Álvaro Aguiar Coelho Teixeira

Fitness costs associated with field-evolved resistance to chlorantraniliprole in Plutella xylostella (Lepidoptera: Plutellidae)

Plutella xylostella (L.) is the most important pest of Brassicaceae worldwide, with a recent estimate of US

Morphology of the alimentary canal of Spodoptera frugiperda (J E Smith) Larvae (Lepidoptera: Noctuidae) fed on neem-treated leaves

4-5 billion expenditure for the control of this insect. A case of very high resistance of this pest to chlorantraniliprole was recently associated with reduced efficacy in a Brazilian field of Brassica spp. Although diamide resistance has been characterized, the fitness of insects due to such resistance has yet to be examined. Therefore, in this study, biological parameters were assessed in both susceptible and resistant strains of P. xylostella subjected to sublethal chlorantraniliprole concentrations. The field strain showed high resistance to chlorantraniliprole (RR50=27,793-fold), although resistance rapidly decreased in the first generations, showing instability. The exposure of susceptible and resistant larvae to their respective LC1, LC10, and LC25 values led to an increased duration of the larval and pupae phases and reduced weight in both strains; however, no significant differences in pupal viability across the treatments were observed. The resistant insects presented significantly lower larval weight and fecundity and higher larval and pupal periods, hatchability, and male longevity when not exposed to chlorantraniliprole, suggesting a fitness cost associated with resistance. In addition, resistant females showed a significantly higher egg-laying period and longevity at LC25, whereas the males lived longer at LC1. Chlorantraniliprole negatively impacted the biological parameters of both strains tested, although these effects were more relevant to the resistant insects. Resistant P. xylostella showed negative and positive biological trade-offs when compared with the susceptible individuals in both the absence and presence of chlorantraniliprole. Despite the important role that these trade-offs may play in the evolution of resistance to chlorantraniliprole, practical applications still depend on such information as the dominance of fitness costs and resistance.

Spermatogenesis, changes in reproductive structures, and time constraint associated with insemination in Podisus nigrispinus.

Research involving plants with insecticide activity evolved significantly in the last decades. Among these plants, the neem tree, Azadirachta indica, is commonly used against several insects, mainly Lepidoptera. The neem efficiency depends on the target insect and on the concentration used. A barrier against potential toxic agents ingested together with the food is the alimentary canal. Thus, this research aimed to describe the histology of the alimentary canal of Spodoptera frugiperda (J E Smith) larvae fed on leaves treated with neem (Neemseto) at a concentration of 0.5% and 1.0% and non treated, at different intervals (48, 96, 144, 192 and 240 h), by quantifying the regenerative cells and analyzing the secretion of the mesenteron histochemically. Larvae were immobilized at low temperatures (-4 degrees C), the alimentary canal was removed, fixed in Boüin s aqueous, embedded in paraplast and historesin, sectioned and stained with hematoxilin-eosin and periodic acid- Schiff. The histology of the alimentary canal of S. frugiperda was similar to other lepidopterans. Neem effects on morphology were seen only in the mesenteron, depending on the time and concentration used, such as: epithelium, reduction on regenerative cells and on the secretory activity in this region, confirmed by the histochemistry in both neem concentrations. These alterations were observed after 96 h at 1.0%, and 144 h at 0.5%. These results indicate that neem (Neemseto), at the concentrations studied, may be effective to control S. frugiperda because it promotes meaningful morphological alterations in the mesenteron.

Histopathology and ultrastructure of midgut of Alabama argillacea (Hübner) (Lepidoptera: Noctuidae) fed Bt-cotton

Males of the predatory stinkbug, Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae), accomplish long and multiple matings. We hypothesize that this behavior is due to time requirement for spermatozoa production and their transference to the females. Thus, this work investigated the effect of mating status of males and mating duration on spermatozoa transference to the females and the location of spermatozoa in the male reproductive tract during mating. On females, morphological alterations on female spermatheca and associated structures during a mating were investigated. Analyses of male reproductive tract showed presence of spermatozoa in the lumen of vas deferens was independent of mating status (ca. virgin, 0, 12 and 24 h after having a full mating), indicating continuous spermatogenesis which is supported by the absence of a seminal vesicle for spermatozoa storage. Female spermatheca had no changes associated with the duration of mating. However, females exhibited spermathecal elastic duct swelling by 30 min of mating duration. The success of males in filling the female spermatheca with spermatozoa depends on duration of mating. Thus, the results indicate that multiple mating is a requirement for reproductive success in the species by transference of spermatozoa and accessory substances stored in the female spermathecal duct. Likewise, the long mating is a male requirement to transfer materials in appropriate amount to the female but it is not dependent on spermatozoa alone.

Ultrastructure of the Alabama argillacea (Hübner) (Lepidoptera: Noctuidae) midgut.

The interaction of Cry toxins from Bacillus thuringiensis in the midgut of some insect larvae determines their efficacies as insecticides, due to the expression and availability of sites of action of the toxin in the midgut. Researches point out cases of resistance to Cry toxin due to alterations in the binding sites in columnar cell membrane. We analyzed the effects of Cry1Ac toxin expressed by Bt-cotton plants on Alabama argillacea midgut morphophysiology clarifying in levels of morphological and ultrastructural. Larvae in the 4th instar of A. argillacea after 20 min from ingesting Bt-cotton leaves expressing 0.183 ng of Cry1Ac exhibited ultrastructural and morphological modifications in the columnar cells with significant changes in the mitochondrial polymorphism, cytoplasmic vacuolization, microvillus and basal labyrinth. Expressive morphological alterations were also observed in the goblet cells indicating that the columnar cells are not the only target of the Cry1Ac toxin. The regenerative cells did not modify their structures and exhibited decrease in regeneration capacity. In conclusion, the ingestion of 0.183 ± 0.077 ng of Cry1Ac was enough to promote alterations in the columnar and goblet cells, besides reducing significantly the number of regenerative cells, which may have contributed to larval death. Nevertheless, further studies are necessary to determine the true cause of death.

Ultrastructure of Tuta absoluta parasitized eggs and the reproductive potential of females after parasitism by Metarhizium anisopliae.

The insect midgut has ultimately been the focus of researches tempting to control insect pests because alterations in the insect gut may affect not only its development, but also physiological events such as nutrient absorption and transformation. The objective of the present work was to describe morphologically, histochemically, and ultrastructurally the larva midgut of Alabama argillacea (Hübner) (Lepidoptera: Noctuidae), a cotton key pest in Brazil. Light and electronic transmission microscopy was used to obtain images from midgut sections of late fourth-instar larvae of A. argillacea. In general, the morphology, histochemistry, and ultrastructure characteristics of A. argillacea midgut follow that described in the literature for other lepidopteran species. However, the results showed a mitochondrial polymorphism and branched microvilli, which suggest an ultrastrucutural and physiological modification possibly associated with a high absorption and secretion activity by the columnar cells of this species. This intense activity may favor a faster response related to the action of ingested microbial agents and/or toxins, and can explain the high susceptibility of A. argillacea to the agents of control such as the toxin of Bacillus thuringiensis.

Effect of the combination of metformin hydrochloride and melatonin on oxidative stress before and during pregnancy, and biochemical and histopathological analysis of the livers of rats after treatment for polycystic ovary syndrome

Among the microorganisms used in biological control, the muscadine fungus Metarhizium anisopliae (Metsch.) Sorok. is produced and formulated world wide aiming to control pests from several agricultural crops. This work evaluated effects of M. anisopliae isolate UFRPE-6 on the fecundity and mortality of Tuta absoluta (Lepidoptera: Gelechiidae) females and the mechanism of infection on eggs. The infection of the females by the fungus did not affect their oviposition and fecundity; however it affected the survival with total and confirmed mortality of 54.2% and 37.14%, respectively. The eggs were treated with suspension at concentration of 10(6)conidia/mL. The analysis under scanning electron microscopy showed that conidia germination and penetration processes in the eggs of T. absoluta started within the period of 6h after the inoculation. Several hyphal bodies were observed from 12h and an intense extrusion of the mycelium covering all the external surface of the eggs 72h after inoculation. Despite its moderate activity in adults, the isolate URPE-6 of M. anisopliae showed promising in the control of T. absoluta due to its pathogenicity and virulence to eggs from this pest.

Microscopic Analysis of Spodoptera frugiperda (Lepidoptera: Noctuidae) Embryonic Development Before and After Treatment with Azadirachtin, Lufenuron, and Deltamethrin

The aim of the present study was to analyze the effect of a combination of metformin hydrochloride and melatonin on oxidative stress together with a biochemical and histopathological analysis of the livers of Wistar rats induced with PCOS. The results indicated that a combination of the drugs was more effective in the reduction of plasmatic levels of liver enzyme alanine aminotransferase, nitric oxide and total glutathione, and decreased the inflammatory response and histopathological damage, producing results that were significantly similar to animals from the control group. A mixture of the drugs produced more effective results against liver toxicity caused by PCOS, encouraging the normalization of biochemical parameters. During pregnancy, there was reduced oxidative stress compared to monotherapeutic use of these drugs. Interestingly, the combination of the drugs caused a physiological reaction similar to responses identified in healthy rats without induction of the PCOS control group. However, the clinical and physiological effectiveness of the combination should be further explored, especially with respect to the possible side effects on offspring.

Histological description of the midgut and the pyloric valve of Tropidacris collaris (Stoll, 1813) (Orthopetera: Romaleidae)

ABSTRACT The botanical insecticides, growth regulators, and pyrethroids have an effect on the biology of Spodoptera frugiperda (Smith). However, no emphasis has been given to the effect of these insecticides on embryonic development of insects, in histological level. Thus, this research aimed to examine by light and scanning electron microscopy S. frugiperda eggs and to describe the embryonic development, before and after immersion treatment, using commercial concentrations and lower concentrations than commercial ones, of the compounds lufenuron (Match), azadirachtin (AzaMax), and deltamethrin (Decis-positive control). For light microscopy semithin sections of eggs were used, and for scanning electron microscopy, images of the surface of eggs, treated and untreated with insecticides. The morphological characteristics of S. frugiperda eggs, in general, were similar to those described in the literature for most of the insects in the order Lepidoptera. Spherical eggs slightly flattened at the poles, with chorion, yolk, vitelline membrane, and embryo formation. In both microscopic analysis, we observed that insecticides acted immediately and independent of concentration, resulting absence, or incomplete embryo, presented yolk granules widely dispersed, without vitellophage formation, chorion disintegration, disorganized blastoderm, presenting vacuoles, yolk region with amorphous cells, and formation of completely uncharacterized appendages. Thus, we conclude that the compounds lufenuron and azadirachtin interfere on S. frugiperda embryonic development.

Effect of sub-lethal doses of Bacillus thuringiensis subsp. Aizawai and deltamethrin with regard to fertility and organ toxicity in pregnant albino rats

The present research describes the histology of the midgut, gastric caeca, and pyloric valve of Tropidacris collaris (Stoll, 1813), (Orthopetera: Romaleidae). We used light microscopy, staining (Gomoris trichrome and periodic acid-Schiff (PAS)), and a routine histological analysis method (hematoxilin-eosin). The insects were obtained from, and also bred in, the Laboratory of Entomology, Department of Biology, of the Rural Federal University of Pernambuco (UFRPE). The collected material was fixed in alcoholic Botüin and embedded in paraplast. The results demonstrated that the midgut wall is composed of an inner epithelial layer and two outer layers of striate muscles: one internal (circular) and the other external (longitudinal), with connective tissue between the muscle fibers. The epithelium is single-layered, with two cell types: regenerative and elongated columnar. The gastric caeca presents muscle layers similar to those of the midgut. Simple columnar epithelium lines the gastric caeca, which presents villi and projects towards the lumen. The pyloric valve is of striate muscle tissue, covered by a single epithelial-cell layer.