Gislene L. Gonçalves

A new Brazilian Passiflora leafminer: Spinivalva gaucha, gen. n., sp. n. (Lepidoptera, Gracillariidae, Gracillariinae), the first gracillariid without a sap-feeding instar.

Abstract Male, female, pupa, larva and egg of a new genus and species of Gracillariidae (Gracillariinae), Spinivalva gaucha Moreira and Vargas from southern Brazil are described and illustrated with the aid of optical and scanning electron microscopy. A preliminary analysis of mitochondrial DNA sequences including members of related lineages is also provided. The immature stages are associated with Passiflora actinia, Passiflora misera and Passiflora suberosa (Passifloraceae), and build mines on the adaxial leaf surface. Initially the mines are serpentine in shape, but later in larval ontogeny become a blotch type. Although the larvae are hypermetamorphic as in other Gracillariidae, there is no sap-feeding instar in Spinivalva gaucha; the larva feeds on the palisade parenchyma, thus producing granular frass during all instars. Pupation occurs outside the mine; prior to pupating, the larva excretes numerous bubbles that are placed in rows on the lateral margins of the cocoon external surface. This is the second genus of gracillariid moth described for the Atlantic Rain Forest, and the second gracillariid species known to be associated with Passifloraceae.

Prediction of the Damage-Associated Non- Synonymous Single Nucleotide Polymorphisms in the Human MC1R Gene

The melanocortin 1 receptor (MC1R) is involved in the control of melanogenesis. Polymorphisms in this gene have been associated with variation in skin and hair color and with elevated risk for the development of melanoma. Here we used 11 computational tools based on different approaches to predict the damage-associated non-synonymous single nucleotide polymorphisms (nsSNPs) in the coding region of the human MC1R gene. Among the 92 nsSNPs arranged according to the predictions 62% were classified as damaging in more than five tools. The classification was significantly correlated with the scores of two consensus programs. Alleles associated with the red hair color (RHC) phenotype and with the risk of melanoma were examined. The R variants D84E, R142H, R151C, I155T, R160W and D294H were classified as damaging by the majority of the tools while the r variants V60L, V92M and R163Q have been predicted as neutral in most of the programs The combination of the prediction tools results in 14 nsSNPs indicated as the most damaging mutations in MC1R (L48P, R67W, H70Y, P72L, S83P, R151H, S172I, L206P, T242I, G255R, P256S, C273Y, C289R and R306H); C273Y showed to be highly damaging in SIFT, Polyphen-2, MutPred, PANTHER and PROVEAN scores. The computational analysis proved capable of identifying the potentially damaging nsSNPs in MC1R, which are candidates for further laboratory studies of the functional and pharmacological significance of the alterations in the receptor and the phenotypic outcomes.

Larval feeding choices in heliconians: induced preferences are not constrained by performance and host plant phylogeny

Induction of feeding preference, a special type of learning found in the larval stage of herbivorous insects, may be a link between evolutionary changes in host selection behaviour, variation in diet breadth and evolution of host races. However, it has been argued that the phenomenon may be phylogenetically constrained in oligophagous species, which in general use closely related host plants. Also, induction might be tied to performance, being more likely to occur in host plants that are more suitable for larval feeding. If so, induction would be less likely to promote increase in diet breadth. By conducting reciprocal rearing and using leaf disks and double-choice feeding tests, we explored the influence of these factors on the induction of larval feeding preference in the first and last instars of two oligophagous heliconian butterflies ( Heliconius erato (Linnaeus) and Heliconius ethilla Godart) in relation to the main (five) passion vine species used as host plants by heliconians in southern Brazil. We also determined the effects of host plants on survivorship, growth rates and size attained in the adult stage for both butterfly species. For comparison, we also carried out a phylogenetic analysis of these plants, based on DNA sequencing. For both heliconian species, we found that feeding preference could be induced for most host plant species tested, in the fifth instar in particular, suggesting that habituation is involved in such cases. There was no indication of the existence of host plant phylogenetic constraints. We also found positive responses for induction on plants that supported poor larval performance. We conclude that induction of larval feeding preference in these cases was not limited by phylogenetic relatedness among the hosts or by their ability to support larval performance, which varies in spatial distribution and abundance in the area. We discuss the possible evolutionary consequences of this behavioural phenomenon in these heliconian butterflies.

DNA Barcoding of Sigmodontine Rodents: Identifying Wildlife Reservoirs of Zoonoses

Species identification through DNA barcoding is a tool to be added to taxonomic procedures, once it has been validated. Applying barcoding techniques in public health would aid in the identification and correct delimitation of the distribution of rodents from the subfamily Sigmodontinae. These rodents are reservoirs of etiological agents of zoonoses including arenaviruses, hantaviruses, Chagas disease and leishmaniasis. In this study we compared distance-based and probabilistic phylogenetic inference methods to evaluate the performance of cytochrome c oxidase subunit I (COI) in sigmodontine identification. A total of 130 sequences from 21 field-trapped species (13 genera), mainly from southern Brazil, were generated and analyzed, together with 58 GenBank sequences (24 species; 10 genera). Preliminary analysis revealed a 9.5% rate of misidentifications in the field, mainly of juveniles, which were reclassified after examination of external morphological characters and chromosome numbers. Distance and model-based methods of tree reconstruction retrieved similar topologies and monophyly for most species. Kernel density estimation of the distance distribution showed a clear barcoding gap with overlapping of intraspecific and interspecific densities < 1% and 21 species with mean intraspecific distance < 2%. Five species that are reservoirs of hantaviruses could be identified through DNA barcodes. Additionally, we provide information for the description of a putative new species, as well as the first COI sequence of the recently described genus Drymoreomys. The data also indicated an expansion of the distribution of Calomys tener. We emphasize that DNA barcoding should be used in combination with other taxonomic and systematic procedures in an integrative framework and based on properly identified museum collections, to improve identification procedures, especially in epidemiological surveillance and ecological assessments.

Sequence variation in the melanocortin-1 receptor (MC1R) pigmentation gene and its role in the cryptic coloration of two South American sand lizards

In reptiles, dorsal body darkness often varies with substrate color or temperature environment, and is generally presumed to be an adaptation for crypsis or thermoregulation. However, the genetic basis of pigmentation is poorly known in this group. In this study we analyzed the coding region of the melanocortin-1-receptor (MC1R) gene, and therefore its role underlying the dorsal color variation in two sympatric species of sand lizards (Liolaemus) that inhabit the southeastern coast of South America: L. occipitalis and L. arambarensis. The first is light-colored and occupies aeolic pale sand dunes, while the second is brownish and lives in a darker sandy habitat. We sequenced 630 base pairs of MC1R in both species. In total, 12 nucleotide polymorphisms were observed, and four amino acid replacement sites, but none of them could be associated with a color pattern. Comparative analysis indicated that these taxa are monomorphic for amino acid sites that were previously identified as functionally important in other reptiles. Thus, our results indicate that MC1R is not involved in the pigmentation pattern observed in Liolaemus lizards. Therefore, structural differences in other genes, such as ASIP, or variation in regulatory regions of MC1R may be responsible for this variation. Alternatively, the phenotypic differences observed might be a consequence of non-genetic factors, such as thermoregulatory mechanisms.

Mitochondrial and nuclear DNA analyses reveal population differentiation in Brazilian Creole sheep.

Using ND5 sequences from mtDNA and 10 nuclear markers, we investigated the genetic differentiation of two South American Creole sheep phenotypes that historically have been bred in different biomes in southern Brazil. In total, 18 unique mtDNA haplotypes were detected, none of which was shared between the two phenotypes. Bayesian analysis also indicated two different groups (k = 2). Thus, these varieties are supported as being genotypically distinct. This situation could have resulted either from geographical isolation, associated with differences in the cultural habits of sheep farmers and in the way that flocks were managed, or more likely, from the introduction of different stocks four centuries ago.

INTRASPECIFIC VARIATION AND GENETIC DIFFERENTIATION OF THE COLLARED TUCO-TUCO (CTENOMYS TORQUATUS) IN SOUTHERN BRAZIL

Abstract The collared tuco-tuco, Ctenomys torquatus Lichtenstein, 1830 (Ctenomyidae), is a subterranean rodent that occurs in grassland habitats of southern Brazil and northern Uruguay. A population of collared tuco-tucos located in Alegrete Municipality, Rio Grande do Sul State, Brazil, has been proposed as a potential new species because of their remarkable differences in chromosome number and their unique patterns of pelage coloration. The aim of this work was to evaluate the degree of genetic differentiation of this population using mitochondrial DNA (mtDNA) sequences and nuclear microsatellite markers, and to describe spatial patterns of genetic diversity for the other 5 populations of C. torquatus in southern Brazil, focusing on patterns of intra- and interpopulation variation in coat color. The analysis of 1,110 base pairs of the mtDNA cytochrome-b (Cytb) gene and 9 nuclear microsatellite loci revealed 7 haplotypes (n  =  65) and 48 alleles (n  =  70), respectively. Genetic diversity was moderately low within populations (HE  =  0.40–0.56), and significantly partitioned among locations (RST  =  0.21; P < 0.01). Analysis of the microsatellite data suggested that genetic differentiation is consistent with a simple model of isolation by distance (r  =  0.56, P < 0.05), and that the population is in equilibrium between gene flow and local genetic drift. The partially reconstructed phylogeny revealed that the haplotypes derived from the Alegrete population were not reciprocally monophyletic, and that there was a lack of structure for coat color and karyotype variation. Thus, the individuals from the Alegrete population fall within the range of variation for C. torquatus, and should not be considered a new species. We suggest that they be considered a local, specialized lineage that could be treated and managed from a conservation perspective as a Management Unit.

Revalidation Of Oliera Brèthes (Lepidoptera: Cecidosidae) Based On A Redescription Of O. Argentinana And Dna Analysis Of Neotropical Cecidosids

Larvae of Oliera argentinana, Brèthes 1916 (Lepidoptera: Cecidosidae) were rediscovered inducing spindle-shaped galls enclosed within swollen stems of Schinus (Anacardiaceae) in central Argentina and Rio Grande do Sul, the southernmost state of Brazil. Male, female, immature stages, and plant galls of O. argentinana are redescribed, using optical and scanning electron microscopy. The genus Oliera Brèthes, 1916, previously a junior synonym of Cecidoses Curtis, 1835, is revalidated, by comparing morphological characteristics within the family and through an analysis of mitochondrial (COI) DNA sequences, including putative members of the four Neotropical cecidosid genera. Information on preliminary Cecidosidae phylogeny and taxonomy is also provided.

A new species of swamp rat of the genus Scapteromys Waterhouse, 1837 (Rodentia: Sigmodontinae) endemic to Araucaria angustifolia Forest in Southern Brazil

A new species of swamp rat of the genus Scapteromys from the Meridional Plateau of Southern Brazil is described. Morphological, molecular, and karyological analysis support the recognition of the new species, distinct from S. aquaticus and S. tumidus. Scapteromys sp. nov. is significantly smaller than the congeneric taxa considering most of the external and craniometric measurements and the pelage is conspicuously grayer and darker. It can be distinguished from S. tumidus by the laterally extended thenar pad of the manus and the parallel edges of the hamular process of the pterygoid, and from S. aquaticus by a grayer and darker pelage and smaller values of most external and craniometric measurements. Karyological analysis indicated a difference in chromosome numbers across the distributional range: 2n=34 and 2n=36. A total of 11 haplotypes were found along the range of the new species within the biogeographic province of Araucaria angustifolia Forest. Strongly supported substructure was found within the new taxon, resulting in two reciprocally monophyletic clades.

Functional New World monkey oxytocin forms elicit an altered signaling profile and promotes parental care in rats

Significance Several forms of the oxytocin neurohormone have been found in New World monkeys (NWMs). Previous research has suggested an association between these forms and behaviors typical of this primate branch, including paternal care and monogamy. Our study provides genetic, pharmacological, behavioral, and in silico evidence supporting this connection. Rats treated intranasally with two NWM oxytocin variants showed an increase in some parental care behaviors. The same two variants were found to elicit different cell-signaling profiles in cell-based assays compared with ancestral oxytocin. Our findings highlight how mutations in the OXT DNA sequence coding for a nonapeptide result in distinct signaling profiles that may be linked to the emergence of novel adaptive traits, in this case, paternal care and monogamy. The neurohormone oxytocin is a key player in the modulation of reproductive and social behavioral traits, such as parental care. Recently, a correlation between different forms of oxytocin and behavioral phenotypes has been described in the New World Monkeys (NWMs). Here, we demonstrate that, compared with the Leu8OXT found in most placental mammals, the Cebidae Pro8OXT and Saguinus Val3Pro8OXT taxon-specific variants act as equi-efficacious agonists for the Gq-dependent pathway but are weaker agonists for the β-arrestin engagement and subsequent endocytosis toward the oxytocin receptor (OXTR). Upon interaction with the AVPR1a, Pro8OXT and the common Leu8OXT yielded similar signaling profiles, being equally efficacious on Gq and β-arrestin, while Val3Pro8OXT showed reduced relative efficacy toward β-arrestin. Intranasal treatment with either of the variants increased maternal behavior and also promoted unusual paternal care in rats, as measured by pup-retrieval tests. We therefore suggest that Val3Pro8OXT and Pro8OXT are functional variants, which might have been evolutionarily co-opted as an essential part of the adaptive genetic repertoire that allowed the emergence of taxon-specific complex social behaviors, such as intense parental care in the Cebidae and the genus Saguinus.