Derris (Lonchocarpus) urucu (Leguminosae) Extract Modifies the Peritrophic Matrix Structure of Aedes aegypti (Diptera:Culicidae)

Aqueous suspension of ethanol extracts of Derris (Lonchocarpus) urucu   (Leguminosae), collected in the state of Amazonas, Brazil, were tested for larvicidal activity against the mosquito Aedes aegypti (Diptera:Culicidae). The aim of this study was to observe the alterations of peritrophic matrix in Ae. aegypti larvae treated with an aqueous suspension of D. urucu extract. Different concentrations of D. urucu root extract were tested against fourth instar larvae. One hundred percent mortality was observed at 150 μg/ml (LC50 17.6 μg/ml) 24 h following treatment. In response to D. urucu feeding, larvae excreted a large amount of amorphous feces, while control larvae did not produce feces during the assay period. Ultrastructural studies showed that larvae fed with 150 μg/ml of D. urucu extract for 4 h have an imperfect peritrophic matrix and extensive damage of the midgut epithelium. Data indicate a protective role for the peritrophic matrix. The structural modification of the peritrophic matrix is intrinsically associated with larval mortality

First isolation of microorganisms from the gut diverticulum of Aedes aegypti (Diptera: Culicidae): New perspectives for an insect-bacteria association

We show for the first time that the ventral diverticulum of the mosquito gut (impermeable sugar storage organ) harbors microorganisms. The gut diverticulum from newly emerged and non-fed Aedes aegypti was dissected under aseptic conditions, homogenized and plated on BHI medium. Microbial isolates were identified by sequencing of 16S rDNA for bacteria and 28S rDNA for yeast. A direct DNA extraction from Ae. aegypti gut diverticulum was also performed. The bacterial isolates were: Bacillus sp., Bacillus subtilis and Serratia sp. The latter was the predominant bacteria found in our isolations. The yeast species identified was Pichia caribbica

First isolation of microorganisms from the gut diverticulum of Aedes aegypti (Diptera: Culicidae): new perspectives for an insect-bacteria association

We show for the first time that the ventral diverticulum of the mosquito gut (impermeable sugar storage organ) harbors microorganisms. The gut diverticulum from newly emerged and non-fed Aedes aegypti was dissected under aseptic conditions, homogenized and plated on BHI medium. Microbial isolates were identified by sequencing of 16S rDNA for bacteria and 28S rDNA for yeast. A direct DNA extraction from Ae. aegypti gut diverticulum was also performed. The bacterial isolates were: Bacillus sp., Bacillus subtilis and Serratia sp. The latter was the predominant bacteria found in our isolations. The yeast species identified was Pichia caribbica

Polyphenol-Rich Diets Exacerbate AMPK-Mediated Autophagy, Decreasing Proliferation of Mosquito Midgut Microbiota, and Extending Vector Lifespan

Submitted by Sandra Infurna ([email protected]) on 2017-03-16T12:33:29Z No. of bitstreams: 1 rubem2_mennabarreto_etal_IOC_2016.pdf: 2327344 bytes, checksum: 6800e46b47583d5b57ed1a97317d8d30 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2017-03-16T12:57:48Z (GMT) No. of bitstreams: 1 rubem2_mennabarreto_etal_IOC_2016.pdf: 2327344 bytes, checksum: 6800e46b47583d5b57ed1a97317d8d30 (MD5)Made available in DSpace on 2017-03-16T12:57:48Z (GMT). No. of bitstreams: 1 rubem2_mennabarreto_etal_IOC_2016.pdf: 2327344 bytes, checksum: 6800e46b47583d5b57ed1a97317d8d30 (MD5) Previous issue date: 2016Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Sinalização Celular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Sinalização Celular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Sinalização Celular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Sinalização Celular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Sinalização Celular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Sinalização Celular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil / Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Bioquímica de Lipídios e Lipoproteínas. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Sinalização Celular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Sinalização Celular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Celular. Rio de Janeiro, RJ. Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Bioquímica de Artrópodes Hematófagos. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Instituto Federal de Educacão, Ciência e Tecnologia Fluminense. Laboratório de Biologia. Campos dos Goytacazes, RJ, Brasil.Universidade Estadual do Norte Fluminense. Laboratório de Biotecnologia. Campos dos Goytacazes, RJ, Brasil.Universidade Federal do Rio de Janeiro. Centro de Ciências da Saúde. Instituto de Bioquímica Médica Leopoldo De Meis. Rio de Janeiro, RJ, Brasil.Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil / Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Bioquímica de Artrópodes Hematófagos. Rio de Janeiro, RJ, Brasil.Simon Fraser University. Department of Biological Sciences. Burnaby, British Columbia, Canada.Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil / Universidade Federal do Rio de Janeiro. Faculdade de Farmácia. Departamento de Biotecnologia Farmacêutica. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Sinalização Celular. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil / Universidade Federal do Rio de Janeiro. Instituto de Química. Departamento de Bioqúímica. Laboratório de Bioinformática. Rio de Janeiro, RJ, Brasil.Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular. Rio de Janeiro, RJ, Brasil / Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Bioquímica de Lipídios e Lipoproteínas. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Sinalização Celular. Rio de Janeiro, RJ, Brasil / Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo De Meis. Programa de Biologia Molecular e Biotecnologia. Laboratório de Bioquímica de Lipídios e Lipoproteínas. Rio de Janeiro, RJ, Brasil.Mosquitoes feed on plant-derived fluids such as nectar and sap and are exposed to bioactive molecules found in this dietary source. However, the role of such molecules on mosquito vectorial capacity is unknown. Weather has been recognized as a major determinant of the spread of dengue, and plants under abiotic stress increase their production of polyphenols

Rv treatment downregulates apoptotic pathway.

<p>Adult mosquitoes were reared until six days old under each dietary condition. The midguts of female mosquitoes were dissected and homogenized in TRIzol, and total RNA was extracted. These samples were used to perform qPCR for the genes: (<b>A</b>) 16S, (<b>B</b>) ARGONAUTE 2, (<b>C</b>) CASPASE 16, (<b>D</b>) AeDRONC, (<b>E</b>) AeIAP1 and (<b>F</b>) ATG8. Data show means and standard error of at least three independent experiments. Ctrl—0.05% ethanol plus 10% sucrose; Rv—100 μM Rv in 0.05% ethanol plus 10% sucrose; AbMix– 10% sucrose, 10 U/mL penicillin, 10 U/mL streptomycin and 15 U/mL gentamicin. *—p<0.05; **—p<0.01; ***—p<0.001; ****—p<0.0001, as calculated by Student’s t-test (F as calculated by one-way ANOVA with Tukey post test). (Error bars, s.e.m., n = 3 experiments).</p

Summary data of the effects of polyphenols on mosquito lifespan.

<p>Summary data of the effects of polyphenols on mosquito lifespan.</p

Rv triggers autophagy in mosquito midgut.

<p><b>(A, B</b>) Typical ultrastructural appearance of mosquito midgut fed on a control diet. (<b>C-F</b>) Midguts from insects fed on Rv. White stars indicate the formation of concentric membrane structures. Black arrows indicate preserved mitochondrial morphology and normal cristae aspects. N: nucleus; M: mitochondria; Mi: microvilli. Bars in panels A and C: 2 μm. Bars in panels B, D-F: 0.5 μm. (<b>G</b>) Midgut images obtained under a fluorescence microscope after incubation with LysoTracker Red (upper panels, DIC) (lower panels, fluorescence). (<b>H</b>) Densitometry of LysoTracker fluorescence images shown in panel G. (<b>I)</b> Images obtained as in panel G following the silencing of mosquito AMPK (DsAMPK) or an unrelated protein (DsMal). Insects were kept on the indicated Control (Ctrl), Rv or AICAR diets. <b>(J</b>) Densitometry of LysoTracker fluorescence images shown in the lower part of panels I. Rv—100 μM Rv; AICAR—1 mM AICAR. **—p<0.01; ****—p<0.0001 as calculated by one-way ANOVA with Tukey post test (I) and by Student’s t-test (J). Data in panels I, J represent means and s. e. m. (n = 3).</p

Polyphenols increase mosquito lifespan while still allowing dengue virus infection.

<p>(<b>A-D</b>) Effect of polyphenols on male (<b>A, C</b>) and female (<b>B, D</b>) lifespan. The 50% mean lifespan is indicated by the horizontal dotted line. (<b>E</b>) Dengue virus RNA from mosquitoes fed or not before infection with Rv. (<b>F</b>) Percentage of dengue-infected mosquitoes. Ctrl—Control, Epi—100 μM epi-gallo-catechin-gallate; Gen—100 μM genistein; Que—100 μM quercetin. (Error bars,s.e.m., n = 3 experiments with 200 mosquitoes in each cage, p<0.05 in each survivor curve compared with control as calculated by Mantel-Cox and Gehan-Breslow-Wilcoxon test).</p

Dietary polyphenols decrease lipid accumulation in mosquitoes through AMPK activation.

<p>(<b>A</b>) Densitometric analysis of triacylglycerol content measured by TLC. (<b>B</b>) Densitometric analysis of western blots for midgut p-AMPK. (<b>C</b>) Densitometric analysis of western blots for fat body p-AMPK. (<b>D</b>) Densitometric analysis of triacylglycerol levels as measured by TLC in mosquitoes fed as indicated. Actin was used as loading control in panels B and C. (HepG2) HepG2 cells are a positive control for pAMPK antibody. Insets show representative images of either TLCs or blots. Ctrl—control; Rv—100 μM Rv; Epi—100 μM epi-gallo-catechin-gallate; Gen—100 μM genistein; Que—100 μM quercetin; CC—100 μM Compound C; CC+Rv—100 μM Compound C and 100 μM Rv; AICAR—1 mM AICAR. *—p<0.05; **—p<0.01, as calculated by one-way ANOVA with Tukey post test. (Error bars, s.e.m., n = 3 experiments).</p