistance profiles that can evolve, soybean looper larvae collected from field crops in Brazil in 2013 were selected for resistance to Cry1Ac. Using two methods of selection viz., chronic exposure to Cry1Ac cotton leaves and the seven-day larval exposure to purified Cry1Ac on the artificial diet, 31 and 127-fold resistance was obtained in 11 and 6 generations of selection, respectively. The resistance trait had realized heritability of 0.66 and 0.72, respectively, indicating that most of the phenotypic variation in Cry1Ac susceptibility of the soybean looper larvae was due to additive genetic variation. The Cry1Ac-selected populations showed positive cross-resistance to Cry1Ab (6.7-8.7 fold), likely because these Bt toxins have a very similar molecular structure. Importantly, the Cry1Ac-selected populations became more susceptible to Cry2Aa and Cry1Fa, showing negative cross-resistance (up to 6-fold, P < 0.05). These results indicate that Cry1Ac, Cry1Fa, and Cry2A are compatible in a multi-toxin approach to minimize the risk of rapid adaptation of the soybean looper to Bt toxins.

Glyphosate is lethal and Cry toxins alter the development of the stingless bee Melipona quadrifasciata.

Brazil is the second largest producer of genetically modified plants in the world. This agricultural practice exposes native pollinators to contact and ingestion of Bacillus thuringiensis proteins (e.g. Cry toxins) from transgenic plants. Furthermore, native bees are also exposed to various herbicides applied to crops, including glyphosate. Little is known about the possible effects of glyphosate and Cry proteins on stingless bees, especially regarding exposure at an immature stage. Here, we show for the first time that glyphosate is lethal, and that Cry toxins (Cry1F, Cry2Aa) alter the development of the stingless bee Melipona quadrifasciata upon contamination of larval food. Glyphosate was very toxic to the bee larvae, killing all of them within only a few days of exposure. Bees treated with Cry2Aa proteins had a higher survival rate and were delayed in their development, compared to the negative controls. Those treated with the Cry1F protein also suffered delays in their development, compared to the negative controls. In conclusion, the proteins Cry1F, Cry2Aa, and the herbicide glyphosate were highly toxic to the stingless bee M. quadrifasciata, causing lethal or sublethal effects which can severely impair colony growth and viability, and reduce pollination ability.

Essential oil of Siparuna guianensis as an alternative tool for improved lepidopteran control and resistance management practices.

Although the cultivation of transgenic plants expressing toxins of Bacillus thuringiensis (Bt) represents a successful pest management strategy, the rapid evolution of resistance to Bt plants in several lepidopteran pests has threatened the sustainability of this practice. By exhibiting a favorable safety profile and allowing integration with pest management initiatives, plant essential oils have become relevant pest control alternatives. Here, we assessed the potential of essential oils extracted from a Neotropical plant, Siparuna guianensis Aublet, for improving the control and resistance management of key lepidopteran pests (i.e., Spodoptera frugiperda and Anticarsia gemmatalis). The essential oil exhibited high toxicity against both lepidopteran pest species (including an S. frugiperda strain resistant to Cry1A.105 and Cry2Ab Bt toxins). This high insecticidal activity was associated with necrotic and apoptotic effects revealed by in vitro assays with lepidopteran (but not human) cell lines. Furthermore, deficits in reproduction (e.g., egg-laying deterrence and decreased egg viability), larval development (e.g., feeding inhibition) and locomotion (e.g., individual and grouped larvae walking activities) were recorded for lepidopterans sublethally exposed to the essential oil. Thus, by similarly and efficiently controlling lepidopteran strains susceptible and resistant to Bt toxins, the S. guianensis essential oil represents a promising management tool against key lepidopteran pests.