Anopheles darlingi Root 1926 (Diptera: Culicidae): variaciones morfométricas en alas y patas de poblaciones de Colombia / Anopheles darlingi (Diptera: Culicidae) Rood 1926: Morphometric variations in wings and legs of populations from Colombia

Resumen Introducción. Las poblaciones naturales de Anopheles darlingi, principal transmisor de la malaria en Colombia, han mostrado plasticidad fenotípica en algunos de sus caracteres diagnósticos. Objetivo. Caracterizar las variaciones morfométricas de los patrones alares y de la pata posterior en poblaciones naturales de An. darlingi recolectados en localidades colombianas donde la malaria es endémica. Materiales y métodos. Se analizaron los patrones de manchas de la vena costal del ala de hembras silvestres recolectadas en los departamentos de Chocó, Guaviare, Meta y Vichada mediante morfometría lineal y geométrica. El segundo tarsómero posterior de las hembras se analizó mediante morfometría lineal. Resultados. Se encontraron 19 patrones de manchas de la vena costal. Los patrones I, con 49 % (n=118/240) y VI, con 28 % (n=66), fueron los más frecuentes. La proporción del área oscura basal (DS-III2 ) y del tarsómero posterior (Ta-III2 ) constituyó una característica sólida de diagnóstico, pues representó el 89 % (n=213/240) del total de especímenes analizados. Se encontraron diferencias significativas en la forma (F=1,65; gl=50; p<0,001) y el tamaño (F=3,37; gl=5; p=0,005) del ala entre las poblaciones de diferentes localidades. El centroide de menor tamaño (2,64 mm) se encontró en las poblaciones de Chocó. Conclusiones. Se registraron 11 patrones nuevos para las manchas de la vena costal y se confirmó la dominancia de los patrones alares I y VI en las poblaciones de An. darlingi de Colombia, así como la relación de DS-III2 y Ta-III2 como un rasgo sólido de diagnóstico para la taxonomía de la especie. Se encontraron diferencias en el tamaño corporal de las poblaciones evaluadas, lo cual reviste importancia para el análisis de aspectos bionómicos de la especie.

Abstract Introduction: Natural populations of Anopheles darlingi, the main malaria vector in Colombia, have shown phenotypic variations in some of their diagnostic characters. Objective: To characterize morphometric variations in wing spot patterns and diagnostic characters of the hind leg of adult females of An. darlingi collected in areas where malaria is endemic. Materials and methods: Patterns of the costal vein spots of wild females of An. darling populations collected in the departments of Chocó, Guaviare, Meta and Vichada were analyzed using linear and morphometric approaches. The second tarsomere of the hind leg of females was characterized by linear morphometric analysis. Results: We found 19 patterns of spots in the costal vein; patterns I (n=118/240, 49%) and VI (n=66, 28%) were the most frequent. The proportion of the basal dark area of hind tarsomere II and the length of hind tarsomere II (DSIII2 /Ta-III2) constituted a robust diagnostic character as it represented 89% (n=213/240) of the total specimens studied. Significant differences were found in the wing shape (F=1.65, df =50, p<0.001) and the wing size (F=3.37, df=5, p=0.005) among populations from different locations. The smallest centroid size (2.64 mm) was found in populations from Chocó. Conclusions. We registered 11 new wing spot patterns in the costal vein and the dominance of the patterns I and VI for populations of An. darlingi from Colombia. We confirmed DSIII2/TaIII2 ratio as a robust diagnostic character for the taxonomy of this species. We found differences between the size and shape of the wings of An. darlingi populations in accordance to their geographical distribution, which constitute important bionomic aspects for this malaria vector.

Análise da estrutura populacional de mosquitos Aedes aegypti (Diptera: Culicidae) em diferentes estratos urbanos na cidade de São Paulo, utilizando morfometria geométrica alar e marcadores microssatélites / Analysis of the population structure of Aedes aegypti (Diptera: Culicidae) mosquitoes in different urban strata in the city of São Paulo, using wing geometric morphometry and microsatellite markers

O mosquito Aedes aegypti é reconhecido como o principal vetor do vírus da Dengue, além de transmitir outros arbovírus de importância médica, como os causadores da Febre Amarela urbana, Chikungunya e Zika. A ecologia deste vetor está intimamente associada ao homem, sendo provavelmente, a única espécie de culicídeo a conseguir completar todo seu ciclo de vida dentro das habitações humanas, com sua dinâmica populacional fortemente relacionada aos processos decorrente da urbanização. Assim como outras metrópoles, a cidade de São Paulo apresenta estresse ambiental, em função da elevada densidade populacional e urbanização não planejada, o que contribui para a proliferação do Ae. aegypti e, consequentemente, o aumento no número de casos de Dengue. Embora, vacinas como a da Febre Amarela e Dengue já tenham sido desenvolvidas, esta última mais recentemente e, ainda não empregada em larga escala, o controle do vetor ainda permanece como a principal estratégia para a disruptura dos padrões epidemiológicos das arboviroses causadas por seus patógenos. Estruturação populacional, geralmente é um resultado de combinações decorrentes de processos históricos e contemporâneos envolvendo determinada espécie, como a sua capacidade de dispersão, padrões de cópula, barreiras físicas e ambientais, além de padrões demográficos. Desse modo, determinar os diferentes papéis destes processos na estruturação de populações torna-se útil no controle de vetores de importância médica. Um bom exemplo é a propagação da resistência a inseticidas, em decorrência do fluxo gênico entre as populações. Portanto, um melhor conhecimento da estruturação populacional do Ae. aegypti é crucial para auxílio e desenvolvimento de novas estratégias de controle. Dessa forma, visando elucidar seu padrão de estruturação, o presente estudo utilizou-se da morfometria geométrica alar e de marcadores microssatélites, para investigação de 11 populações de mosquitos Ae. aegypti coletados em áreas com diferentes graus de urbanização, localizadas no município de São Paulo. Os resultados encontrados sugerem um padrão de estruturação de acordo com o gradiente de urbanização no qual os espécimes foram coletados. As distâncias de Mahalanobis, obtidas pela morfometria geométrica alar, apresentaram significância estatística em 54 dos 55 testes conduzidos, com as populações exibindo um claro padrão de segregação nas Análises de Variáveis Canônicas e Neighbor-Joining, tanto para as populações agrupadas na forma de seus estratos urbanos, como por seus respectivos locais de coleta, enquanto que o teste de reclassificação dos espécimes alcançou relativo grau de precisão de reconhecimento. Os microssatélites indicaram uma baixa estruturação genética (Fst = 0,057), com 93 por cento de seus valores apresentando significância estatística. Contudo, em conformidade com o gradiente de urbanização dos estratos, com moderado fluxo gênico, déficit de heterozigosidade e indícios de expansão populacional, principalmente nas áreas com maior grau de urbanização. A intensificação dos processos decorrentes da urbanização tem como causa a diminuição dos espaços verdes encontrados nas cidades, de modo a contribuir para a elevação da temperatura e, consequentemente, favorecer a proliferação do Ae. aegypti. Adicionalmente, a perda destes espaços implica no processo de homogeneização biótica, fenômeno que atua como adjuvante a plasticidade ecológica do vetor, de maneira a beneficia-lo. Hipótese, corroborada pela sua expansão populacional, exibida principalmente nos ambientes mais antropizados. A estruturação observada nas populações de Ae. aegypti no presente estudo indica que os processos de urbanização desempenham um importante papel na sua conformação, e fatores como o moderado fluxo gênico e déficit de heterozigosidade podem estar refletindo nos seus padrões epidemiológicos

Aedes aegypti is recognized as the main vector of Dengue, in addition to transmit other arboviruses of medical importance, as the agents of Yellow Fever, Chikungunya and Zika. The ecology of this vector is closely associated with the human, being probably the only kind of mosquito to be able to complete all their life cycle inside the human habitations, with their population dynamics strongly related to processes arising from urbanization. Like other cities, the city of São Paulo suffers from environmental stress due to the high population density and unplanned urbanization, which contributes to the proliferation of Ae. aegypti mosquitoes and consequently the increase in the number of cases of dengue fever. Although vaccines such as Yellow Fever and Dengue have already been developed, the latter, more recently, and not yet used on a large scale, vector control remains the main strategy for the disruption of epidemiological patterns of arboviral diseases caused by their pathogens. Structure of the population, is generally the result of combinations resulting from historical and contemporary processes involving certain species, such as their ability to disperse, copulation pattern, physical and environmental barriers and demographic trends, and to determine the different roles of these processes in structuring the population becomes very useful for the medical importance of vector control. A good example is the spread of insecticide resistance, due to gene flow between populations. Therefore, a better understanding of the population structure of Ae. Aegypti is crucial to support and develop new strategies for control programs. Thus, in order to elucidate its pattern of structuring this study utilized wing geometric morphometric and microsatellite markers, for investigation of 11 Ae. aegypti populations collected in areas with different degrees of urbanization, located in the municipality of São Paulo. The results suggest a pattern of structuring according to the urbanization gradient in which the specimens were collected. The distances of Mahalanobis obtained by wing geometric morphometry, statistically significant in 54 of the 55 tests performed, with populations showing a clear trend of segregation in the Canonical Variables analysis and Neighbor-Joining, both for the populations grouped in the form of their urban strata as per their respective collection locations, while the reclassification of test specimens reached relative degree of recognition accuracy. Microsatellites indicated a low genetic structure (Fst = 0.057), with 93 per cent of their statistically significant values. However, in accordance with the gradient of urbanization of the strata, with moderate gene flow, heterozygosity and evidence of population expansion, especially in the areas with the highest degree of urbanization. The intensification of the processes resulting from urbanization Implies in the reduction of the green spaces found in the cities, in order to contribute to the increase of the temperature and thus the proliferation of the Ae. Aegypti. In addition, the loss of these spaces involves biotic homogenization process, a phenomenon that acts as an adjuvant ecological plasticity of the vector, in order to benefit it. Hypothesis, corroborated by its population expansion, displayed mainly in anthropic environments. The structure observed in populations of Ae. aegypti in this study indicates that the urbanization processes play an important role in their conformation, and factors such as moderate gene flow and deficit of heterozygosity can be reflected in their epidemiological patterns

Análise da estrutura populacional de mosquitos Culex quinquefasciatus e Culex nigripalpus (Diptera: Culicidae) utilizando marcadores de microssatélites e análise de morfometria geométrica alar / Analysis of the population structure of Culex quinquefasciatus and Culex nigripalpus (Diptera: Culicidae) mosquitoes using microsatelllite markers and morphometric geometric analysis of the wing

Introdução: Parques inseridos na malha urbana de grandes metrópoles possuem potencial para manter o ciclo biológico de diversas espécies vetoras de patógenos, como as espécies Culex quinquefasciatus e Culex nigripalpus. Consideradas antropofílicas, essas espécies têm importância epidemiológica e são abundantemente encontradas na cidade de São Paulo. Porém, pouco se sabe sobre as características genéticas dessas espécies em escala microgeográfica. Visando o melhor entendimento sobre a estrutura populacional dessas espécies, foi analisado os padrões da forma alar e a caracterização genética por marcadores de microssatélites, afim de se obter informações que contribuam para o entendimento da situação populacional desses vetores dentro do município. Objetivos: (1) Analisar a variabilidade da forma alar nas populações de Cx. quinquefasciatus e Cx. nigripalpus; (2) Analisar a variabilidade genética e o fluxo gênico nas mesmas populações; (3) Testar a funcionabilidade de primers desenvolvidos para regiões de microssatélites nas populações de Cx. nigripalpus. Material e Métodos: No total, foram estudadas cinco populações de Cx. quinquefasciatus e sete populações de Cx. nigripalpus, coletados em parques urbanos da cidade de São Paulo. Análises discriminantes, como variável canônica, teste de reclassificação cruzada e dendrograma de Neighbor-joining, utilizando os software Morpho J e Past, foram realizadas para a compreensão do formato e tamanho da asa direita nas populações. Em relação ao estudo da estruturação genética, foram testados 12 pares de primers de microssatélites em mosquitos Cx. quinquefasciatus e 33 pares de primers em mosquitos Cx. nigripalpus. Resultados: Análise da morfometria geométrica alar nas populações de Cx. quinquefasciatus demonstrou homogeneidade nos formatos alares, sendo um caracter preservado nessa espécie pela cidade, contudo, há tênues diferenças na população coletada em ambiente mais silvestre. O mesmo foi observado para as populações de Cx. nigripalpus, onde foi possível visualizar uma subestruturação na forma alar dentro da população Shangrilá. Dos primers testados, 12 amplificaram de forma consistente em todas as populações de Cx. quinquefasciatus e seis primers nas populações de Cx. nigripalpus. Os resultados encontrados sugerem que ambas as espécies possuem baixa estruturação genética, com fluxo gênico moderado entre as populações de Cx. quinquefasciatus e baixo entre as de Cx. nigripalpus, apresentando alto índice de heterozigosidade, onde as únicas populações que estão em expansão são as que foram coletadas em ambientes onde a urbanização está avançando. Discussão: Processos de urbanização, somados às mudanças causadas no ambiente, beneficiam e tendem a elevar a abundância dessas espécies em ambientes antropizados. A baixa estruturação genética e morfométrica das asas dessas espécies indicam a adaptação delas na cidade, havendo segregação devido à heterogeneidade do ambiente em que as populações se encontram. Conclusão: Evidências de baixa estruturação entre as populações e indícios de expansão em populações de ambientes mais silvestres indicam que essas espécies estão atreladas ao processo de urbanização da cidade de São Paulo

Introduction: Urban parks have the potential to harbor and maintain the life cycle of several mosquitoes such as Culex quinquefasciatus and Culex nigripalpus, both are anthropophilic species with epidemiological importance and very abundant in São Paulo city. However, their genetics characteristics are poorly know. Aiming the better understanding of the population structure of these species in microregion, was evaluated the wing shape variation and selected microsatellite loci. In this way, the information obtained can contribute to the understanding of the population situation of these vectors in São Paulo city. Objectives: (1) To evaluate the wing shape variability in populations of Cx. quinquefasciatus and Cx.nigripalpus; (2) To evaluate the genetic variability and gene flow in the Cx. quinquefasciatus and Cx.nigripalpus populations (3) Test microsatellite markers functionality parameters in Cx. nigripalpus, previously used successfully in other Culex species. Material and Methods: Were studied, five populations of Cx. quinquefasciatus and seven population of Cx. nigripalpus collected in urban parks in São Paulo city. Discriminant analysis was made to evaluate the wing shape patterns, such as Canonical variate analysis, cross validated test and Neghbor-joninin dendrogram using Morpho J and Past softwares, were perfomed to understand the size and shape of the right wing in these populations. For the study of genetic structuring, there were tested 12 pairs of microsatellite loci in Cx. quinquefasciatus samples and 33 pairs of microsatellite loci in Cx. nigripalpus samples. Results: The wing shape patterns in Cx. quinquefasciatus population were homogeneous, showing a preserved character in this population, however, has been tenuous differences in the more sylvatic population. The same pattern was observed in Cx. nigripalpus populations and was observed substructuring in the Shangrilá population. From the tested primers, 12 were functional and amplified consistently in the all five Cx. quinquefasciatus population and for Cx. nigripalpus, six primers were amplified. The results suggest both species having low genetic structure, moderate gene flow and the only populations which are expading were collected in areas that the urbanization is increasing. Discussion: Urbanization processes added to the environmental changes benefit and tend to raise the abundance of these species in anthropized locals. The low genetic structure and alar morphometry indicates the adaptation of these species in São Paulo city, there being segregated due to the environment heterogeneity which the population inhabit. Conclusion: Evidence of low structure between populations studied and signs of expansion in populations of more sylvatic environments indicate that these species are linked to urbanization process

Morphometric differences and fluctuating asymmetry in Melipona subnitida Ducke 1910 (Hymenoptera: Apidae) in different types of housing / Diferenças morfométricas e assimetria flutuante em Melipona subnitida Ducke 1910 (Hymenoptera: Apidae) em diferentes tipos de habitação

Abstract A geometric morphometrics approach was applied to evaluate differences in forewing patterns of the Jandaira bee (Melipona subnitida Ducke). For this, we studied the presence of fluctuating asymmetry (FA) in forewing shape and size of colonies kept in either rational hive boxes or natural tree trunks. We detected significant FA for wing size as well as wing shape independent of the type of housing (rational box or tree trunks), indicating the overall presence of stress during the development of the studied specimens. FA was also significant (p < 0.01) between rational boxes, possibly related to the use of various models of rational boxes used for keeping stingless bees. In addition, a Principal Component Analysis indicated morphometric variation between bee colonies kept in either rational hive boxes or in tree trunks, that may be related to the different origins of the bees: tree trunk colonies were relocated natural colonies while rational box colonies originated from multiplying other colonies. We conclude that adequate measures should be taken to reduce the amount of stress during bee handling by using standard models of rational boxes that cause the least disruption.

Resumo A abordagem da morfometria geométrica foi aplicada para avaliar as diferenças nos padrões das asas anteriores da abelha Jandaíra (Melipona subnitida Ducke). Para isso, estudou-se a presença de assimetria flutuante (AF) na forma das asas anteriores e tamanho das colônias mantidas tanto em caixas de colméia racional ou troncos de árvores naturais. Foi detectado AF significativa para o tamanho da asa, bem como a forma da asa independente do tipo de habitação (caixa racional ou cortiço), indicando a presença global de estresse durante o desenvolvimento dos espécimes estudados. AF também foi significativa (p < 0,01) entre as caixas racionais, possivelmente relacionados com a utilização de vários modelos de caixas racionais utilizados para a conservação de abelhas sem ferrão. Além disso, a Análise de Componentes Principais indicou variações morfométricas entre as colônias de abelhas mantidos em caixas racionais ou em cortiços, que podem estar relacionados com as diferentes origens das abelhas: os cortiços foram colônias naturais realocados, enquanto as colônias das caixas racionais foram originadas da multiplicação outras colônias. Conclui-se que devem ser tomadas medidas adequadas para reduzir a quantidade de estresse durante o manuseio abelha usando modelos padrão de caixas racionais que causam a menor perturbação.

Can body traits, other than wings, reflect the flight ability of Triatominae bugs?

Abstract: INTRODUCTION : Insects of the subfamily Triatominae are vectors of Trypanosoma cruzi , the Chagas disease parasite, and their flying behavior has epidemiological importance. The flying capacity is strikingly different across and within Triatominae species, as well as between sexes or individuals. Many Triatoma infestans individuals have wings but no flying muscles. In other Triatominae species, no clear relationships were found between wing length and flying behavior. If wing presence or size is not reflective of the flying behavior, which other parts of the body could be considered as reliable markers of this important function? METHODS : The genus Mepraia has exceptional characteristics with invariably wingless females and wingless or winged males. We calculated the porous surface exposed to odorant molecules to estimate the olfactory capacity of Mepraia spinolai . The head shape and thorax size were estimated using the geometric morphometric approach and traditional morphometric techniques, respectively. RESULTS : Alary polymorphism in M. spinolai was significantly associated with consistent modification of the thorax size, head shape, and notable change in the estimated olfactory capacity. The macropterous individuals had a larger olfactory surface and thorax size and significantly different head shape compared to those of the micropterous individuals. CONCLUSIONS: We concluded that these structural changes could be associated with the flying potential of Triatominae. Thus, morphological attributes not found on wings could help determine the likely flying potential of the bugs.

Fluctuating asymmetry in Apis mellifera(Hymenoptera: Apidae) as bioindicator of anthropogenic environments / Asimetría fluctuante en Apis mellifera (Hymenop-tera: Apidae) como bioindicador de ambientes antropo-génicos

The successful distribution of A. melliferais due to their ability to adjust to seasonal variations, considerable control over their internal physical environment and exploration of different resources. However, their populations have experienced different forms and levels of environmental pressure. This research aimed to verify the phenotypic plasticity in both size and shape of wings in A. melliferausing fluctuating asymmetry, based on geometric morphometrics from apiaries located in sites with high and low levels of anthropization. We sampled 16 locations throughout all five geographic regions of Brazil. At each site, samples were collected from 20 beehives installed in apiaries: 10 installed near high anthropogenic environments (Cassilàndia - MS, Fortaleza - CE, Maringá - PR, Aquidauana - MS, Rolim de Moura - RO, Riachuelo - SE, Ubirata - PR and Piracicaba - SP), and 10 in sites with low levels of human disturbance (Cassilàndia - MS, Itapiúna CE, Uniao da Vitoria - PR, Aquidauana - MS, Rolim de Moura - RO, Pacatuba - SE, Erval Seco - RS, Rio Claro - SP). A sample of 10 individuals was taken in each hive, totaling 200 per location, for a total of 1 600 individuals. We used fluctuating asymmetry (FA) in size and shape of the forewing through geometric morphometrics. The FA analysis was conducted in order to check bilateral differences. The indexes of size and shape were submitted to analysis of variance (ANOVA), where the characters evaluated were used as factors to verify the size and shape differences. The results indicated an asymmetry on the shape of the wing (P < 0.001) but no asymmetry was observed on wing size. Considering FA as an environmental response and high and low impacted areas as a fixed factor, we observed significant differences (P < 0.05). The results for the wing shape in A. melliferademonstrated that this feature undergoes more variation during ontogeny compared to the variation in size. We concluded that bee samples collected from colonies with higher levels of human disturbance had higher wing-shape asymmetry; the variation of fluctuating asymmetry in the wing shape of honeybees can be used as an indicator of the degree of environmental anthropization.

La distribución exitosa de A. melliferase debe a su capacidad para adaptarse a las variaciones estacionales, controlar considerablemente su ambiente físico interno y por la exploración de recursos. Sin embargo, sus poblaciones experimentan diferentes formas y niveles de presión ambiental. Esta investigación evaluó colmenares, ubicadas en entornos con distintos niveles de antropización. Ambientes considerados altamente antropogénicos fueron escogidos: áreas urbanas, agrícolas con aplicaciones de insecticidas, y las industrias con grandes áreas (regiones con altos índices de contaminación). Por otra parte, los entornos considerados con bajos niveles de perturbación humana fueron elegidos: áreas de preservación permanente de los bosques restantes (Mata Atlántica, Planalto, Cerrado), áreas ribereñas o reservas ecológicas. Se muestrearon 16 localidades que abarcan las cinco regiones del Brasil. Estamos utilizando la asimetría fluctuante desde el tamaño y la forma del ala anterior por técnicas de morfometría geométrica. En cada sitio, las muestras se obtuvieron de las 20 colmenas instaladas en los colmenares: 10 instalados cerca de entornos altamente antropogénicos (Cassilándia - MS, Fortaleza - CE, Maringá - PR, Aquidauana - MS, Rolim de Moura - RO, Riachuelo - SE, Ubirata - PR y Piracicaba - SP) y 10 en sitios de bajo nivel de perturbación humana (Cassilándia - MS, ItapiúnaCE, Uniao da Vitória - PR, Aquidauana - MS, Rolim de Moura - RO, Pacatuba- SE, Erval Seco - RS, Rio Claro - SP). Los índices de tamaño y forma fueron sometidos a análisis de varianza (ANOVA), donde se utilizaron los caracteres evaluados como factores, para verificar las diferencias de tamaño y forma. La asimetría de análisis fluctuante (AF) se llevó a cabo con el fin de comprobar las diferencias bilaterales. Los resultados indican la existencia de la asimetría de la forma del ala (P < 0.001), pero no se observó asimetría del tamaño del ala. Considerando AF como respuesta ambiental y áreas de alto y bajo grado de alteración humana como factor fijo, observamos diferencias significativas (P < 0.05). Los resultados, para la forma de ala de la A. mellifera,muestran que esta característica se somete a más variación durante la ontogenia en comparación con la variación en el tamaño. Llegamos a la conclusión de que las colonias de abejas recogidas en ambientes con niveles más altos de la perturbación humana tienen una mayor asimetría en forma de ala, por lo que la asimetría fluctuante en forma de alas de las abejas puede ser utilizada como un indicador del grado de antropización del medio ambiente.

Caracterização de populações de Culex coronator (Diptera: Culicidae) e distinção de fêmeas adultas de Culex coronator e Culex usquatus por meio da análise da morfometria geométrica de asa e de sequências gênicas / Characterization of populations of Culex coronator (Diptera: Culicidae) and differentiation of adult females of Culex coronator and Culex usquatus using wing geometric morphometry and gene sequences

Culex coronator Dyar & Knab e Culex usquatus Dyar são duas espécies irmãs, que fazem parte do Complexo Coronator, composto por mais quatro espécies (Culex usquatissimos Dyar, Culex ousqua Dyar, Culex camposi Dyar, Culex covagarciai Forattini). As fêmeas deste grupo são indistinguíveis por caracteres morfológicos, sendo a identificação possível somente através da distribuição e tamanho das cerdas apicais que ornamentam o gonocoxito da genitália masculina. Cx. coronator, é a espécie com maior distribuição geográfica, ocupando as Américas do Norte, Central e Sul. Já Cx. usquatus só foi registrado nas Américas Central e do Sul, ocorrendo em simpatria no Brasil com Cx. coronator. Apesar da semelhança morfológica das fêmeas das duas espécies, até o momento, somente Cx. coronator foi encontrado naturalmente infectado por diversas arboviroses. Considerando que estudos populacionais são importantes para compreender a evolução e a dinâmica de populações de pontencias vetores, e que a correta identificação de fêmeas é fundamental para estudos de competência vetorial, os objetivos deste trabalho foram: (1) distinguir fêmeas adultas de Cx. coronator de Cx. usquatus (2) obter conhecimento da estrutura populacional de Culex coronator nas regiões Sul e Sudeste (3) examinar a possível existência de outras espécies não descritas e/ou incorretamente identificadas sob o epíteto de Cx. coronator. Para tanto foram utilizadas duas ferramentas: uma morfológica (morfometria de asa), e outra genética...

Culex coronator and Culex usquatus are sibling species belonging to Coronator Group, which comprises five other species (Culex usquatissimos Dyar, Culex ousqua Dyar, Culex camposi Dyar, Culex covagarciai Forattini). Except by Cx. yojae, the females of this group are indistinguishable, being the identification only possible by the analysis of the arrangement and number of appendicles on the apical lobe of the gonocoxite of the male genitalia. Cx. coronator is the most widely distributed species in the complex, occupying North, Central and South America, while Cx. usquatus was recorded only in Central and South America. Therefore, these species are sympatric in Brazil. Despite the morphological similarity of the females of both species, only Cx. coronator has epidemiological importance, being found infected with many arboviruses. Since studies focusing in population structure are important to understand the evolution and dynamics of potencial vectors and that the correct female identification is critical for development of vectorial competence studies, the aims of this study were to: (1) distinguish adult females of Cx. coronator from Cx. usquatus (2) obtain knowledge of the population structure of Culex coronator in Southern and Southeastern areas (3) examine the presence of undescribed and/or incorrectly identified species under Cx. coronator. Thereby, a survey was carried out using morphological (wing geometry) and genetic (4 microsatellite loci and barcode region...

Temporal variation of wing geometry in Aedes albopictus

Although native to the tropical and subtropical areas of Southeast Asia, Aedes albopictus is now found on five continents, primarily due to its great capacity to adapt to different environments. This species is considered a secondary vector of dengue virus in several countries. Wing geometric morphometrics is widely used to furnish morphological markers for the characterisation and identification of species of medical importance and for the assessment of population dynamics. In this work, we investigated the metric differentiation of the wings of Ae. albopictus samples collected over a four-year period (2007-2010) in São Paulo, Brazil. Wing size significantly decreased during this period for both sexes and the wing shape also changed over time, with the wing shapes of males showing greater differences after 2008 and those of females differing more after 2009. Given that the wings play sex-specific roles, these findings suggest that the males and females could be affected by differential evolutionary pressures. Consistent with this hypothesis, a sexually dimorphic pattern was detected and quantified: the females were larger than the males (with respect to the mean) and had a distinct wing shape, regardless of allometric effects. In conclusion, wing alterations, particularly those involving shape, are a sensitive indicator of microevolutionary processes in this species.

Morfometría geométrica alar para la identificación de Chrysomya albiceps y C. megacephala (Diptera: Calliphoridae) de Venezuela / Geometric wing morphometrics for Chrysomya albiceps and C. megacephala identification (Diptera: Calliphoridae) from Venezuela

Calliphoridae is one of the families with the greatest number of species with forensic importance, which immature stages feed and develop on decaying material. in Venezuela, there are few studies on this taxon reporting the dominance of Chrysomya over other carrion flies. The goal of this study was to analyze the variations on wing morphometrics, to support the identification of two forensic flies. For this we photographed a total of 168 wings from C. albiceps (n=111) and C. megacephala (n=57). Landmark coordinate (x, y) configurations were registered and aligned by Generalized Procrustes Analysis. Principal Component Analysis and shape significance test based on Procrustes distance were implemented. Statistical analysis of variance found significant differences in wing isometric size (Kruskal-Wallis). The PCA showed the separation between species, and shape test showed significant differences (F Goodall´s). The main differences between both species were localized on subcosta rupture, join of R2+3 with wing border, join of dm-cu, and join of Cu with bm-cu, suggesting that wing morphometrics is a suitable tool in the discrimination of blowfly species.

Calliphoridae es una de las familias con el mayor número de especies de importancia forense, donde sus fases inmaduras se alimentan y desarrollan sobre material en descomposición. En Venezuela, son pocos los estudios en este taxon, sin embargo se ha reportado la dominancia del género Chrysomya sobre los otros dípteros. El objetivo fue analizar la variación morfométrica en la arquitectura alar, como herramienta de apoyo a la identificación de dípteros de importancia forense. Se fotografiaron 168 alas de C. albiceps (n=111) y C. megacephala (n=57) y se registraron configuraciones de coordenadas (x, y), se alinearon mediante Análisis Generalizado de Procrustes. Se efectuaron Análisis de Componentes Principales y comparaciones pareadas entre distancia parcial de Procrustes. El análisis estadístico de varianza encontró diferencias en el tamaño isométrico del ala (Kruskal-Wallis). El ACP mostró la separación de ambas especies, y la prueba de configuraciones determinó diferencias significativas (F Goodall). Las principales diferencias entre ambas especies se encontraron en: ruptura de la subcosta, unión de R2+3 con el borde del ala, unión dm-cu, y unión de Cu con dm-cu, lo que confirma que la morfometría alar es una herramienta idónea en la discriminación de especies de Calliphoridae.

Caracterização de populações de Anopheles darlingi (Diptera/Culicidae) do Brasil por estruturas de morfologia externa dos ovos, das asas e por sequências gênicas / Characterization of Anopheles darlingi (Diptera: Culicidae) populations from Brazil by external morphological structure of eggs, wings and gene sequences

Introdução - A malária é uma das principais doenças humanas do mundo e afeta principalmente as populações pobres em regiões tropicais e subtropicais onde as condições ambientais são favoráveis tanto para a proliferação dos agentes etiológicos como dos mosquitos vetores. No Brasil, An. darlingi é considerado vetor primário de plasmódios humanos. Devido à importância médica, esse inseto tem sido objeto de campanhas de controle populacional. No entanto, não foi considerada a ocorrência de microevolução em An. darlingi e consequentemente, a possibilidade de populações diferentes apresentarem características biológicas distintas. Objetivo Verificar a presença de variabilidade genética e morfológica em populações de An. darlingi no Brasil. Metodologia - Foram analisados e comparados 26 atributos da morfologia externa de ovos de nove populações, bem como a morfometria geométrica da forma alar de espécimes de 10 localidades diferentes de Anopheles darlingi. Além disso, foram empregadas sequências do gene mitocondrial citocromo oxidase subunidade I (COI) para analisar a estrutura populacional de An. darlingi. Resultados e Conclusão Apesar dos atributos dos ovos apresentarem variação, somente as amostras de Tocantins e Pará foram diferenciadas das demais populações. As variações nas estruturas externas dos ovos são provavelmente adaptativas, com influência de fatores ambientais, como temperatura, umidade e disponibilidade de alimento para as fêmeas. A comparação da morfometria geométrica da asa demonstrou que existe maior similaridade entre as populações da costa (estados do Espírito Santo , do cerrado (estados de Goiás e Tocantins), do norte do rio Amazonas (estados do Amazonas e Amapá), e do sul do rio Amazonas (estados de Mato Grosso e Pará).

Molecules, wing pattern and distribution: an approach to species delimitation in the "loxurina group" (Lepidoptera: Lycaenidae: Penaincisalia)

The wide range of morphological variations in the "loxurina group" makes taxa identification difficult, and despite several reviews, serious taxonomical confusion remains. We make use of DNA data in conjunction with morphological appearance and available information on species distribution to delimit the boundaries of the "loxurina" group species previously established based on morphology. A fragment of 635 base pairs within the mtDNA gene cytochrome oxidase I (COI) was analysed for seven species of the "loxurina group". Phylogenetic relationships among the included taxa were inferred using maximum parsimony and maximum likelihood methods. Penaincisalia sigsiga (Bálint et al), P. cillutincarae (Draudt), P. atymna (Hewitson) and P. loxurina (C. Felder & R. Felder) were easily delimited as the morphological, geographic and molecular data were congruent. Penaincisalia ludovica (Bálint & Wojtusiak) and P. loxurina astillero (Johnson) represent the same entity and constitute a sub-species of P. loxurina. However, incongruence among morphological, genetic, and geographic data is shown in P. chachapoya (Bálint & Wojtusiak) and P. tegulina (Bálint et al). Our results highlight that an integrative approach is needed to clarify the taxonomy of these neotropical taxa, but more genetic and geographical studies are still required.

Taxonomic considerations on the genera Moneuptychia Forster and Carminda Dias, reval. (Lepidoptera, Nymphalidae, Satyrinae) / Considerações taxonômicas sobre os gêneros Moneuptychia Forster e Carminda Dias, reval. (Lepidoptera, Nymphalidae, Satyrinae)

Euptychia soter Butler, 1877, the type species of Moneuptychia Forster, 1964, is compared to Satyrus paeon Godart, 1824, the type species of Carminda Dias, 1998. The male genitalia and wing design patterns of these species are dealt with. Some additional data from other species from both genera are also presented. The morphological comparisons carried out in this study indicate that Carminda is not a junior synonym of Moneuptychia. Thus, Carminda is revalidated.

Euptychia soter Butler, 1877, espécie-tipo de Moneuptychia Forster, 1964, é comparada com Satyrus paeon Godart, 1824, espécie-tipo de Carminda Dias, 1998. A genitália do macho e o padrão de desenho das asas dessas espécies são comparados. Alguns dados sobre outras espécies de ambos os gêneros são também incluídos. As comparações morfológicas ora apresentadas mostram que Carminda não é um sinônimo júnior de Moneuptychia. Portanto, Carminda é revalidado.

Resource Defense Polygyny by Hetaerina rosea Selys (Odonata: Calopterygidae): Influence of Age and Wing Pigmentation

Current evidence suggests that in Hetaerina damselflies males exhibit lek mating system. In this study, in order to answer if the same occurs in Hetaerina rosea Selys, we manipulated vegetation substrates used as territories and quantified the number of visiting females, males defending territories and fight intensity. We also examined whether body size and wing pigmentation are selectable traits in male-male competition, and if age affects male territorial behavior. Our results showed that males with larger pigmented areas won more contests, independently of body size. Old males changed from territoriality to sneaking strategy. Contrary to other Hetaerina species, males of H. rosea do not display lek behavior, but defend resources according to the resource defense polygyny strategy.

The balance between GMD and OFUT1 regulates Notch signaling pathway activity by modulating Notch stability

The Notch signaling pathway plays an important role in development and physiology. In Drosophila, Notch is activated by its Delta or Serrate ligands, depending in part on the sugar modifications present in its extracellular domain. O-fucosyltransferase-1 (OFUT1) performs the first glycosylation step in this process, O-fucosylating various EGF repeats at the Notch extracellular domain. Besides its O-fucosyltransferase activity, OFUT1 also behaves as a chaperone during Notch synthesis and is able to down regulate Notch by enhancing its endocytosis and degradation. We have reevaluated the roles that O-fucosylation and the synthesis of GDP-fucose play in the regulation of Notch protein stability. Using mutants and the UAS/Gal4 system, we modified in developing tissues the amount of GDP-mannose-deshydratase (GMD), the first enzyme in the synthesis of GDP-fucose. Our results show that GMD activity, and likely the levels of GDP-fucose and O-fucosylation, are essential to stabilize the Notch protein. Notch degradation observed under low GMD expression is absolutely dependent on OFUT1 and this is also observed in Notch Abruptex mutants, which have mutations in some potential O-fucosylated EGF domains. We propose that the GDP-fucose/OFUT1 balance determines the ability of OFUT1 to endocytose and degrade Notch in a manner that is independent of the residues affected by Abruptex mutations in Notch EGF domains.

Morphological analysis of three populations of Anopheles (Nyssorhynchus) nuneztovari Gabaldón (Diptera: Culicidae) from Colombia

Based on the results of comparative analyses of 1,039 specimens of several progenies of Anopheles nuneztovarifrom three localities in Colombia, eight costal wing spot patterns were observed. Patterns I and III were the most frequent: 77.96 percent and 11.36 percent, respectively. Using the diagnostic characters ratio of the length of the basal dark area of hind tarsomere II/length of hind tarsomere II, ratio of the length of the humeral pale spot/length of the pre-humeral dark spot, and the ratio of the length of the subcostal pale spot/length of the distal sector dark spot (DS-III2/Ta-III2, HP/PHD, SCP/DSD) approximately 5 percent of the adult females were misidentified as a species of Nyssorhynchus, different from An. nuneztovari. Approximately 5 percent of the specimens showed DS-III2/Ta-III2 ratio less than 0.25 (range 0.21 - 0.24), and among them 3.34 percent shared a HP/PHD ratio less than 1.50. Consequently, 1.52 percent of An. nuneztovari individuals can be misidentified as Anopheles oswaldoi. In those specimens with the DS-III2/Ta-III2 ratios higher than 0.25, 34.45 percent displayed SCP/DSD values greater than 0.50 and of these, 3.65 percent displayed HP/PHD values greater than 1.8. This combination of characters could lead one to misidentify samples of An. nuneztovari as Anopheles rangeli. Similarly, 2.43 percent of the females could be identified erroneously as either Anopheles aquasalis or Anopheles benarrochi. Individuals with a HP/PHD ratio greater than 2.0, could be misidentified as Anopheles trinkae, Anopheles strodei or Anopheles evansae. A distinct combination of diagnostic characters for An. nuneztovari from Colombia is proposed.

Curvature measurement as a tool for the analysis of morphometric variation using Drosophila wings as a model

This article describes the use of geometric measurements of continuous, discrete parameters to studymorphometric variation in the wing cells of two sibling species, Drosophila mercatorum and Drosophilaparanaensis. To validate the results, the same wing samples were also analyzed using truss networks andpartial warps, in addition to a comparison with the ellipse method. The use of discriminative measurementsin conjunction with a Bayesian-based classification method yielded a relatively high number of correctclassifications for new individuals. These results compared favorably with those obtained using trussnetworks, partial warps and the ellipse method. These findings indicate that continuous curvature and arclength measurements may be useful parameters for the morphometric analysis of insect wings and possiblyother biological structures and shapes.

Morphometric analysis of Triatoma dimidiata populations (Reduviidae: Triatominae) from Mexico and Northern Guatemala

Triatoma dimidiata is one of the major vectors of Chagas disease in Latin America. Its range includes Mexico, all countries of Central America, Colombia, and Ecuador. In light of recent genetic analysis suggesting that the possible origin of this species is the Yucatan peninsula, we have analyzed populations from the state of Yucatan, San Luis Potosi, and Veracruz in Mexico, and a population from the southern region of the Yucatan peninsula located in Northern Guatemala, the region of El Peten. Classical morphometry including principal component, discriminant, sexual dimorphism, and wing asymmetry was analyzed. San Luis Potosi and Veracruz populations were indistinguishable while clearly separate from Yucatan and Peten populations. Despite important genetic differences, Yucatan and Peten populations were highly similar. Yucatan specimens were the smallest in size, while females were larger than males in all populations. Only head characters were necessary to distinguish population level differences, although wing fluctuating asymmetry was present in all populations. These results are discussed in light of recent findings suggesting genetic polymorphism in most populations of Triatoma dimidiata south of Chiapas to Ecuador.

Morfometria de Papilioninae (Lepidoptera, Papilionidae) ocorrentes em quatro localidades do Rio Grande do Sul, Brasil. I. Comparações com a massa corporal / Morphometrics of Papilioninae (Lepidoptera, Papilionidae) occurring in four localities of Rio Grande do Sul, Brazil. I. Comparisons with body mass

A análise da relação entre a asa e partes do corpo em Lepidoptera podem revelar uma conexão entre a estrutura e a função. Este estudo investigou relações entre o tamanho do corpo de papilionídeos e suas partes, importantes para entender como a forma e o tamanho são determinados, e como eles estão adaptados aos ambientes bióticos e abióticos. Fêmeas são mais pesadas que machos (acima de 1,75 g). Em habitat diferentes, o tórax e o abdômen das fêmeas é maior em 100 por cento das 11 espécies analisadas, a massa total 92 por cento, e a massa de asas 75 por cento. Espécies cujas fêmeas tiveram a maior massa também mostraram maior deslocamento e dispersão nas suas áreas de vida. Em habitats mais perturbados pelo homem ou com menor cobertura vegetal, o aumento no tamanho do tórax é significativamente maior que o das asas, tanto para os machos como para as fêmeas. Nos machos, o aumento de abdômen não é significativamente maior que o das asas, em todos os locais investigados. Porém, nas fêmeas o aumento do abdômen é significativo em todas os locais. A análise de agrupamento mostra dois grandes grupos caracterizados por espécies com tamanhos e proporções semelhantes. Um grupo com espécies que ocupam ambientes com umidade alta, e outro (mais heterogêneo) que ocupa habitats alterados pela ação do homem. O primeiro mostra duas subdivisões distinguidas pelo tipo de exibição que machos apresentam às fêmeas.

Reproductive aspects of the mosquito Culex quinquefasciatus (Diptera:Culicidae) infected with Wuchereria bancrofti (Spirurida: Onchocercidae)

This study reports on the relationship between Wuchereria bancrofti infection and female body size, intake of blood and fecundity in the mosquito Culex quinquefasciatus, vector of this filarial parasite in Recife (Brazil). Adults from field collected larvae were infected via a membrane feeding procedure, using blood with parasitaemia ranging from 724-6,000 mf/ml. A positive correlation was observed between mosquito size (measured by wing length) and egg production in uninfected females. However, this relationship did not exist in W. bancrofti infected mosquitoes. This change is unlikely to be the result of changes in blood ingestion as no significant difference was found when infected and uninfected females were compared. Variation in egg production observed between trials could not be associated with parasite density in the blood. These results suggest infection with W. bancrofti may disrupt the relationship between mosquito size and egg production during the first gonotrophic cycle of C. quinquefasciatus such that fecundity is sometimes reduced. However, this overall affect is variable and many groups of mosquitoes do not respond in this way

Geometric morphometric differences between Panstrongylus geniculatus from field and laboratory

The finding of Panstrongylus geniculatus nymphs inside a house in northeastern Antioquia, Colombia, and the reports related to their increasing presence in homes suggest the need for surveillance methods for monitoring the invasion processes. We analyzed the morphological differences between a wild population and its laboratory descendants, using the techniques of geometric morphometry, with the idea that such differences might parallel those between sylvatic and synanthropic populations. The analyses over five generations showed differences in size but not in shape. Head size and wing size were both reduced from sylvatic to laboratory populations, but the decrease in head size occurred only up to the second generation while the decrease in wing size proceeded up to the fifth generation. In contrast, although a decrease in sexual size dimorphism has been proposed as a marker of colonization in human dwellings, we did not detect any significant loss of dimorphism between sexes of P. geniculatus over the five generations studied. We conclude that size changes may have a physiological origin in response to a change of ecotopes, but more than five generations may be required for the expression of permanent morphological markers of human dwellings colonization