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

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

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

RESEARCH NOTE - Adipokinetic Hormone Causes Formation of Low Density Lipophorin in the Hemolymph of Triatoma infestans

The importance of the presence of lipids in insects has been taken into account for many years. The lipids travel throughout the hemolymph from the midgut to the fat body and other organs, and this also occurs in mammals in a similar way; they associate with proteins transforming into the hemolymphatic lipoproteins or lipophorins (H Chino &amp: KJ Kitazawa 1981 J Lipid Res 22: 1042-1051, AM Beenakkers et al. 1985 Prog Lipid Res 24: 19-67)

Molecular response of Musca domestica L. to Mintostachys verticillata essential oil, (4R)(+)-pulegone and menthone

Intense applications of synthetic insecticides for the control of adult Musca domestica have led to the insects developing resistance to most of them. In consequence, there is interest in new active ingredients as alternatives to conventional insecticides. Essential oils (EO) are potential tools for controlling M. domestica because of their effectiveness and their minimal environmental effects. In a fumigant assay, M. domestica adults treated with Minthostachys verticillata EO [LC50 = 0.5 mg/dm3; majority components by SPME-GC: (4R)(+)-pulegone (67.5%), menthone (22.3%) and (4R)(+)-limonene (3.8%)], died within 15 min or less. The terpenes absorbed by the flies and their metabolites, analyzed using SPME fiber, were (4R)(+)-limonene (LC50 = 6.2 mg/dm3), menthone (LC 50 = 1.9 mg/dm3), (4R)(+)-pulegone (LC50 = 1.7 mg/dm3) and a new component, menthofuran (LC50 = 0.3 mg/dm3), in a relative proportion of 12.4, 6.5, 35.9 and 44.2% respectively. Menthofuran was formed by oxidation of either (4R)(+)-pulegone or menthone mediated by cytochrome P450, as demonstrated by a fumigation assay on flies previously treated with piperonyl butoxide, a P450 inhibitor, which showed a decrease in toxicity of the EO, (4R)(+)-pulegone and of menthone, supporting the participation of the P450 oxidizing system in the formation of menthofuran. The enzymatic reaction of isolated fly microsomes with the EO or the (4R)(+)-pulegone produced menthofuran in both cases. Contrary to expectations, the insect detoxification system contributed to enhance the toxicity of the M. verticillata EO. Consequently, resistant strains overexpressing P450 genes will be more susceptible to either M. verticillata EO or (4R)(+)-pulegone and menthone.Fil: Rossi, Yanina Estefanía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones y Transferencia de Villa María. Universidad Nacional de Villa María. Centro de Investigaciones y Transferencia de Villa María; ArgentinaFil: Canavoso, Lilian Etelvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Palacios, Sara Maria. Universidad Católica de Córdoba. Facultad de Ciencias Químicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Fat body morphology, but not body size, changes in forager bees of Scaptotrigona jujuyensis (Apidae: Meliponini) during foraging season

The morphological variation of workers is a phenomenon related to a colony context. We have studied body size and fat body morphology in foragers of Scaptotrigona jujuyensis during different periods of the foraging season. In workers which forage at the start of season the number of oenocytes was lower than that of the remaining groups. Cytological analyses of trophocytes revealed that the granules differed in the staining pattern according to the period of sampling. The differences in the fat body of the workers may be due to the complexity and dynamic turnover of the vacuolar system of the trophocytes. Our results suggest that fat body morphology in adults of Scaptotrigona jujuyensis changes during foraging. The fat body might play a key role as an organ contributing to the success of a colony through its plastic changes in worker bees.Fil: Galvani, Gerónimo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Soto, Eduardo Maria. 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; ArgentinaFil: Canavoso, Lilian Etelvina. Universidad Nacional de Córdoba; Argentina. 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"; Argentin