Thiazolidine derivatives: In vitro toxicity assessment against promastigote and amastigote forms of Leishmania infantum and ultrastructural study.

Neglected diseases, such as Leishmaniasis, constitute a group of communicable diseases that occur mainly in tropical countries. Considered a public health problem with limited treatment. Therefore, there is a need for new therapies. In this sense, our proposal was to evaluate in vitro two series of thiazolidine compounds (7a-7e and 8a-8e) against Leishmania infantum. We performed in vitro evaluations through macrophage cytotoxicity assays (J774) and nitric oxide production, activity against promastigotes and amastigotes, as well as ultrastructural analyzes in promastigotes. In the evaluation of cytotoxicity, the thiazolidine compounds presented CC50 values between 8.52 and 126.83 µM. Regarding the evaluation against the promastigote forms, the IC50 values ranged between 0.42 and 142.43 µM. Compound 7a was the most promising, as it had the lowest IC50. The parasites treated with compound 7a showed several changes, such as cell body shrinkage, shortening and loss of the flagellum, intense mitochondrial edema and cytoplasmic vacuolization, leading the parasite to cell inviability. In assays against the amastigote forms, the compound showed a low IC50 (0.65 µM). These results indicate that compound 7a was efficient for both evolutionary forms of the parasite. In silico studies suggest that the compound has good oral bioavailability. These results show that compound 7a is a potential drug candidate for the treatment of Leishmaniasis.

Schistosomicidal effect of divaricatic acid from Canoparmelia texana (Lichen): In vitro evaluation and ultrastructural analysis against adult worms of Schistosoma mansoni.

In this study we evaluated the in vitro effect of divaricatic acid against coupled worms of Schistosoma mansoni. The schistosomicidal effect was evaluated through the bioassay of motility and mortality, cellular viability of the worms and ultrastructural analysis through Scanning Electron Microscopy. To evaluate the cytotoxicity of divaricatic acid, a cell viability assay was performed with human peripheral blood mononuclear cells. Divaricatic acid proved effect against S. mansoni after 3 hours of exposure. At the end of 24 h the concentrations of 100 - 200 µM presented lethality to the worms. Motility changes were observed at sublethal concentrations. The IC50 obtained by the cell viability assay for S. mansoni was 100.6 µM (96.24 - 105.2 µM). Extensive damage to the worm's tegument was observed such as peeling, erosion, bubbles, edema, damage and loss of tubercles and spines, fissures and tissue ruptures. No cytotoxicity was observed in human peripheral blood mononuclear cells. This report provides data showing the schistosomicidal effect of divaricatic acid on S. mansoni, causing death, motile changes and ultrastructural damage to worms. In addition, divaricatic acid was shown to be non-toxic to human peripheral blood mononuclear cells at concentrations effective on S. mansoni.

In vitro and in vivo effects of P-MAPA immunomodulator on schistosomiasis.

Schistosomiasis is an infectious disease caused by helminth parasites of the genus Schistosoma; it is transmitted in over 78 countries. The main strategy for schistosomiasis control is treatment of infected people with praziquantel (PZQ). As PZQ-resistant strains have emerged, new anti-schistosomal agents have become necessary. We evaluated the in vitro and in vivo effect of P-MAPA, an aggregated polymer of protein magnesium ammonium phospholinoleate-palmitoleate anhydride with immunomodulatory properties; it is produced by Aspergillus oryzae fermentation. In vitro, P-MAPA (5, 50, and 100 µg/mL) damaged the Schistosoma mansoni tegument, causing thorn losses and tuber destruction in male worms and peeling and erosion in females after 24-h incubation. In vivo, P-MAPA (5 and 100 mg/kg, alone and combined with PZQ - 50 mg/kg) reduced the number of eggs by up to 69.20% in the liver and 88.08% in the intestine. Furthermore, granulomas were reduced up to 83.13%, and there was an increase in the number of dead eggs and a reduction of serum aspartate aminotransferase levels. These data suggest that P-MAPA activity can help improve schistosomiasis treatment and patients' quality of life.

Evaluation of Piper marginatum (Piperales: Piperaceae) Oil and Geraniol on the Embryonic Development of Spodoptera frugiperda (Lepidoptera: Noctuidae) in Comparison to Formulated Products.

Essential oils and their isolated constituents are constantly being studied for the control of insect pests. In this context, the present research reports the chemical composition of Piper marginatum (Jacq.) oil aiming to: 1) establish lethal concentrations LC30 and LC50 for this oil and the compound geraniol, 2) histologically examine the embryonic development of Spodoptera frugiperda (J.E. Smith) through light microscopy and scanning electron microscopy (SEM), as well as 3) compare the efficacy of the P. marginatum oil with that of the botanical insecticide azadirachtin, the synthetic insecticide deltamethrin, and acetone as a negative control. Semithin sections of S. frugiperda eggs revealed that the oil, geraniol, azadirachtin, and deltamethrin affected embryonic development at both concentrations. However, geraniol and the oil were more efficient because they caused more significant damage, even at lower concentrations. SEM revealed that all products altered the morphology of the eggs, modifying the structure of the chorion and making the eggs nonviable. Thus, this work demonstrates that P. marginatum oil is effective in the control of S. frugiperda because it results in embryonic damage even at the lowest concentrations.

Ovicidal lectins from Moringa oleifera and Myracrodruon urundeuva cause alterations in chorionic surface and penetrate the embryos of Aedes aegypti eggs.

BACKGROUND: Lectins from Moringa oleifera seeds (WSMoL), Myracrodruon urundeuva bark (MuBL), and heartwood (MuHL) are larvicidal agents against Aedes aegypti; in addition, WSMoL is an ovicidal agent against this mosquito. In this work, we evaluated the ovicidal activity of MuBL and MuHL by determining the concentrations that reduce the hatching rates by 50% in 72 h (EC50 ). The effects of WSMoL, MuBL, and MuHL on the ultrastructure of the eggs' surface were assessed by scanning electron microscopy (SEM). In addition, the ability of these lectins to penetrate the eggs was investigated by using conjugates of the lectins with fluorescein isothiocyanate (FITC). RESULTS: MuBL and MuHL were ovicidal agents with EC50 of 0.26 and 0.80 mg/mL (260 and 800 ppm), respectively. SEM images of eggs treated with WSMoL for 24 h revealed discontinuity of the exochorionic network and the absence of the exochorionic cells and their tubercles. After 48 and 72 h of incubation, strong deformation and degeneration of egg surfaces were observed. In MuBL and MuHL-treated eggs, the presence of lumps on the surface of the eggs, disappearance of the exochorionic network and the decrease and deformation of tubercles were observed. Lastly, fluorescence microscopy revealed that the three lectins were able to enter the eggs and reach the digestive tract of the embryos. CONCLUSION: WSMoL, MuBL, and MuHL are ovicidal agents on A. aegypti that have differing efficiencies in terms of how they cause alterations in the chorionic surface and in terms of their ability to penetrate the eggs. © 2019 Society of Chemical Industry.

In vitro activity of usnic acid potassium salt against different developmental stages of Schistosoma mansoni: An ultrastructural study.

Currently, the control of schistosomiasis is based on a single drug, praziquantel, which is effective against all species of Schistosoma but only in the adult stage, presenting a schistosomicidal deficit at the other developmental stages of the parasites. Recently our research group has demonstrated that the potassium salt of usnic acid (PS-UA) presented schistosomicidal property against couples of adult worms of S. mansoni. Thus, the present study seeks to report for the first time the in vitro activity of PS-UA against different developmental stages of S. mansoni (schistosomules and young worms). As schistosomicide parameters, we evaluated motility, mortality, cell viability of the worms and tegument changes by scanning electron microscopy (SEM). After 3 h exposure, PS-UA was lethal to schistosomules at concentrations of 100 and 50 µM, whereas for concentrations 25 and 12.5 µM, 38 and 18% of mortality and 62 and 24% changes in motility, respectively, were reached. Yet for schistosomules, concentration of 25 µM caused 90 and 100% of death after 6 and 12 h, respectively. In the concentration of 12.5 µM at intervals of 12 and 24 h mortality was 68 and 100%, respectively. For young worms, after 3 h of exposure at concentrations of 200 and 100 µM caused 57 and 27% mortality, respectively. After 12 and 24 h, these concentrations caused mortality of 90 and 100% and 47 and 60% respectively. After 24 h, concentrations of 50 and 25 µM caused 80 and 30% change in motility, respectively. However, at the 12.5 µM concentration no change was observed. In addition, PS-UA reduced the cellular viability of young worms by 50.98% and 85.87% at concentrations of 100 and 200 µM, respectively. In both stages of worms and at different exposure intervals, PS-UA caused alterations such as: dorsoventral contraction, peeling, swelling, blisters, erosion, exposure of subtegumental tissue and disintegration of tegument. According to the results, changes in motility and mortality caused by PS-UA against schistosomules and young worms were concentration and time-dependents, also PS-UA even at low concentration, was able to cause profound ultrastructural changes in the integument of the worms. PS-UA is a promising candidate as prophylactic agent in the control of schistosomiasis mansoni.

Anatomy, histochemistry and oxalic acid content of the leaflets of Averrhoa bilimbi and Averrhoa carambola

Abstract Averrhoa bilimbi L. and A. carambola L., Oxalidaceae, are the only two species of the genus Averrhoa L. Their leaves are widely used in folk medicine as an adjuvant in the treatment of diabetes. Some species may contain, for example, calcium oxalate crystals, which may lead to risk in its use when there is predisposition of individuals with reduced renal activity. Therefore, there are still few studies on the content of oxalic acid present in them, highlighting the importance of this investigation. The objective of this work was to conduct a comparative anatomical and histochemical study between the species and determining its content of oxalic acid. Semipermanent histological slides were prepared, following common plant anatomy procedures, for analysis of the leaflets in optical microscopy, polarization and scanning electron microscopy coupled with energy-dispersive X-ray spectrometry. To determine the total, soluble and insoluble oxalate content was used titration with potassium permanganate. The anatomical characterization allowed identifying the characters useful in the differentiation of the species. The histochemistry revealed the location of the metabolites. Chemical microanalyses demonstrated that the crystals are of calcium oxalate. A. carambola presented the highest levels of total oxalate and soluble oxalate. The study assists in the identification and quality control of A. bilimbi and A. carambola and brings new data on its oxalic acid content, which are important, in view of the medicinal use of the species.

Usnic acid potassium salt from Cladonia substellata (Lichen): Synthesis, cytotoxicity and in vitro anthelmintic activity and ultrastructural analysis against adult worms of Schistosoma mansoni.

We report for the first time the in vitro effect of Potassium Salt, derived from Usnic Acid (PS-UA), isolated from the lichen Cladonia substellata Vanio, on couples of Schistosoma mansoni. As schistosomicide parameters, we evaluated mortality, motility, cell viability of the worms and tegument changes by scanning electron microscopy (SEM). Exposure to a concentration of 100 µM caused 75% mortality after 3 h. After 6 h, changes in motility in concentrations of 50 and 25 µM are evidenced. After 12 h and 24h, the concentrations of 50 and 100 µM caused 6.25% and 87.5% and 50% and 100% mortality, respectively. PS-UA reduced the cell viability of the worms by 27.36% and 52.82% at concentrations 50 and 100 µM, respectively. Through SEM we observed progressive dose-and time-dependent, alterations such as swelling, blisters, dorsoventral contraction, erosion until disintegration of the tubercles in the tegument of male and female. PS-UA did not alter the viability of human peripheral blood mononuclear cells and showed high selectivity indices (IC50 > 200 µM). Our results indicate that PS-UA represents a possible candidate for a new anthelmintic drug in the control of schistosomiasis.

Assessing the cellular and humoral immune response in Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) infected with Leishmania infantum (Nicolle, 1908).

The aim of this study was to evaluate aspects of the innate cellular and humoral immune response by evaluating hemocyte dynamics, phagocytosis, phenoloxidase (PO) activity and nitric oxide (NO) production in Rhipicephalus sanguineus sensu lato (s.l.) (Acari: Ixodidae) infected with Leishmania infantum and to assess the persistence of parasites at time 0 and 1, 2, 5, and 7 days post-infection (dpi). The total and differential count of the five types of hemocytes circulating in the hemolymph of R. sanguineus s.l. females showed the average total number of hemocytes in the group infected with L. infantum to be significantly higher (p < 0.05) on the 1st and 2nd dpi compared to the control group. The hemocyte differential count showed that the average number of plasmatocytes and granulocytes increased significantly on the 1st, 2nd, and 5th dpi with L. infantum compared to the control group (p < 0.001). Phagocytosis assays revealed that plasmatocytes and granulocytes were able to perform phagocytosis of latex beads and L. infantum on the 1st and 2nd dpi, respectively. NO production was significantly increased (p < 0.001) on the 1st, 2nd, and 5th dpi with L. infantum and PO activity increased significantly (p < 0.05) only on the 5th dpi. L. infantum DNA was significantly increased (p < 0.001) on the 5th and 7th dpi compared to time 0. Although there are no studies describing the response of R. sanguineus s.l. to an infection with L. infantum, these results suggest that R. sanguineus s.l. activates the cellular and humoral immune response after infection with L. infantum. Further studies are however, needed to assess the impact of such a response on fighting infection.

Anatomical characterization of ultra-structures, biominerals and histolocalization of metabolites in leaves of Genipa americana

Abstract Inserted in one of the largest families among the Angiosperms, Genipa americana L., Rubiaceae, can be found in all Brazilian territory, presenting great medicinal importance, where several uses have been attributed. In view of this, this work has the purpose of analyzing the ultrastructural, biomineral, phytochemical and histochemical characteristics of the leaves of this native species from Brazil. For this, light microscopy, polarization and scanning electron microscopy techniques were used with X-ray scattering energy, associated to chromatographic and histochemical tests. The anatomical ultrastructural characteristics of the leaves detailed information about the type and arrangement of the cuticle, trichomes, surface and arrangement of the tissues that determine the botanical identity of this species. The phytochemical tests allowed determining their chromatographic pattern and histochemistry to determine the exact storage site of these substances in the leaf. It was observed that the characterization of the crystalline macro-pattern present in the analyzed species, as well as its exact elemental composition, can be considered an important differential diagnosis factor. The results characterize the leaves of this species in different aspects, being a native species and pharmacologically promising, with different popular uses and proven pharmacological activities, and more in depth studies is needed.

Macroevolution of thermal tolerance in intertidal crabs from Neotropical provinces: A phylogenetic comparative evaluation of critical limits.

Thermal tolerance underpins most biogeographical patterns in ectothermic animals. Macroevolutionary patterns of thermal limits have been historically evaluated, but a role for the phylogenetic component in physiological variation has been neglected. Three marine zoogeographical provinces are recognized throughout the Neotropical region based on mean seawater temperature (Tm): the Brazilian (Tm = 26 °C), Argentinian (Tm = 15 °C), and Magellanic (Tm = 9 °C) provinces. Microhabitat temperature (MHT) was measured, and the upper (UL 50) and lower (LL 50) critical thermal limits were established for 12 eubrachyuran crab species from intertidal zones within these three provinces. A molecular phylogenetic analysis was performed by maximum likelihood using the 16S mitochondrial gene, also considering other representative species to enable comparative evaluations. We tested for: (1) phylogenetic pattern of MHT, UL 50, and LL 50; (2) effect of zoogeographical province on the evolution of both limits; and (3) evolutionary correlation between MHT and thermal limits. MHT and UL 50 showed strong phylogenetic signal at the species level while LL 50 was unrelated to phylogeny, suggesting a more plastic evolution. Province seems to have affected the evolution of thermal tolerance, and only UL 50 was dependent on MHT. UL 50 was similar between the two northern provinces compared to the southernmost while LL 50 differed markedly among provinces. Apparently, critical limits are subject to different environmental pressures and thus manifest unique evolutionary histories. An asymmetrical macroevolutionary scenario for eubrachyuran thermal tolerance seems likely, as the critical thermal limits are differentially inherited and environmentally driven.

THE SUSCEPTIBILITY OF RECENT ISOLATES OF Schistosoma mansoni TO PRAZIQUANTEL.

INTRODUCTION: Schistosomiasis is a chronic disease caused by trematode flatworms of the genus Schistosoma and its control is dependent on a single drug, praziquantel (PZQ), but concerns over PZQ resistance have renewed interest in evaluating the in vitro susceptibility of recent isolates of Schistosoma mansoni to PZQ in comparison with well-established strains in the laboratory. MATERIAL AND METHODS: The in vitro activity of PZQ (6.5-0.003 µg/mL) was evaluated in terms of mortality, reduced motor activity and ultrastructural alterations against S. mansoni. RESULTS: After 3 h of incubation, PZQ, at 6.5 µg/mL, caused 100% mortality of all adult worms in the three types of recent isolates, while PZQ was inactive at concentrations of 0.08-0.003 µg/mL after 3 h of incubation. The results show that the SLM and Sotave isolates basically presented the same pattern of susceptibility, differing only in the concentration of 6.5 µg/mL, where deaths occurred from the range of 1.5 h in Sotave and just in the 3 h range of SLM. Additionally, this article presents ultrastructural evidence of rapid severe PZQ-induced surface membrane damage in S. mansoni after treatment with the drug, such as disintegration, sloughing, and erosion of the surface. CONCLUSION: According to these results, PZQ is very effective to induce tegument destruction of recent isolates of S. mansoni.

THE SUSCEPTIBILITY OF RECENT ISOLATES OF Schistosoma mansoni TO PRAZIQUANTEL

Introduction: Schistosomiasis is a chronic disease caused by trematode flatworms of the genus Schistosoma and its control is dependent on a single drug, praziquantel (PZQ), but concerns over PZQ resistance have renewed interest in evaluating the in vitro susceptibility of recent isolates of Schistosoma mansoni to PZQ in comparison with well-established strains in the laboratory. Material and methods: The in vitro activity of PZQ (6.5-0.003 µg/mL) was evaluated in terms of mortality, reduced motor activity and ultrastructural alterations against S. mansoni. Results: After 3 h of incubation, PZQ, at 6.5 µg/mL, caused 100% mortality of all adult worms in the three types of recent isolates, while PZQ was inactive at concentrations of 0.08-0.003 µg/mL after 3 h of incubation. The results show that the SLM and Sotave isolates basically presented the same pattern of susceptibility, differing only in the concentration of 6.5 µg/mL, where deaths occurred from the range of 1.5 h in Sotave and just in the 3 h range of SLM. Additionally, this article presents ultrastructural evidence of rapid severe PZQ-induced surface membrane damage in S. mansoni after treatment with the drug, such as disintegration, sloughing, and erosion of the surface. Conclusion: According to these results, PZQ is very effective to induce tegument destruction of recent isolates of S. mansoni.

Morphological and histological characterization of production structures, storage and distribution of venom in the parasitic wasp Bracon vulgaris.

It was described the morphology and histological composition of the structures related to production, storage and distribution of Bracon vulgaris venom, a wasp that parasite their hosts after the inoculation of a venom which causes irreversible paralysis. Were found 22 glandular filaments, coated with secretory epithelium associated with a reservoir coated internally by a chitin layer and externally by striated muscular fibers. A valve mediates the passage of the toxin to venom duct towards the parasitoids sting.

Coproduction of KPC-2 and QnrB19 in Klebsiella pneumoniae ST340 isolate in Brazil.

Few reports described the presence of bla(KPC) and qnr genes in the same isolate. This study reports the combination of bla(KPC-2) and qnrB19 genes in Klebsiella pneumoniae ST340 isolate in Brazil. These findings draw attention to this combination in ST340 isolate, which is part of the CC258, disseminated in Latin America.

Ultrastructural analysis of Leishmania infantum chagasi promastigotes forms treated in vitro with usnic acid.

Leishmaniasis is considered by the World Health Organization as one of the infectious parasitic diseases endemic of great relevance and a global public health problem. Pentavalent antimonials used for treatment of this disease are limited and new phytochemicals emerge as an alternative to existing treatments, due to the low toxicity and cost reduction. Usnic acid is uniquely found in lichens and is especially abundant in genera such as Alectoria, Cladonia, Evernia, Lecanora, Ramalina, and Usnea. Usnic acid has been shown to exhibit antiviral, antiprotozoal, antiproliferative, anti-inflammatory, and analgesic activity. The aim of this study was to evaluate the antileishmanial activity of usnic acid on Leishmania infantum chagasi promastigotes and the occurrence of drug-induced ultrastructural damage in the parasite. Usnic acid was effective against the promastigote forms (IC50=18.30±2.00 µg/mL). Structural and ultrastructural aspects of parasite were analyzed. Morphological alterations were observed as blebs in cell membrane and shapes given off, increasing the number of cytoplasmic vacuoles, and cellular and mitochondrial swelling, with loss of cell polarity. We concluded that the usnic acid presented antileishmanial activity against promastigote forms of Leishmania infantum chagasi and structural and ultrastructural analysis reinforces its cytotoxicity. Further, in vitro studies are warranted to further evaluate this potential.

Simplagrin, a platelet aggregation inhibitor from Simulium nigrimanum salivary glands specifically binds to the Von Willebrand factor receptor in collagen and inhibits carotid thrombus formation in vivo.

BACKGROUND: Among the several challenges faced by bloodsucking arthropods, the vertebrate hemostatic response against blood loss represents an important barrier to efficient blood feeding. Here we report the first inhibitor of collagen-induced platelet aggregation derived from the salivary glands of a black fly (Simulium nigrimanum), named Simplagrin. METHODS AND FINDINGS: Simplagrin was expressed in mammalian cells and purified by affinity-and size-exclusion chromatography. Light-scattering studies showed that Simplagrin has an elongated monomeric form with a hydrodynamic radius of 5.6 nm. Simplagrin binds to collagen (type I-VI) with high affinity (2-15 nM), and this interaction does not involve any significant conformational change as determined by circular dichroism spectroscopy. Simplagrin-collagen interaction is both entropically and enthalpically driven with a large negative ΔG, indicating that this interaction is favorable and occurs spontaneously. Simplagrin specifically inhibits von Willebrand factor interaction with collagen type III and completely blocks platelet adhesion to collagen under flow conditions at high shear rates; however, Simplagrin failed to block glycoprotein VI and Iα2ß1 interaction to collagen. Simplagrin binds to RGQOGVMGF peptide with an affinity (K(D) 11 nM) similar to that of Simplagrin for collagen. Furthermore, Simplagrin prevents laser-induced carotid thrombus formation in vivo without significant bleeding in mice and could be useful as an antithrombotic agent in thrombosis related disease. CONCLUSION: Our results support the orthology of the Aegyptin clade in bloodsucking Nematocera and the hypothesis of a faster evolutionary rate of salivary function of proteins from blood feeding arthropods.

Ultrastructural aspects of melatonin cytotoxicity on Caco-2 cells in vitro.

Colon adenocarcinoma is a disease expanding worldwide. Cancer of colon and rectum are among the top ten most insidious types in Brazil. In vitro and in vivo studies have demonstrated the efficacy of the hormone melatonin to prevent and reduce tumor growth. However, there are only few studies addressing the action of melatonin on Caco-2 cells. Thus, the cytotoxic effect of melatonin on the ultrastructure of Caco-2 cells was investigated. The MTT colorimetric method was used to assess the cytotoxicity. A total of 2×10(6)cells/mL were seeded in microplates and incubated at 50, 25, 12.5, 6.25, 3.125, 1.56, 0.78 and 0.0 (control) µg/mL of melatonin. For ultrastructural analysis concentrations with low, medium and high cytotoxicity plus the control were used for ultrastructural analysis. The concentrations 50, 1.56 and 0.78 µg/mL of melatonin showed low, medium and high cytotoxicity, respectively. Ultrastructurally, the control tumor cells were shown to be preserved. Caco-2 cells showed morphological changes at 50 µg/mL of melatonin, with numerous vacuoles, mitochondrial degeneration and reduced glycogen. However, Caco-2 cells also showed altered morphology in treatments at 1.56 and 0.78 µg/mL of melatonin with characteristics of cells in degeneration by the presence of numerous vacuoles, absence of microvilli, mitochondrial degeneration and nuclear fragmentation. Thus, one can infer that concentrations of 1.56 and 0.78 µg/mL of melatonin promote cytotoxicity in Caco-2 cells, which can probably be related to the generation of reactive oxygen species (ROS).

Perception of local inhabitants regarding the socioeconomic impact of tourism focused on provisioning wild dolphins in Novo Airão, Central Amazon, Brazil.

Botos (Inia geoffrensis) are currently provisioned for use in tourist attractions in five sites in the Brazilian Amazon. Despite the known negative effects associated with human-wild dolphin interactions, this activity has been regulated and licensed in the Anavilhanas National Park in Novo Airão, Amazonas State, Brazil. We present an updated evaluation of the perception of the local community concerning the possible socioeconomic impacts of this tourism in Novo Airão. In April 2011, 45 interviews were conducted with inhabitants. A small segment of Novo Airão perceives currently itself as being economically dependent on the botos feeding tourism. Despite that, the economic benefits of this controversial activity apparently are not shared among most inhabitants, and botos feeding tourism is perceived as generating diverse negative effects. We conclude that if the activity was banned or modified into a less impacting tourist activity, this action would probably not majorly affect the lives of the general population.

Microscopic analysis of Spodoptera frugiperda (Lepidoptera: Noctuidae) embryonic development before and after treatment with azadirachtin, lufenuron, and deltamethrin.

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.