Monitoring of Group C Rotavirus in Children With Acute Gastroenteritis in Brazil: An Emergent Epidemiological Issue After Rotavirus Vaccine?

Group C rotavirus (GpCRV) has a worldwide distribution; however, its epidemiology and ecology are still unclear. Evidence for a possible zoonotic role has been postulated recently for Brazilian children strains. The aim of this study was to monitor GpCRV in children <= 15 years with acute gastroenteritis during the 2007-2010 national Brazilian rotavirus surveillance, and to undertake the molecular characterization of the major VP6 capsid protein. A total of 3,019 fecal samples were first screened for Group A rotavirus (GpARV). A total of 2,205 GpARV ELISA negative samples were tested further for the presence of GpCRV by SDS-PAGE, electronic microscopy, and RT-PCR for the VP6 gene. The genetic diversity of GpCRV was carried out by sequencing the VP6 gene. GpARV and GpCRV infections were detected in 24.6% (742/3,019) and 0.3% (8/3,019), respectively. The GpCRV detection rate increased from 0.2% (1/422) in 2007 to 1% (7/708) in 2008, and GpCRV cases were not detected in 2009 and 2010. The phylogenetic analysis indicated that the strains belonged to the human lineage, and showed a genetic relationship with the GpCRV strain from Japan isolated in 2009. None of the study sequences was related closely to animal GpCRV strains. This study provides further evidence that GpCRV is a minor cause of acute childhood gastroenteritis in Brazil, and does not suggest that GpCRV may assume epidemiological importance in the future, even after the introduction of a GpARV vaccine. In addition, the molecular analyses of the GpCRV samples in this study do not support the zoonotic hypothesis. J. Med. Virol. 83: 1631-1636, 2011. (C) 2011 Wiley-Liss, Inc.

Mechanisms of reproductive isolation between an ant species of hybrid origin and one of its parents.

The establishment of new species by hybridization is difficult because it requires the development of reproductive isolation (RI) in sympatry to escape the homogenizing effects of gene flow from the parental species. Here we investigated the role of two pre- and two postzygotic mechanisms of RI in a system comprising two interdependent Pogonomyrmex harvester ant lineages (the H1 and H2 lineages) of hybrid origin and one of their parental species (P. rugosus). Similar to most other ants, P. rugosus is characterized by an environmental system of caste determination with female brood developing either into queens or workers depending on nongenetic factors. By contrast, there is a strong genetic component to caste determination in the H1 and H2 lineages because the developmental fate of female brood depends on the genetic origin of the parents, with interlineage eggs developing into workers and intralineage eggs developing into queens. The study of a mixed mating aggregation revealed strong differences in mating flight timing between P. rugosus and the two lineages as a first mechanism of RI. A second important prezygotic mechanism was assortative mating. Laboratory experiments also provided support for one of the two investigated mechanisms of postzygotic isolation. The majority of offspring produced from the few matings between P. rugosus and the lineages aborted at the egg stage. This hybrid inviability was under maternal influence, with hybrids produced by P. rugosus queens being always inviable whereas a small proportion of H2 lineage queens produced large numbers of adult hybrid offspring. Finally, we found no evidence that genetic caste determination acted as a second postzygotic mechanism reducing gene flow between P. rugosus and the H lineages. The few viable P. rugosus-H hybrids were not preferentially shunted into functionally sterile workers but developed into both workers and queens. Overall, these results reveal that the nearly complete (99.5%) RI between P. rugosus and the two hybrid lineages stems from the combination of two typical prezygotic mechanisms (mating time divergence and assortative mating) and one postzygotic mechanism (hybrid inviability).

Anthropogenic disturbance as a driver of microspatial and microhabitat segregation of cytotypes of Centaurea stoebe and cytotype interactions in secondary contact zones.

BACKGROUND AND AIMS: In a mixed-ploidy population, strong frequency-dependent mating will lead to the elimination of the less common cytotype, unless prezygotic barriers enhance assortative mating. However, such barriers favouring cytotype coexistence have only rarely been explored. Here, an assessment is made of the mechanisms involved in formation of mixed-ploidy populations and coexistence of diploid plants and their closely related allotetraploid derivates from the Centaurea stoebe complex (Asteraceae). METHODS: An investigation was made of microspatial and microhabitat distribution, life-history and fitness traits, flowering phenology, genetic relatedness of cytotypes and intercytotype gene flow (cpDNA and microsatellites) in six mixed-ploidy populations in Central Europe. KEY RESULTS: Diploids and tetraploids were genetically differentiated, thus corroborating the secondary origin of contact zones. The cytotypes were spatially segregated at all sites studied, with tetraploids colonizing preferentially drier and open microhabitats created by human-induced disturbances. Conversely, they were rare in more natural microsites and microsites with denser vegetation despite their superior persistence ability (polycarpic life cycle). The seed set of tetraploid plants was strongly influenced by their frequency in mixed-ploidy populations. Triploid hybrids originated from bidirectional hybridizations were extremely rare and almost completely sterile, indicating a strong postzygotic barrier between cytotypes. CONCLUSIONS: The findings suggest that tetraploids are later immigrants into already established diploid populations and that anthropogenic activities creating open niches favouring propagule introductions were the major factor shaping the non-random distribution and habitat segregation of cytotypes at fine spatial scale. Establishment and spread of tetraploids was further facilitated by their superior persistence through the perennial life cycle. The results highlight the importance of non-adaptive spatio-temporal processes in explaining microhabitat and microspatial segregation of cytotypes.

Valor econômico e impacto da seleção para precocidade reprodutiva de fêmeas na raça Nelore

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

As rodovias e as aves de rapina nocturnas relação entre a densidade, o habitat e a mortalidade

As estradas e tráfego inerente surgem como a criação antrópica mais conspícua e penetrante na paisagem natural, sendo considerados os principais agentes causadores de fragmentação e destruição de habitats, assim como representam um obstáculo físico sem precedentes, limitando as relações directas entre os indivíduos, por diminuição da frequência de dispersão e aumento da mortalidade por atropelamento, impedindo o fluxo natural de genes e suscitando o aumento de fenómenos de inbreeding e perda de heterozigotia. Todos os impactes deletérios associados às rodovias são claramente perceptíveis em vertebrados, onde as aves de rapinas nocturnas não são excepção. Uma vez que estas rapaces beneficiam das suas bermas e orlas, como locais de poiso, nidificação ou como corredores de dispersão através da paisagem, são frequentemente vítimas de mortalidade por atropelamento em estradas, sendo esta problemática considerada actualmente uma das mais recentes e importantes formas de mortalidade não natural em rapinas nocturnas e vinculada como um dos maiores problemas de conservação que afecta este grupo. Não obstante, esse mútuo efeito de atracção/repulsa das estradas a estas rapaces, as rodovias criam uma barreira específica que limita a dinâmica, comportamento e densidade populacional das espécies residentes, reconhecendo-se que o isolamento daí resultante, pode comprometer a viabilidade populacional a longo prazo, podendo mesmo conduzir a altos riscos de extinção das populações locais devido a efeitos estocásticos. Mediante esta problemática, este trabalho debruçou-se sobre um único objectivo principal: a avaliação do impacte das rodovias e do tráfego, na densidade das aves de rapina nocturnas. Este estudo foi efectuado na região Alentejana, abrangendo uma área de cariz tipicamente mediterrânico, delimitada pelas localidades de Montemor-o-Novo, Arraiolos e Évora, sendo seccionada por 143 quilómetros de estradas, divididas em autoestrada, rodovias com elevada e reduzida densidade de tráfego. A monitorização das rapinas nocturnas foi conduzida em dois anos amostrais (2005 e 2007), tendo sido focalizada sobretudo em duas espécies de Strigiformes, a Coruja­ do-mato Strix aluco e o Mocho-galego Athene noctua, recorrendo ao uso de playbacks com reprodução de vocalizações de indivíduos conspecíficos. Foram usadas 32 variáveis explicativas integradas em três grandes grupos: variáveis de estrada, métricas da paisagem, uso do solo, tendo sido analiticamente testadas, recorrendo à aplicação de Modelos Lineares Generalizados. Os principais resultados obtidos demonstram que as variáveis de estrada, aliadas à densidade de tráfego e ruído inerente à sua circulação, são provavelmente, responsáveis por um comportamento de repulsa das espécies de aves de rapina nocturnas em estudo, apresentando estas densidades mais elevadas longe de áreas antropicamente perturbadas e, portanto, de menor qualidade que se encontram adjacentes às rodovias. Todavia a presença de habitat favorável a estas rapaces é provavelmente o descritor com maior poder estatístico no que concerne à sua distribuição e densidade, sendo os montados densos e a presença de zonas agrícolas de sequeiro, positivamente correlacionadas, respectivamente com a densidade de casais reprodutores de Coruja-do-mato e Mocho-galego. Mediante a observação dos resultados será veemente a aplicação de medidas de mitigação específicas, que fundamentalmente considerem o afastamento dos efectivos populacionais longe das estradas e tráfego, conservando e assegurando as características estruturais, requisitos e qualidade dos habitats, de modo a incrementar e garantir a viabilidade e densidade das populações, fidelizando a territorialidade e permanência destas aves nestas áreas. Adicionalmente deverá investir-se na conectividade das manchas de paisagem fragmentada pelas rodovias, criando opções de conservação estratégicas, em zonas ecologicamente mais sensíveis, que não somente minimizem o efeito de repulsa reconhecido nestas aves, mas também os níveis de mortalidade por atropelamento, tornando os ecossistemas mais funcionais para a sobrevivência destes rapaces. ABSTRACT; Roads and traffic are the most conspicuous and pervasive human creation, being the great responsible for fragmentation and habitats destruction, reducing animal movement through landscape, which implies decrease of gene flow and loss of variability that can fragmented populations, thus reducing their sizes and densities. All deleterious impacts associated with roads are clearly visible in vertebrates, where owls aren't exception, being frequent victims of road mortality, since they can use roadside habitats and edges for hunting, nesting or dispersal corridors through the landscape, being nowadays one of the most recent and important causes of non­natural mortality in owls and has been recognized as one of the largest conservation problems affecting this group. However, the attractive and avoidance effect of roads and his edges on owls creates a barrier effect that limits dynamics, behaviour and breeding density of resident species, recognizing that possible isolation, could compromise populations survival, make them more vulnerable to high risks of local extinction due to stochastic effects. Despite that, several authors suggest that owls use roads to hunt, as marginal habitats, or for navigation corridors through the landscape ln this context, the major aims of this study was to verify if there are negative effects of roads on the density of owls, considering traffic as an influencing factor. This study was conducted in Alentejo, covering a typical Mediterranean area, including three main localities Montemor-o-Novo, Évora and Arraiolos, being sectioned by 143 km of roads, including highway, roads with high traffic density, and the remaining with low traffic density. The owl census was conducted in two sampling years (2005 and 2007) and was focused mainly on Little owl Athene noctua and Tawny Owl Strix aluco species, using the playback technique, with conspecific calls. We used 32 explanatory variables, mainly included in three groups: road variables, landscape metrics and land use, having been analytically tested, with application of Generalised Linear Models. The main results show that noisy roads with high traffic density are probably the most responsible for the avoidance behaviour of owls, under the study area, showing density depression near high anthropogenic disturbed areas adjacent to roads. However, the presence of habitat quality to these birds is probably the descriptor with greater statistical power, considering its distribution and density, with the dense oak woodland and croplands and arable lands, positively correlated, respectively with Tawny owl and Little owl density. ln consequence, the great conservation effort should be done in order to keep breeding populations away from roads and traffic, ensuring the structural features, requirements and quality of its habitats in order to enhance and ensure the viability and density of owl's populations in these areas. ln addition, it is important to invest in connectivity between roadside fragmented patches, creating strategic conservation options, in sensitive areas, which minimize the avoidance effect, recognized in owls, but also road-kill levels, making ecosystems more functional to survival of these top predators.

Genetic variability of a population of Aedes aegypti from Paraná, Brazil, using the mitochondrial ND4 gene

Genetic variability of a population of Aedes aegypti from Paraná, Brazil, using the mitochondrial ND4 gene. To analyze the genetic variability of populations of Aedes aegypti, 156 samples were collected from 10 municipalities in the state of Paraná, Brazil. A 311 base pairs (bp) region of the NADH dehydrogenase subunit 4 (ND4) mitochondrial gene was examined. An analysis of this fragment identified eight distinct haplotypes. The mean genetic diversity was high (h = 0.702; p = 0.01556). AMOVA analysis indicated that most of the variation (67%) occurred within populations and the F ST value (0.32996) was highly significant. F ST values were significant in most comparisons among cities. The isolation by distance was not significant (r = -0.1216 and p = 0, 7550), indicating that genetic distance is not related to geographic distance. Neighbor-joining analysis showed two genetically distinct groups within Paraná. The DNA polymorphism and AMOVA data indicate a decreased gene flow in populations from Paraná, which can result in increased vectorial competence.

Variability and Genetic Structure in Populations of Arrabidaea bilabiata (Sprague) Sandwith in the Amazonas State

Weeds in pastures can intoxicate animals, and Arrabidaea bilabiata is the most important species for herbivores in floodplain areas in the Amazon Basin. Genetic diversity studies in natural populations may contribute to the better understanding of the range of toxicity and the genetic variability organization in this species. The objective of this study was to assess the variability and genetic structure in six populations of A. bilabiata sampled in floodplain areas in three municipalities of the Amazonas State, from the AFLP markers analysis. AFLP markers were efficient to characterize the genetic variability of the 65 individuals analyzed. From four combinations of oligonucleotides, a total of 309 AFLP fragments was obtained, where 304 (98.38%) were polymorphic. By the dendrogram and Bayesian cluster analysis, there was a formation of two isolated groups, the first one comprising individuals from Autazes municipality and the second one comprising individuals from Itacoatiara and Parintins. However, depending on the method to define the most probable cluster number, there was a separation of the six populations, according to their geographical origin. Mantel test confirmed that geographically closer populations are more akin, although low gene flow (0.538) is observed among the sampled populations. The molecular analysis of variance found that 49.29% of the genetic variability are among individuals inside populations and 50.71% among the populations analyzed. The results indicate the possibility that isolated A. bilabiata populations contain plants with different toxicity levels and suggest a strong adaptability of the species.

Stocking may increase mitochondrial DNA diversity but fails to halt the decline of endangered Atlantic salmon populations

Over the last 50 years, Spanish Atlantic salmon (Salmo salar) populations have been in decline. In order to bolster these populations, rivers were stocked with fish of northern European origin during the period 1974-1996, probably also introducing the furunculosis-inducing pathogen, Aeromonas salmonicida. Here we assess the relative importance of processes influencing mitochondrial (mt)DNA variability in these populations from 1948 to 2002. Genetic material collected over this period from four rivers in northern Spain (Cantabria) was used to detect variability at the mtDNA ND1 gene. Before stocking, a single haplotype was found at high frequency (0.980). Following stocking, haplotype diversity (h) increased in all rivers (mean h before stocking was 0.041, and 0.245 afterwards). These increases were due principally to the dramatic increase in frequency of a previously very low frequency haplotype, reported at higher frequencies in northern European populations proximate to those used to stock Cantabrian rivers. Genetic structuring increased after stocking: among-river differentiation was low before stocking (1950s/1960s Phi(ST) = -0.00296-0.00284), increasing considerably at the height of stocking (1980s Phi(ST) = 0.18932) and decreasing post-stocking (1990s/2002 Phi(ST) = 0.04934-0.03852). Gene flow from stocked fish therefore seems to have had a substantial role in increasing mtDNA variability. Additionally, we found significant differentiation between individuals that had probably died from infectious disease and apparently healthy, angled fish, suggesting a possible role for pathogen-driven selection of mtDNA variation. Our results suggest that stocking with non-native fish may increase genetic diversity in the short term, but may not reverse population declines.

Genetic structure of the ornamental tetra fish species Piabucus melanostomus Holmberg, 1891 (CHARACIDAE, IGUANODECTINAE) in the Brazilian Pantanal wetlands inferred by mitochondrial DNA sequences

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Geoffroy's side-necked turtle [Phrynops geoffroanus (Schweigger, 1812), Testudines: Chelidae] as a model for evolutionary ecotoxicology: relationship between environmental contamination, conditions and genetic variability

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Contemporary and historic factors influence differently genetic differentiation and diversity in a tropical palm

Population genetics theory predicts loss in genetic variability because of drift and inbreeding in isolated plant populations; however, it has been argued that long-distance pollination and seed dispersal may be able to maintain gene flow, even in highly fragmented landscapes. We tested how historical effective population size, historical migration and contemporary landscape structure, such as forest cover, patch isolation and matrix resistance, affect genetic variability and differentiation of seedlings in a tropical palm (Euterpe edulis) in a human-modified rainforest. We sampled 16 sites within five landscapes in the Brazilian Atlantic forest and assessed genetic variability and differentiation using eight microsatellite loci. Using a model selection approach, none of the covariates explained the variation observed in inbreeding coefficients among populations. The variation in genetic diversity among sites was best explained by historical effective population size. Allelic richness was best explained by historical effective population size and matrix resistance, whereas genetic differentiation was explained by matrix resistance. Coalescence analysis revealed high historical migration between sites within landscapes and constant historical population sizes, showing that the genetic differentiation is most likely due to recent changes caused by habitat loss and fragmentation. Overall, recent landscape changes have a greater influence on among-population genetic variation than historical gene flow process. As immediate restoration actions in landscapes with low forest amount, the development of more permeable matrices to allow the movement of pollinators and seed dispersers may be an effective strategy to maintain microevolutionary processes.

Genetic variability and differentiation in the Mediterranean endemic starry ray Raja asterias (Delaroche, 1809)

The present study deal with the population structure and connectivity of the Mediterranean endemic starry ray Raja asterias (Delaroche, 1809) in the Western and Eastern Mediterranean basin. A panel of eight microsatellite loci which cross-amplify in Rajidae (El Nagar, 2010) was used to assess population connectivity and structure. Those aims were investigated by analyzing the genetic variation of 9 population sample for a total of 185 individuals collected during past scientific surveys (MEDITS, GRUND), commercial trawling and also directly at fish markets. The purpose of this thesis is to estimate the genetic divergence occurring between the Mediterranean populations and, in particular, to assess the presence of any barrier (geographic, hydrogeological and biological) to gene flow for this species. Different statistical approaches were performed to reach this aim evaluating both the genetic diversity (nucleotide diversity, allelic richness, observed and expected heterozygosity and Hardy-Weinberg equilibrium test) and the population differentiation patterns (pairwise Fst estimated and population structure analysis). The results obtained from the analysis of the microsatellite dataset suggest a geographic and genetic separation between the starry ray populations of the Mediterranean basin into three or four distinct groups: Western and Eastern Mediterranean basins and Sicilian coast always clustering as an independent group and Algeria which could be or not considered another separate group. The data were discussed from both an evolutionary and a conservation point of view and in relation to previous results obtained by the analysis of mitochondrial marker. A comparison with other Mediterranean demersal skate species was performed in order to better contextualise our results. Finally, our results could offer useful information to protect vulnerable species as R. asterias and developing effective conservation plans in the Mediterranean.

Mitochondrial DNA Variability Among Eight Tikuna Villages: Evidence for an Intratribal Genetic Heterogeneity Pattern

To study the genetic structure of the Tikuna tribe, four major Native American mitochondrial DNA (mtDNA) founder haplogroups were analyzed in 187 Amerindians from eight Tikuna villages located in the Brazilian Amazon. The central position of these villages in the continent makes them relevant for attempts to reconstruct population movements in South America. In this geographic region, there is particular concern regarding the genetic structure of the Tikuna tribe, formerly designated ""enigmatic"" due to its remarkable degree of intratribal homogeneity and the scarcity of private protein variants. In spite of its large population size and geographic distribution, the Tikuna tribe presents marked genetic and linguistic isolation. All individuals presented indigenous mtDNA haplogroups. An intratribal genetic heterogeneity pattern characterized by two highly homogeneous Tikuna groups that differ considerably from each other was observed. Such a finding was unexpected, since the Tikuna tribe is characterized by a social system that favors intratribal exogamy and patrilocality that would lead to a higher female migration rate and homogenization of the mtDNA gene pool. Demographic explosions and religious events, which significantly changed the sizes and compositions of many Tikuna villages, may be reflected in the genetic results presented here. Am J Phys Anthropol 140:526-531,2009. (C) 2009 Wiley-Liss, Inc

Genetic variability of Triatoma flavida and Triatoma bruneri (Hemiptera: Reduviidae) by RAPD-PCR technique

The Triatominae (Hemiptera:Reduviidae) contains the principal and potential Chagas disease vectors present in Mexico, Central America and South America. Triatoma flavida and T. bruneri are Cuban species. These species are closely related according to morphology and were considered synonyms until 1981, when they were separated on the grounds of external characters of the body and the morphology of male genitalia. The present study seeks to analyze genetic polymorphism of T. flavida and T. bruneri populations using RAPD techniques, and to assess the genetic relationship between these species. Ten random primers were used to evaluate the genetic variability among species using RAPD-PCR. The genetic flow among them was calculated. The dendrogram based on calculated Jaccard distances showed two clearly distinguishable clusters which coincided with the studied species. Within each species, moderate genetic differentiation (Fst 0.05-0.15) and migration rates (N > 1) were found among populations, that reveal gene flow and genetic homogeneity. Between species, the Fst value showed a high genetic differentiation and the migration rate was insufficient to maintain genetic homogeneity, and confirmed the absence of gene flow between them. Our results confirm the genetic variability among T. flavida and T. bruneri species.

Genetic variability and differentiation between populations of Rhodnius prolixus and R. pallescens, vectors of Chagas' Disease in Colombia

Enzyme polymorphism in Rhodnius prolixus and R. pallescens (Hemiptera, Reduviidae), principal vectors of Chagas' disease in Colombia, was analyzed using starch gel electrophoresis. Three geographic locations were sampled in order to determine gene flow between populations and to characterize intra- and interspecific differences. Of 25 enzymes assayed 10 were successfully resolved and then used to score the genetic variation. The enzymes PEPD, GPI, PGM and ICD were useful to differentiate these species and PGD, PGM and MDH distinguished between sylvatic and domiciliary populations of R. prolixus. Both polymorphism and heterozygosity indicated greater genetic variability in sylvatic habitats (H = 0.021) compared to domiciliary habitats (H = 0.006) in both species. Gene flow between sylvatic and domiciliary populations in R. prolixus was found to be minimal. This fact and the genetic distance between them suggest a process of genetic isolation in the domiciliary population.