The Andes as a semi-permeable geographical barrier: Genetic connectivity between structured populations in a widespread spider.

Geographical barriers like mountain ranges impede genetic exchange among populations, promoting diversification. The effectiveness of these barriers in limiting gene flow varies between lineages due to each species' dispersal modes and capacities. Our understanding of how the Andes orogeny contributes to species diversification comes from well-studied vertebrates and a few arthropods and plants, neglecting organisms unable to fly or walk long distances. Some arachnids, such as Gasteracantha cancriformis, have been hypothesized to disperse long distances via ballooning (i.e. using their silk to interact with the wind). Yet, we do not know how the environment and geography shape its genetic diversity. Therefore, we tested whether the Andes contributed to the diversification of G. cancriformis acting as an absolute or semi-permeable barrier to genetic connectivity between populations of this spider at opposite sides of the mountain range. We sampled thousands of loci across the distribution of the species and implemented population genetics, phylogenetic, and landscape genetic analyses. We identified two genetically distinct groups structured by the Central Andes, and a third less structured group in the Northern Andes that shares ancestry with the previous two. This structure is largely explained by the altitude along the Andes, which decreases in some regions, possibly facilitating cross-Andean dispersal and gene flow. Our findings support that altitude in the Andes plays a major role in structuring populations in South America, but the strength of this barrier can be overcome by organisms with long-distance dispersal modes together with altitudinal depressions.

Las barreras geográficas como las cordilleras montañosas impiden el intercambio genético entre poblaciones, promoviendo la diversificación. La efectividad de estas barreras para limitar el flujo genético varía entre linajes debido a los modos y capacidades de dispersión de cada especie. Nuestra comprensión de cómo la orogenia de los Andes contribuye a la diversificación de especies proviene de vertebrados y algunos artrópodos y plantas bien estudiados, descuidando a los organismos incapaces de volar o caminar grandes distancias. Se ha hipotetizado que algunas arañas, como Gasteracantha cancriformis, se dispersan a grandes distancias mediante la técnica de "ballooning" (es decir, utilizando su seda para interactuar con el viento). Sin embargo, no sabemos cómo el entorno y la geografía han dado forma a su diversidad genética. Por lo tanto, probamos si los Andes contribuyeron a la diversificación de G. cancriformis actuando como una barrera absoluta o permeable para la conectividad genética entre poblaciones de esta araña en lados opuestos de la cordillera. Muestreamos miles de loci a través de la distribución de la especie e implementamos análisis de genética de poblaciones, filogenéticos y de genética del paisaje. Identificamos dos grupos genéticamente distintos estructurados por los Andes Centrales, y un tercer grupo menos estructurado en los Andes del Norte que comparte ascendencia con los dos anteriores. Esta estructura se explica en gran medida por la altitud a lo largo de los Andes, que disminuye en algunas regiones, posiblemente facilitando la dispersión y el flujo genético a través de los Andes. Nuestros hallazgos apoyan que la altitud en los Andes juega un papel importante en la estructuración de las poblaciones en América del Sur, pero la fuerza de esta barrera puede ser superada por organismos con modos de dispersión a larga distancia junto con depresiones altitudinales.

Adaptive introgression of a visual preference gene.

Visual preferences are important drivers of mate choice and sexual selection, but little is known of how they evolve at the genetic level. In this study, we took advantage of the diversity of bright warning patterns displayed by Heliconius butterflies, which are also used during mate choice. Combining behavioral, population genomic, and expression analyses, we show that two Heliconius species have evolved the same preferences for red patterns by exchanging genetic material through hybridization. Neural expression of regucalcin1 correlates with visual preference across populations, and disruption of regucalcin1 with CRISPR-Cas9 impairs courtship toward conspecific females, providing a direct link between gene and behavior. Our results support a role for hybridization during behavioral evolution and show how visually guided behaviors contributing to adaptation and speciation are encoded within the genome.

Female researchers are under-represented in the Colombian science infrastructure.

Worldwide women have increased their participation in STEM, but we are still far from reaching gender parity. Although progress can be seen at the bachelor's and master's level, career advancement of women in research still faces substantial challenges leading to a 'leaky pipeline' phenomenon (i.e., the continuous decrease of women's participation at advanced career stages). Latin America exhibits encouraging rates of women participation in research, but the panorama varies across countries and stages in the academic ladder. This study focuses on women's participation in research in natural sciences in Colombia and investigates career progression, leadership roles, and funding rates by analyzing data on scholarships, grants, rankings, and academic positions. Overall, we found persistent gender imbalances throughout the research ecosystem that were significant using classical statistical analyses. First, although women constitute >50% graduates from bachelors in natural sciences, <40% of researchers in this field are female. Second, women win <30% of research grants, and in turn, their scientific productivity is 2X lower than that of men. Third, because of the less research funding and output women have, their promotion to senior positions in academic and research rankings is slower. In consequence, only ~25% of senior researchers and full professors are women. Fourth, the proportion of women leading research groups and mentoring young scientist in Colombia is <30%. Our study deepens our understanding of gender gaps in STEM research in Colombia, and provides information to design initiatives that effectively target gender disparities by focusing on key areas of intervention, and then gradually building up, rather than tackling structural inequities all at once.

Divergent warning patterns influence male and female mating behaviours in a tropical butterfly.

Traits under divergent ecological selection that also function during mating can be important in maintaining species boundaries. Few studies have considered mutual mate choice, where both males and females base mating decisions on the same trait. Wing colouration in Heliconius butterflies evolved as a warning signal but also functions as a mating cue. We investigated the contribution of visual preference to assortative mating in an aposematic butterfly Heliconius cydno in the context of reproductive isolation with its sympatric, visually distinct relative Heliconius melpomene. Heliconius cydno have conspicuous white bands on their forewings, whereas those of H. melpomene are red in colour. We predicted that both sexes of H. cydno contributed to assortative mating by exhibiting visual preference towards conspecific wing colouration. We analysed published and new data from preference experiments, in which males were presented with conspecific and H. melpomene females. We also recorded female responses and mating outcomes in choice experiments, involving conspecific males with either the original white or artificially painted red forewing bands. Both sexes of H. cydno responded more positively towards the conspecific colouration, and males strongly preferred females of its own colours. In contrast, male colouration did not predict mating outcomes in female choice experiments. As courtships are initiated by males in butterflies, our findings suggest that female visual preference might be of secondary importance in H. cydno. Our data also suggest that the contribution of visual preference to reproductive isolation might be unequal between H. cydno and its sympatric relative H. melpomene.

Parallel shifts in flight-height associated with altitude across incipient Heliconius species.

Vertical gradients in microclimate, resource availability, and interspecific interactions are thought to underly stratification patterns in tropical insect communities. However, only a few studies have explored the adaptive significance of vertical space use during the early stages of reproductive isolation. We analysed flight-height variation across speciation events in Heliconius butterflies, representing parallel colonizations of high-altitude forest. We measured flight-height in wild H. erato venus and H. chestertonii, parapatric lowland and mountain specialists, respectively, and found that H. chestertonii consistently flies at a lower height. By comparing our data to previously published results for the ecologically equivalent H. e. cyrbia (lowland) and H. himera (high altitude), we found that the species flying closest to the ground are those that recently colonized high-altitude forests. We show that these repeated trends largely result from shared patterns of ecological selection producing parallel trait-shifts in H. himera and H. chestertonii. Although our results imply a signature of local adaptation, we did not find an association between resource distribution and flight-height in H. e. venus and H. chestertonii. We discuss how this pattern may be explained by variations in forest structure and microclimate. Overall, our findings underscore the importance of behavioural adjustments during early divergence mediated by altitude-shifts.

Parallel evolution of behavior, physiology, and life history associated with altitudinal shifts in forest type in Heliconius butterflies.

Parallel evolution of morphological traits is widely reported, providing evidence for the role of local conditions in driving adaptive divergence. Comparatively, fewer studies have tested for parallelism in behavior, and it is less clear to what extent heritable behavioral shifts contribute to adaptive divergence. We exploit repeated incipient speciation across altitudinal gradients to explore behavior and physiology in Heliconius butterflies adapted to high-elevation. We performed common garden experiments with H. chestertonii, a high-altitude specialist from the Colombian Cordillera Occidental, and H. erato venus, a low-elevation proxy for the ancestral population, and compared our results to existing data for an equivalent Ecuadorian taxa-pair. Using broad-scale climatic data, we show that both pairs diverge across similar ecological gradients, confirmed using localized data loggers in the ranges of H. chestertonii and H. e. venus. We further show that H. chestertonii and H. e. venus have divergent activity patterns, attributable to different responses to microclimate, and life histories. Finally, we provide evidence for parallelism in these traits with H. himera and H. e. cyrbia. We propose that this is a result of selection associated with independent colonizations of high-altitude forests, emphasizing the importance of heritable behavioral and physiological adaptations during population divergence and speciation.

Geography shapes the microbial community in Heliconius butterflies.

Heliconius butterflies are an ideal organism for studying ecology, behavior, adaptation, and speciation. These butterflies can be found in various locations and habitats in Central and South America, where they encounter and interact with different sources of pollen, nectar, and host plants. However, there is limited knowledge on how geographic and habitat variations affect the microbiota of these insects, and whether microbial associates play a role in their ability to exploit different habitats. To date, research on the microbial communities associated with Heliconius has mainly focused on host phylogenetic signal in microbiomes or microbiome characterization in specific communities of butterflies. In this study, we characterized the microbiomes of several species and populations of Heliconius from distant locations that represent contrasting environments. We found that the microbiota of different Heliconius species is taxonomically similar but vary in abundance. Notably, this variation is associated with a major geographic barrier-the Central Cordillera of Colombia. Additionally, we confirmed that this microbiota is not associated with pollen-feeding. Therefore, it seems likely that geography shapes the abundance of microbiota that the butterfly carries, but not the taxonomic diversity of the microbial community. Based on the current evidence, the bacterial microbiota associated with Heliconius does not appear to play a beneficial role for these butterflies.

Does sexual conflict contribute to the evolution of novel warning patterns?

Why warning patterns are so diverse is an enduring evolutionary puzzle. Because predators associate particular patterns with unpleasant experiences, an individual's predation risk should decrease as the local density of its warning pattern increases, promoting pattern monomorphism. Distasteful Heliconius butterflies are known for their diversity of warning patterns. Here, we explore whether interlocus sexual conflict can contribute to their diversification. Male Heliconius use warning patterns as mating cues, but mated females may suffer costs if this leads to disturbance, favouring novel patterns. Using simulations, we show that under our model conditions drift alone is unlikely to cause pattern diversification, but that sexual conflict can assist such a process. We also find that genetic architecture influences the evolution of male preferences, which track changes in warning pattern due to sexual selection. When male attraction imposes costs on females, this affects the speed at which novel pattern alleles increase. In two experiments, females laid fewer eggs with males present. However, although males in one experiment showed less interest in females with manipulated patterns, we found no evidence that female colouration mitigates sex-specific costs. Overall, male attraction to conspecific warning patterns may impose an unrecognized cost on Heliconius females, but further work is required to determine this experimentally.

Repair Bond Strength of Aged Composite Resins Using Different Surface Treatment Protocols.

This study evaluated shear bond strength (SBS) of thermally aged composite resins repaired using different surface protocols. Four-hundred composite resin samples were made using the following materials (100 samples per material): Filtek Z350XT (FXT); Spectra Smart (SSM); IPS Empress Direct (EDI); and Forma (FOR). Each group's samples were then divided into 10 groups (n = 10 samples per group): G1: no surface treatment; G2: phosphoric acid-etching + universal-adhesive (PU); G3: surface roughening + PU (RPU); G4: RPU + silane (RPSU); G5: surface roughening + hydrofluoric acid-etching + universal adhesive (RHU); G6: RHU + silane (RHSU); G7: dry sandblast + PU (DsPU); G8: DsPU + silane (DsPSU); G9: wet sandblast + PU (WsPU); and G10: WsPU + silane (WsPSU). G1 was freshly repaired, and G2 to G10 were thermally aged before repair. Specimens were tested for SBS, and the failure type was observed with a magnifying loupe. Representative images were obtained using a scanning electronic microscope. Data were analyzed by two-way analysis of variance and Tukey post hoc tests (P = .05). Differences were detected among different surface treatments and among different composite resins with equal surface treatments (P < .05). SBS means ranged from 10.48 (FOR:G2) to 20.70 (FXT:G7). The highest SBS values were seen in G7 to G10 (P > .05), while lowest values were generally observed for G2. G1 showed higher results compared to G2 (P < .05), except for EDI (P > .05). Most failures corresponded with cohesive type. In general, thermally aged composite resin presented a decreased repair bond strength potential when no additional surface treatment was applied. Sandblasting improved the SBS of repaired aged composite resins.

Comparison of a resin-based sealant with a nano-filled flowable resin composite on sealing performance of marginal defects in resin composites restorations: a 36-months clinical evaluation.

OBJECTIVES: Our goal was to evaluate the clinical behavior of resin-based composite (RBC) restorations with sealed marginal defects using nano-filled flowable RBCs (FRS) compared with resin-based sealant (RBS); this work used marginal adaptation, marginal staining, and secondary caries according to the World Dental Federation (FDI) criteria. MATERIALS AND METHODS: This was a prospective, randomized, double-blind, controlled trial. Fifty-four patients who met the inclusion criteria (older than 18 years old; with high cariogenic risk determined by Cariogram software; and restorations with marginal defects, 3 and 4 according to FDI criteria) were randomly divided into three groups. There were three defective RBC restorations per patient and were repaired (n = 162). The groups were RBS-marginal sealing using a resin-based sealant (Clinpro Sealant, 3 M ESPE, MN, USA) plus adhesive (Single Bond Universal, 3 M ESPE, MN, USA); FRS-sealing using flowable resin (Filtek Flow Z350XT, 3 M ESPE, MN, USA) plus adhesive (Single Bond Universal, 3 M ESPE, MN, USA); and control-no repair treatment. All procedures were performed under complete isolation. Evaluations were evaluated at 1-week post treatment (baseline) as well as at 18 and 36 months after treatment regarding marginal adaptation, marginal staining, and secondary caries according to FDI criteria. The data were analyzed using the Wilcoxon test (α = 0.05) to compare the differences in each treatment group at different evaluation times. RESULTS: Marginal adaptation of micro-repaired RBC restorations were seen in patients with a high risk of caries using flowable resin composite or resin-based sealants. There were differences (P < 0.001) when baseline was compared at 18 and 36 months. Marginal staining showed differences when baseline was compared to 18 months (P < 0.001) and 36 months (P = 0.001) for both treatments. Secondary caries parameters for RBS treatment showed differences when baseline was compared to 36 months (P = 0.025) and when 18 months was compared to 36 months (P = 0.046). CONCLUSIONS: Micro-repair of RBC restorations resulted in clinical deterioration of marginal adaptation and marginal staining. Nano-filled flowable resin composites were sealed on defective restorations; 3 and 4 FDI marginal defects have better clinical performance to prevent secondary caries than resin-based sealants after 36 months. CLINICAL RELEVANCE: Micro-repair with RBS does not seem to be an effective treatment to prevent secondary caries.

Light environment influences mating behaviours during the early stages of divergence in tropical butterflies.

Speciation is facilitated when traits under divergent selection also act as mating cues. Fluctuations in sensory conditions can alter signal perception independently of adaptation to the broader sensory environment, but how this fine-scale variation may constrain or promote behavioural isolation has received little attention. The warning patterns of Heliconius butterflies are under selection for aposematism and act as mating cues. Using computer vision, we extracted behavioural data from 1481 h of video footage, for 387 individuals. We show that the putative hybrid species H. heurippa and its close relative H. timareta linaresi differ in their response to divergent warning patterns, but that these differences are strengthened with increased local illuminance. Trials with live individuals reveal low-level assortative mating that is sufficiently explained by differences in visual attraction. Finally, results from hybrid butterflies are consistent with linkage between a major warning pattern gene and the corresponding behaviour, though the differences in behaviour we observe are unlikely to cause rapid reproductive isolation as predicted under a model of hybrid trait speciation. Overall, our results reveal that the contribution of ecological mating cues to reproductive isolation may depend on the immediate sensory conditions during which they are displayed to conspecifics.

Müllerian mimicry of a quantitative trait despite contrasting levels of genomic divergence and selection.

Hybrid zones, where distinct populations meet and interbreed, give insight into how differences between populations are maintained despite gene flow. Studying clines in genetic loci and adaptive traits across hybrid zones is a powerful method for understanding how selection drives differentiation within a single species, but can also be used to compare parallel divergence in different species responding to a common selective pressure. Here, we study parallel divergence of wing colouration in the butterflies Heliconius erato and H. melpomene, which are distantly related Müllerian mimics which show parallel geographic variation in both discrete variation in pigmentation, and quantitative variation in structural colour. Using geographic cline analysis, we show that clines in these traits are positioned in roughly the same geographic region for both species, which is consistent with direct selection for mimicry. However, the width of the clines varies markedly between species. This difference is explained in part by variation in the strength of selection acting on colour traits within each species, but may also be influenced by differences in the dispersal rate and total strength of selection against hybrids between the species. Genotyping-by-sequencing also revealed weaker population structure in H. melpomene, suggesting the hybrid zones may have evolved differently in each species, which may also contribute to the patterns of phenotypic divergence in this system. Overall, we conclude that multiple factors are needed to explain patterns of clinal variation within and between these species, although mimicry has probably played a central role.

Interplay between Developmental Flexibility and Determinism in the Evolution of Mimetic Heliconius Wing Patterns.

To what extent can we predict how evolution occurs? Do genetic architectures and developmental processes canalize the evolution of similar outcomes in a predictable manner? Or do historical contingencies impose alternative pathways to answer the same challenge? Examples of Müllerian mimicry between distantly related butterfly species provide natural replicates of evolution, allowing us to test whether identical wing patterns followed parallel or novel trajectories. Here, we explore the role that the signaling ligand WntA plays in generating mimetic wing patterns in Heliconius butterflies, a group with extraordinary mimicry-related wing pattern diversity. The radiation is relatively young, and numerous cases of wing pattern mimicry have evolved within the last 2.5-4.5 Ma. WntA is an important target of natural selection and is one of four major effect loci that underlie much of the pattern variation in the group. We used CRISPR/Cas9 targeted mutagenesis to generate WntA-deficient wings in 12 species and a further 10 intraspecific variants, including three co-mimetic pairs. In all tested butterflies, WntA knockouts affect pattern broadly and cause a shift among every possible scale cell type. Interestingly, the co-mimics lacking WntA were very different, suggesting that the gene networks that pattern a wing have diverged considerably among different lineages. Thus, although natural selection channeled phenotypic convergence, divergent developmental contexts between the two major Heliconius lineages opened different developmental routes to evolve resemblance. Consequently, even under very deterministic evolutionary scenarios, our results underscore a surprising unpredictability in the developmental paths underlying convergence in a recent radiation.

Volumetric polymerization shrinkage and its comparison to internal adaptation in bulk fill and conventional composites: A µCT and OCT in vitro analysis.

OBJECTIVE: To quantify the volumetric polymerization shrinkage (VPS) of different conventional and bulk fill resin composites, through micro-computed tomography (µCT), and qualitative comparison of gap formation through optical coherence tomography (OCT). METHODS: Box-shaped class I cavities were prepared in 30 third-molars and divided into 5 groups (n=6): G1- Filtek Z100 (Z100); G2- Tetric Evoceram Bulk Fill (TEC); G3- Tetric EvoFlow Bulk fill (TEF); G4- Filtek Bulk fill (FBU); and G5- Filtek Bulk fill Flowable (FBF). All groups were treated with Adper Single Bond Plus adhesive and light cured (Bluephase 20i). Each tooth was scanned three times using a µCT apparatus: after cavity preparation (empty scan); after cavity filling (uncured scan) and after light curing of the restorations (cured scan). The µCT images were imported into a three-dimensional rendering software, and volumetric polymerization shrinkage percentage was calculated (%) for each sample. In the same images, interfacial gaps in the pulpal floor were qualitatively evaluated. After µCT evaluation, the pulpal floor from each tooth was polished until a thin tooth structure was obtained and OCT images were obtained by scanning the pulpal portion. Gap formation was observed and qualitatively compared to the µCT images. RESULTS: VPS means ranged from 2.31 to 3.96% for the studied resin composites. The bulk fill materials, either high viscosity or flowable, were not statistically different from each other (p>0.05). The conventional resin composite Z100 presented statistically higher VPS than both high viscosity bulk fill materials studied (p<0.05), although it was statistically similar to the flowable bulk fill materials studied (p>0.05). Both µCT and OCT methodologies enabled gap formation visualization, and images from both technologies could be associated. Gap formation was mostly observed for G1-Z100, G4-FBU, and G5-FBF. VPS% and pulpal gap formation could not be completely associated with each other for all groups and samples. Voids were observed in most of the resin composite fillings, and most VPS were observed in the occlusal area of the samples. SIGNIFICANCE: Volumetric polymerization shrinkage was material-dependent, although bulk fill materials did not differ from each other. Both µCT and OCT enabled interfacial pulpal gap formation visualization. VPS and gap formation cannot be completely associated with one another.

Taxonomic reassessment of the genus Dichotomius (Coleoptera: Scarabaeinae) through integrative taxonomy.

Dung beetles of the subfamily Scarabaeinae are widely recognised as important providers of multiple ecosystem services and are currently experiencing revisions that have improved our understanding of higher-level relationships in the subfamily. However, the study of phylogenetic relationships at the level of genus or species is still lagging behind. In this study we investigated the New World beetle genus Dichotomius, one of the richest within the New World Scarabaeinae, using the most comprehensive molecular and morphological dataset for the genus to date (in terms of number of species and individuals). Besides evaluating phylogenetic relationships, we also assessed species delimitation through a novel Bayesian approach (iBPP) that enables morphological and molecular data to be combined. Our findings support the monophyly of the genus Dichotomius but not that of the subgenera Selenocopris and Dichotomius sensu stricto (s.s). Also, our results do not support the recent synonymy of Selenocopris with Luederwaldtinia. Some species-groups within the genus were recovered, and seem associated with elevational distribution. Our species delimitation analyses were largely congruent irrespective of the set of parameters applied, but the most robust results were obtained when molecular and morphological data were combined. Although our current sampling and analyses were not powerful enough to make definite interpretations on the validity of all species evaluated, we can confidently recognise D. nisus, D. belus and D. mamillatus as valid and well differentiated species. Overall, our study provides new insights into the phylogenetic relationships and classification of dung beetles and has broad implications for their systematics and evolutionary analyses.

Taxonomical over splitting in the Rhodnius prolixus (Insecta: Hemiptera: Reduviidae) clade: Are R. taquarussuensis (da Rosa et al., 2017) and R. neglectus (Lent, 1954) the same species?

The use of subtle features as species diagnostic traits in taxa with high morphological similarity sometimes fails in discriminating intraspecific variation from interspecific differences, leading to an incorrect species delimitation. A clear assessment of species boundaries is particularly relevant in disease vector organisms in order to understand epidemiological and evolutionary processes that affect transmission capacity. Here, we assess the validity of the recently described Rhodnius taquarussuensis (da Rosa et al., 2017) using interspecific crosses and molecular markers. We did not detect differences in hatching rates in interspecific crosses between R. taquarussuensis and R. neglectus (Lent, 1954). Furthermore, genetic divergence and species delimitation analyses show that R. taquarussuensis is not an independent lineage in the R. prolixus group. These results suggest that R. taquarussuensis is a phenotypic form of R. neglectus instead of a distinct species. We would like to stress that different sources of evidence are needed to correctly delimit species. We consider this is an important step in understanding vectorial Chagas disease spread and transmission.

Gene flow and Andean uplift shape the diversification of Gasteracantha cancriformis (Araneae: Araneidae) in Northern South America.

The Andean uplift has played a major role in shaping the current Neotropical biodiversity. However, in arthropods other than butterflies, little is known about how this geographic barrier has impacted species historical diversification. Here, we examined the phylogeography of the widespread color polymorphic spider Gasteracantha cancriformis to evaluate the effect of the northern Andean uplift on its divergence and assess whether its diversification occurred in the presence of gene flow. We inferred phylogenetic relationships and divergence times in G. cancriformis using mitochondrial and nuclear data from 105 individuals in northern South America. Genetic diversity, divergence, and population structure were quantified. We also compared multiple demographic scenarios for this species using a model-based approach (phrapl) to determine divergence with or without gene flow. At last, we evaluated the association between genetic variation and color polymorphism. Both nuclear and mitochondrial data supported two well-differentiated clades, which correspond to populations occurring on opposite sides of the Eastern cordillera of the Colombian Andes. The final uplift of this cordillera was identified as the most likely force that shaped the diversification of G. cancriformis in northern South America, resulting in a cis- and trans-Andean phylogeographic structure for the species. We also found shared genetic variation between the cis- and trans-Andean clades, which is better explained by a scenario of historical divergence in the face of gene flow. This has been likely facilitated by the presence of low-elevation passes across the Eastern Colombian cordillera. Our work constitutes the first example in which the Andean uplift coupled with gene flow influenced the evolutionary history of an arachnid lineage.

Análise comparativa e caracterização das bolhas de ar em dois modelos de técnicas restauradoras: incremental e bulk fill / Comparative analysis of voids quantity in two restorative techniques: incremental and bulk fill

Os materiais restauradores estão evoluindo rapidamente e novas técnicas de inserção do compósito na cavidade restauradora estão sendo propostos. Dependendo da técnica, poderá ocorrer maior ou menor formação de bolhas na restauração final. A incorporação de bolhas entre as camadas do material restaurador pode gerar uma maior degradação da restauração em virtude da menor obtenção de propriedades físico-mecânicas do material. Assim, a necessidade de estudos sobre a prevalência de bolhas em restaurações confeccionadas com diferentes técnicas de inserção se faz necessária para a melhor compreensão do uso e do manuseio dos materiais. O objetivo desse trabalho foi analisar a ocorrência de bolhas e o volume de espaços presentes no interior de restaurações com resina composta, inserida na cavidade dental através das técnicas restauradoras: incremental e bulk fill. Quarenta e oito dentes molares humanos (n=24, duas restaurações por dente) foram selecionados e, nestes, foram confeccionados dois preparos por dente, um em cada região proximal, com profundidade de 4mm, largura de 4mm e distância axio-proximal de 2mm. Posteriormente, foram restaurados de acordo com os grupos: FiltekTM One Bulk Fill Restorative (FOB; 3M Oral Care); FiltekTM Z350 XT (FXT; 3M Oral care); FiltekTM Bulk Fill Flowable Restorative (FBF+FXT; 3M Oral care) e FiltekTM Z350 XT Flow (FF+FXT; 3M Oral care) . As resinas fluidas foram cobertas com a resina FiltekTM Z350 XT (3M Oral Care). Após 24 horas, os dentes restaurados foram submetidos à ciclagem térmica (5.000 ciclos) e posteriormente foi realizada a Tomografia de Coerência Ótica (OCT). Através da análise das imagens obtidas pelo OCT foram medidos os valores de quantidade de bolhas e porcentagem de espaços presentes nas restaurações. Os dados da presença de bolhas foram analisados estatisticamente com o teste Qui-Quadrado, os dados de número de bolhas e porcentagem de espaços obtidos foram analisados com o teste de Kruskall Wallis e a comparação entre os grupos foi feita pelo método de Dunn , com nível de significância de 5%. FF+FXT, apresentou bolhas em todas as restaurações e foi significativamente maior quando comparado a FXT (p=0,015). FF+FXT apresentou quantidade significativamente maior de bolhas (p<0,05) quando comparada com os outros três grupos. FBF+FXT apresentou a maior porcentagem de porosidade, devido ao maior tamanho de bolhas, porém esta porcentagem foi significantemente maior apenas quando comparada com FXT (p<0,05). Todos os grupos apresentaram bolhas. As técnicas restauradoras que apresentaram o pior desempenho, considerando o número de bolhas e a porcentagem de porosidade, foram aquelas realizadas com resina composta fluída. Não houve diferença estatisticamente significante quanto ao número de bolhas e à porcentagem de espaços entre as restaurações confeccionadas somente com resinas de alta viscosidade para as diferentes técnicas utilizadas neste estudo.

Male sex pheromone components in Heliconius butterflies released by the androconia affect female choice.

Sex-specific pheromones are known to play an important role in butterfly courtship, and may influence both individual reproductive success and reproductive isolation between species. Extensive ecological, behavioural and genetic studies of Heliconius butterflies have made a substantial contribution to our understanding of speciation. Male pheromones, although long suspected to play an important role, have received relatively little attention in this genus. Here, we combine morphological, chemical and behavioural analyses of male pheromones in the Neotropical butterfly Heliconius melpomene. First, we identify putative androconia that are specialized brush-like scales that lie within the shiny grey region of the male hindwing. We then describe putative male sex pheromone compounds, which are largely confined to the androconial region of the hindwing of mature males, but are absent in immature males and females. Finally, behavioural choice experiments reveal that females of H. melpomene, H. erato and H. timareta strongly discriminate against conspecific males which have their androconial region experimentally blocked. As well as demonstrating the importance of chemical signalling for female mate choice in Heliconius butterflies, the results describe structures involved in release of the pheromone and a list of potential male sex pheromone compounds.

The Scent Chemistry of Heliconius Wing Androconia.

Neotropical Heliconius butterflies are members of various mimicry rings characterized by diverse colour patterns. In the present study we investigated whether a similar diversity is observed in the chemistry of volatile compounds present in male wing androconia. Recent research has shown that these androconia are used during courting of females. Three to five wild-caught male Heliconius individuals of 17 species and subspecies were analyzed by GC/MS. Most of the identified compounds originate from common fatty acids precursors, including aldehydes, alcohols, acetates or esters preferentially with a C18 and C20 chain, together with some alkanes. The compounds occurred in species-specific mixtures or signatures. For example, octadecanal is characteristic for H. melpomene, but variation in composition between the individuals was observed. Cluster analysis of compound occurrence in individual bouquets and analyses based on biosynthetic motifs such as functional group, chain length, or basic carbon-backbone modification were used to reveal structural patterns. Mimetic pairs contain different scent bouquets, but also some compounds in common, whereas sympatric species, both mimetic and non-mimetic, have more distinct compound compositions. The compounds identified here may play a role in mate choice thus helping maintain species integrity in a butterfly genus characterized by pervasive interspecific gene flow.