The Role of DmCatD, a Cathepsin D-Like Peptidase, and Acid Phosphatase in the Process of Follicular Atresia in Dipetalogaster maxima (Hemiptera: Reduviidae), a Vector of Chagas' Disease

In this work, we have investigated the involvement of DmCatD, a cathepsin D-like peptidase, and acid phosphatase in the process of follicular atresia of Dipetalogaster maxima, a hematophagous insect vector of Chagas' disease. For the studies, fat bodies, ovaries and hemolymph were sampled from anautogenous females at representative days of the reproductive cycle: pre-vitellogenesis, vitellogenesis as well as early and late atresia. Real time PCR (qPCR) and western blot assays showed that DmCatD was expressed in fat bodies and ovaries at all reproductive stages, being the expression of its active form significantly higher at the atretic stages. In hemolymph samples, only the immunoreactive band compatible with pro-DmCatD was observed by western blot. Acid phosphatase activity in ovarian tissues significantly increased during follicular atresia in comparison to pre-vitellogenesis and vitellogenesis. A further enzyme characterization with inhibitors showed that the high levels of acid phosphatase activity in atretic ovaries corresponded mainly to a tyrosine phosphatase. Immunofluorescence assays demonstrated that DmCatD and tyrosine phosphatase were associated with yolk bodies in vitellogenic follicles, while in atretic stages they displayed a different cellular distribution. DmCatD and tyrosine phosphatase partially co-localized with vitellin. Moreover, their interaction was supported by FRET analysis. In vitro assays using homogenates of atretic ovaries as the enzyme source and enzyme inhibitors demonstrated that DmCatD, together with a tyrosine phosphatase, were necessary to promote the degradation of vitellin. Taken together, the results strongly suggested that both acid hydrolases play a central role in early vitellin proteolysis during the process of follicular atresia.Fil: Leyria, Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Fruttero, Leonardo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Nazar, Magalí. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Canavoso, Lilian Etelvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentin

DmCatD, a cathepsin D-like peptidase of the hematophagous insect Dipetalogaster maxima (Hemiptera: Reduviidae): Purification, bioinformatic analyses and the significance of its interaction with lipophorin in the internalization by developing oocytes

DmCatD, a cathepsin D-like peptidase of the hematophagous insect Dipetalogaster maxima, is synthesized by the fat body and the ovary and functions as yolk protein precursor. Functionally, DmCatD is involved in vitellin proteolysis. In this work, we purified and sequenced DmCatD, performed bioinformatic analyses and investigated the events involved in its targeting and storage in developing oocytes. By ion exchange and gel filtration chromatography, DmCatD was purified from egg homogenates and its identity was confirmed by mass spectrometry. Approximately 73% of the full-length transcript was sequenced. The phylogeny indicated that DmCatD has features which suggest its distancing from “classical” cathepsins D. Bioinformatic analyses using a chimeric construct were employed to predict post-translational modifications. Structural modeling showed that DmCatD exhibited the expected folding for this type of enzyme, and an active site with conserved architecture. The interaction between DmCatD and lipophorin in the hemolymph was demonstrated by co-immunoprecipitation. Colocalization of both proteins in developing oocyte membranes and yolk bodies was detected by immunofluorescence. Docking assays favoring the interaction DmCatD-lipophorin were carried out after modeling lipophorin of a related triatomine species. Our results suggest that lipophorin acts as a carrier for DmCatD to facilitate its further internalization by the oocytes. The mechanisms involved in the uptake of peptidases within the oocytes of insects have not been reported. This is the first experimental work supporting the interaction between cathepsin D and lipophorin in an insect species, enabling us to propose a pathway for its targeting and storage in developing oocytes.Fil: Leyria, Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; ArgentinaFil: Fruttero, Leonardo Luis. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentina. Pontificia Universidade Católica do Rio Grande do Sul; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Ligabue Braun, Rodrigo. Universidade Federal do Rio Grande do Sul; BrasilFil: Defferrari, Marina S.. University of Toronto; CanadáFil: Arrese, Estela L.. Oklahoma State University; Estados UnidosFil: Soulages, José L.. Oklahoma State University; Estados UnidosFil: Settembrini, Beatriz Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales “Bernardino Rivadavia”; ArgentinaFil: Carlini, Célia Regina R S. Pontificia Universidade Católica do Rio Grande do Sul; BrasilFil: Canavoso, Lilian Etelvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentin

Morphological and ultrastructural characterization of hemocytes in an insect model, the hematophagous dipetalogaster maxima (Hemiptera: Reduviidae)

Hemocytes, the cells present in the hemolymph of insects and other invertebrates, perform several physiological functions, including innate immunity. The current classification of hemocyte types is based mostly on morphological features; however, divergences have emerged among special-ists in triatomines, the insect vectors of Chagas’ disease (Hemiptera: Reduviidae). Here, we have combined technical approaches in order to characterize the hemocytes from fifth instar nymphs of the triatomine Dipetalogaster maxima. Moreover, in this work we describe, for the first time, the ultrastructural features of D. maxima hemocytes. Using phase contrast microscopy of fresh prepara-tions, five hemocyte populations were identified and further characterized by immunofluorescence, flow cytometry and transmission electron microscopy. The plasmatocytes and the granulocytes were the most abundant cell types, although prohemocytes, adipohemocytes and oenocytes were also found. This work sheds light on a controversial aspect of triatomine cell biology and physiology setting the basis for future in-depth studies directed to address hemocyte classification using non-microscopy-based markers.Fil: Moyetta, Natalia Rita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Ramos, Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Leyria, Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Canavoso, Lilian Etelvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Fruttero, Leonardo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentin

Cell death mechanisms during follicular atresia in Dipetalogaster maxima, a vector of Chagas' disease (Hemiptera: Reduviidae)

In this work wehave analyzed the involvement of cell death pathways during the process of follicular atresia in the hematophagous insect vector Dipetalogaster maxima. Standardized insect rearing conditions were established to induce agradual follicular degeneration stage bydepriving females ofblood meal during post-vitellogenesis. Wefirstcharacterized the morpho-histological and ultrastructural changes of the ovarian tissue at early and late follicular atresia by light and transmission electron microscopy. Apoptosis was investigated by DAPI nuclear staining, TUNEL labeling and the detection ofactive caspase-3 by immunofluorescence. Autophagy was assessed bythe measurement ofacid phosphatase activ- ity in ovarian homogenates and monitor edbythe detection ofthe specificmarker ofautophagic compartments, LC3. High levels of acid phosphatase activity were detected at all atretic stages. However, follicular cells of follicles undergoing incipient degeneration in early atresia exhibited features of apoptosis such as chromatin condensation,DNA fragmentation and the presence of active caspase-3. The ultrastructural findings and the increased levels of LC3-II found at late follicular atresia supported the relevance of autophagy at this atretic stage, although the extent of autophagosome formation demonstrated that this cell death pathway also occurred at early atresia. In late atresia,follicular cells also displayed more drastic changes compatible with necrosis. Taken together, results showed that apoptosis, autophagy and necrosis were operative during follicular atresia in D. maxima. Moreover, it was shown that the relevance of these cell death mechanisms correlate swith the time elapsed since the onset of the degenerative process.Fil: Aguirre, Silvina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Pons, Patricia. Universidad Nacional de Córdoba. Facultad de Medicina. Centro de Microscopía Electrónica; ArgentinaFil: Settembrini, Beatriz Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Arroyo, Daniela Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Canavoso, Lilian Etelvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentin

The fat body of the hematophagous insect, panstrongylus megistus (Hemiptera: Reduviidae): Histological features and participation of the β-chain of ATP synthase in the lipophorin-mediated lipid transfer

In insects, lipid transfer to the tissues is mediated by lipophorin, the major circulating lipoprotein, mainly through a nonendocytic pathway involving docking receptors. Currently, the role of such receptors in lipid metabolism remains poorly understood. In this work, we performed a histological characterization of the fat body of the Chagas’ disease vector, Panstrongylus megistus (Burmeister), subjected to different nutritional conditions. In addition, we addressed the role of the β-chain of ATP synthase (β-ATPase) in the process of lipid transfer from lipophorin to the fat body. Fifth-instar nymphs in either fasting or fed condition were employed in the assays. Histological examination revealed that the fat body was composed by diverse trophocyte phenotypes. In the fasting condition, the cells were smaller and presented a homogeneous cytoplasmic content.The fat body of fed insects increased in size mainly due to the enlargement of lipid stores. In this condition, trophocytes contained abundant lipid droplets, and the rough endoplasmic reticulum was highly developed and mitochondria appeared elongated. Immunofluorescence assays showed that the β-ATPase, a putative lipophorin receptor, was located on the surface of fat body cells colocalizing partially with lipophorin, which suggests their interaction. No changes in β-ATPase expression were found in fasting and fed insects. Blocking the lipophorin–β-ATPase interaction impaired the lipophorin-mediated lipid transfer to the fat body. The results showed that the nutritional status of the insect influenced the morphohistological features of the tissue. Besides, these findings suggest that β-ATPase functions as a lipophorin docking receptor in the fat body.Fil: Fruttero, Leonardo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Leyria, Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Moyetta, Natalia Rita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Ramos, Fabián O.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Settembrini, Beatriz Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Canavoso, Lilian Etelvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentin

Host influence on the nutritional and reproductive status of Triatoma infestans (Klug) (Hemiptera: Reduviidae) peridomiciliary populations

Triatoma infestans is the main vector of Chagas disease in the southern cone of South America. This species is well adapted to living in rural houses and structures used for housing domestic animals (peridomestic habitats). In this study, we evaluated the relationship between the source of blood consumed by adults of T. infestans collected from different peridomestic habitats from two localities from Cruz del Eje department (Córdoba, Argentina) and their nutritional and reproductive status. In each individual, the ratio between body weight and total body length was used as an indicator of nutritional status (NS). The presence of sperm in spermathecae and the number of chorionated oocytes in ovaries and oviducts were considered indicators of reproductive status (RS) of females. The feeding source in the promesenteron of male and female insects was identified using anti-chicken, anti-goat, antihuman and anti-dog antisera. Chicken coops were the main peridomestic structure present in the study area as well as the peridomestic sites with the highest percentage of T. infestans. Insects collected from the different peridomestic structures showed a NS between 8 and 15 mg/mm. Of the evaluated females, 35.7% presented chorionated oocytes. Food profile analyses revealed that chicken was the main blood source. Independently of the blood source, the triatomines presented a NS between 8 and 15 mg/mm. No specimens feeding exclusively on human blood were found; nevertheless, of 31.48% of insects feeding on mixed blood sources, 59% included human blood. All T. infestans specimens that included human blood in the mixed blood source were collected from chicken coops and storerooms located in a 12-m area around domiciles. Human blood present in mixed blood meal of adult insects suggests that T. infestans moves from domiciles to peridomicilies and vice versa.Triatoma infestans es el principal vector de la enfermedad de Chagas en el Cono Sur de Sudamérica. Esta especie está bien adaptada a vivir en el domicilio y estructuras utilizadas para albergar animales domésticos (hábitats peridomésticos). En este trabajo evaluamos la relación entre la fuente de sangre consumida por los adultos de T. infestans recolectados de diferentes hábitats peridomésticos de dos localidades del departamento de Cruz del Eje (Córdoba, Argentina) y su estado nutricional y reproductivo. En cada individuo, la relación entre el peso y la longitud corporal total se utilizó como un indicador del estado nutricional (EN). La presencia de espermatozoides en espermatecas y el número de ovocitos corionados en ovarios y oviductos se consideraron indicadores del estado reproductivo (ER) de las hembras. La fuente de alimentación en el promesenterón de insectos machos y hembras se identificó utilizando antisueros anti-gallina, anti-cabra, anti-humano y anti-perro. Los gallineros fueron las principales estructuras peridomésticas presentes en el área de estudio, así como los sitios con el mayor porcentaje de T. infestans. Los insectos recolectados en las diferentes estructuras peridomésticas mostraron un EN entre 8 y 15 mg / mm. De las hembras evaluadas, el 35,7% presentó ovocitos corionados. Los análisis del perfil alimentario revelaron que las gallinas fueron la principal fuente de sangre. Independiente de la fuente de sangre los triatominos presentaron EN entre 8 y 15 mg / mm. No se encontraron ejemplares alimentados exclusivamente con sangre humana; sin embargo, del 31,48% de los insectos que se alimentaron de fuentes de sangre mixtas, el 59% incluía sangre humana. Todas las muestras de T. infestans que incluían sangre humana en la fuente de sangre se recolectaron en gallineros y depósitos ubicados en un área de 12 m alrededor de los domicilios. La sangre humana presente en las fuentes de alimentación mixta sugiere que T. infestans se mueve de los domicilios a los peridomicilios y viceversa.Fil: Soria, Carola. Universidad Nacional de Córdoba. Facultad de Cs.exactas Físicas y Naturales. Departamento de Fisiología. Cátedra de Introducción A la Biología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Cardozo, Miriam. Universidad Nacional de Córdoba. Facultad de Cs.exactas Físicas y Naturales. Departamento de Fisiología. Cátedra de Introducción A la Biología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Canavoso, Lilian Etelvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Crocco, Liliana Beatriz. Universidad Nacional de Córdoba. Facultad de Cs.exactas Físicas y Naturales. Departamento de Fisiología. Cátedra de Introducción A la Biología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Nattero, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución. Laboratorio de Eco-Epidemiología; ArgentinaFil: Ortiz, Valeria. Universidad Nacional de Córdoba. Facultad de Cs.exactas Físicas y Naturales. Departamento de Fisiología. Cátedra de Introducción A la Biología; ArgentinaFil: Leyria, Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Rodriguez, Claudia Susana. Universidad Nacional de Córdoba. Facultad de Cs.exactas Físicas y Naturales. Departamento de Fisiología. Cátedra de Introducción A la Biología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentin

The in vivo treatment with the plant urease “Jack Bean Urease” impaired reproduction in females of Rhodnius prolixus (Hemiptera: Reduviidae)

Ureases are enzymes that catalyze the hydrolysis of urea to carbon dioxide and ammonia. In recent decades, it has been postulated that plant ureases are also defense proteins against phytophagous insect species with potential biotechnological application. Previous reports demonstrated that the injection of "Jack Bean Urease" (JBU), the major isoform of urease from the legume Canavalia ensiformis, into the hemocele of triatomine insects, resulted in several toxic effects including activation of the immune response. Although the insecticidal effect of JBU was described several years ago, many aspects of its mechanism of action as well as the target organs remains largely uncharacterized. In particular, the effects of JBU on the female reproductive system and the consequences of sublethal doses have not been studied.Para acceder a la videoconferencia completa, hacer clic en "Enlace externo".Sociedad Latinoamericana de Ecología de Vectore

Patrón de desarrollo ovárico en hembras de Triatoma sordida ayunadas

Entre los vectores secundarios de Trypanosoma cruzi - agente causante de enfermedad de chagas - Triatoma sordida es considerada una especie de importancia epidemiológica debido a su presencia en peridomicilios y a la tendencia a invadir y formar grandes colonias domésticas.Fil: Rodríguez, C. Universidad Nacional de Córdoba Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Biológicas y Tecnológicas. Cátedra Introducción a la Biología; Argentina.Fil: Rodríguez, C. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales; Argentina.Fil: Aguirre, S. Universidad Nacional de Córdoba. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones en Bioquímica Clínica e Inmunología. Departamento de Bioquímica Clínica; Argentina.Fil: Aguirre, S. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas; Argentina.Fil: Nattero, J. Universidad Nacional de Córdoba Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Biológicas y Tecnológicas. Cátedra .Introducción a la Biología; Argentina.Fil: Nattero, J. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales; Argentina.Fil: Canavoso, L. Universidad Nacional de Córdoba. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones en Bioquímica Clínica e Inmunología. Departamento de Bioquímica Clínica; Argentina.Fil: Canavoso, L. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas; Argentina.Fil: Crocco, L. Universidad Nacional de Córdoba Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Biológicas y Tecnológicas. Cátedra .Introducción a la Biología; Argentina.Fil: Crocco, L. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales; Argentina.Zoología, Ornitología, Entomología, Etologí

Flight behaviour of European corn borer infected with Nosema pyrausta

The microsporidian Nosema pyrausta Paillot is a common and widespread pathogen of European corn borer, Ostrinia nubilalis (Hübner), in North America and Europe. Nosema negatively affects European corn borer longevity and fecundity. In this study, we used flight mills to examine the effects of Nosema infection on flight activity of unmated males and females at 1 and 3 days after pupal eclosion, taking the level of infection, as measured by number of Nosema spores per mg of tissue, into consideration. Nosema infection had a significant negative effect on distance, duration and speed of the longest uninterrupted flight, as well as on total distance and duration of flight of 1-day-old males, but not of 3-day-old males or females of either age. However, when insects with a light infection (\u3c15 spores/mg) were pooled with uninfected moths, significant negative effects of a moderate/heavy infection (≥15 spores/mg) were observed for most flight parameters in 1-day-old, but not 3-day-old, moths of both sexes. The magnitude of reduction was often substantial: e.g., distance and duration were sevenfold, and 3.5-fold less, respectively, in 1-day-old females with a moderate/heavy infection. Flight distance and duration were significantly negatively correlated with level of Nosema infection in 1-day-old, but not 3-day-old, moths of both sexes. The percentage of moths of either sex or age with a moderate/heavy Nosema infection flying given distances was less than that of lightly infected (\u3c15 spores/mg) moths. Among uninfected adults, forewing length was positively correlated with several measures of flight performance in males, but not females. However, some performance measures were correlated with wing length in females when infected with Nosema. Nosema infection was associated with reduced adult weight, but not forewing length, suggesting that negative effects of Nosema infection on flight performance are related to reduced energy reserves rather than smaller body size per se

Apolipophorin-III Mediates Antiplasmodial Epithelial Responses in Anopheles gambiae (G3) Mosquitoes

Apolipophorin-III (ApoLp-III) is known to play an important role in lipid transport and innate immunity in lepidopteran insects. However, there is no evidence of involvement of ApoLp-IIIs in the immune responses of dipteran insects such as Drosophila and mosquitoes.We report the molecular and functional characterization of An. gambiae apolipophorin-III (AgApoLp-III). Mosquito ApoLp-IIIs have diverged extensively from those of lepidopteran insects; however, the predicted tertiary structure of AgApoLp-III is similar to that of Manduca sexta (tobacco hornworm). We found that AgApoLp-III mRNA expression is strongly induced in the midgut of An. gambiae (G3 strain) mosquitoes in response to Plasmodium berghei infection. Furthermore, immunofluorescence stainings revealed that high levels of AgApoLp-III protein accumulate in the cytoplasm of Plasmodium-invaded cells and AgApoLp-III silencing increases the intensity of P. berghei infection by five fold.There are broad differences in the midgut epithelial responses to Plasmodium invasion between An. gambiae strains. In the G3 strain of An. gambiae AgApoLp-III participates in midgut epithelial defense responses that limit Plasmodium infection