Molecular diagnosis of the arowanas Osteoglossum ferreirai Kanazawa, 1966 and O. bicirrhossum (Cuvier, 1829) from the Orinoco and Amazon River basins

The arowanas, fishes of Gondwanan origin, are represented in South America by the genus Osteoglossum. All species were initially reported as being exclusive to the Amazon region, with O. ferreirai restricted to the Negro River basin and O. bicirrhosum to the Amazon and Essequibo Rivers basin. Starting in the mid 1970's it was reported that O. ferreirai also occurs in the Orinoco River basin. In all regions the arowanas assumed significant socio-economic importance due to their popularity in the international ornamental fish trade, leading to over-exploitation of both species in some areas. The Orinoco populations are particularly heavily exploited, and thus conservation and management measures are needed. Both depend on the clarification of taxonomic status, and phylogenetic distinctness of the Orinoco populations. With the goal of molecularly characterizing the two species of Osteoglossum, and comparing populations of Osteoglossum from the Orinoco and Amazon basins, we characterized individuals sampled from eight localities, one in the Orinoco River basin and seven in the Amazon River basin. We sampled 39 individuals, obtaining 1004 base pairs, of which 79 were synapomorphies. Genetic distance between the two species calculated using the HKY + G model of molecular evolution was 8.94%. Intraspecific distances ranged from 0.42% in O. bicirrhosum to 0.10% in O. ferreirai. The genetic characterization confirmed the taxonomic status of O. ferreirai in the Orinoco basin, and suggested that its distribution in the Orinoco basin is unlikely to be the result of vicariance or natural dispersal, but rather an anthropic introduction.

Os aruanãs, peixes de origem Gondwana, são representados na América do Sul pelo gênero Osteoglossum. Todas as espécies foram inicialmente reportadas como sendo exclusivamente da região Amazônica, com O. ferreirai restrito a bacia do Rio Negro e O. bicirrhosum para a bacia Amazônica e rio Essequibo. No meio dos anos 70 foi reportado que O. bicirrhosum também ocorre na bacia do rio Orinoco. Em todas as regiões os aruanãs são de significante importância socio-econômica devido a sua popularidade no comércio internacional de peixes ornamentais, levando a sobre-exploração de ambas as espécies em algumas áreas. As populações no rio Orinoco são particularmente muito exploradas, e assim medidas de conservação e manejo são necessárias. Ambas as medidas dependem de clarificações do estatus taxonômico, e distinções filogenéticas das populações do Orinoco. Com o objetivo de caracterizar molecularmente as duas espécies de Osteoglossum, e comparar as populações de Osteoglossum das bacias do Orinoco e Amazonas, nós caracterizamos indivíduos amostrados de oito localidades, uma da bacia do rio Orinoco e sete da bacia Amazônica. Nós amostramos 39 indivíduos, obtendo 1004 pares de bases, dos quais 97 foram sinapomorfias. A distância genética entre as duas espécies, calculadas usando-se o modelo HKY+G de evolução molecular foi de 8.94%. Distâncias intraespecíficas variaram de 0.42% em O. bicirrhosum a 0.10% em O. ferreirai. A caracterização genética confirmou o estatus taxonômico de O. ferreirai na bacia do rio Orinoco, e sugere que sua distribuição na bacia do rio Orinoco é improvavel de ser o resultado de um evento de vicariância ou dispersão natural, sendo melhor explicada como uma introdução antrópica.

Phylogeography and conservation genetics of the Amazonian freshwater stingray Paratrygon aiereba Müller & Henle, 1841 (Chondrichthyes: Potamotrygonidae)

The family Potamotrygonidae is monophyletic comprising three genera: Paratrygon Duméril, Potamotrygon Garman and Plesiotrygon Rosa, Castello & Thorson. The distribution of most species in this family is restricted to a single basin or fluvial system. Only Potamotrygon motoro, Potamotrygon orbignyi and Paratrygon aiereba are found in more than one river basin. In this study we investigate genetic structuring of Paratrygon aiereba, from five rivers of the Amazon region: Negro, Solimões-Amazon-Estuary system, Tapajós, Xingu and Araguaia. Sixty-three individuals were sequenced for ATPase 6, and a representative subsample of 27 individuals was sequenced for COI. The COI dataset analysis indicated that Paratrygon is sister to all other potamotrygonid genera and species. Population parameters inferred from the analysis of ATPase 6 sequences revealed that the populations of this species are structured within each river, with no or nearly non-existent gene flow occurring between rivers and a positive correlation between geographic and genetic distances. Paratrygon aiereba is comprised of three geographically restricted clades with K2P interclade distances of at least 2%. Intraspecific divergence within P. aiereba is similar to the interspecific divergence observed in Potamotrygon spp. sampled throughout the same geographic area. Using the premises of COI barcoding and the allopatric distribution of the three P. aiereba clades, the taxon P. aiereba most likely comprises three distinct biological species. Since freshwater stingrays of the family Potamotrygonidae are highly exploited for the aquarium trade, management and conservation strategies need to be implemented at the level of each river basin, rather than at the level of the Amazon basin.

A família Potamotrygonidae forma um clado monofilético com três gêneros: Paratrygon Duméril, Potamotrygon Garman e Plesiotrygon Rosa, Castello & Thorson. A maioria das espécies dessa família possui distribuição restrita a uma única bacia ou sistema fluvial, e somente as espécies Potamotrygon motoro, Potamotrygon orbignyi e Paratrygon aiereba estão presentes em mais de uma bacia hidrográfica. O presente estudo teve como objetivo investigar a estrutura genética de Paratrygon aiereba em alguns rios da região Amazônica: Negro, sistema Solimões-Amazonas, Tapajós, Xingu, e Araguaia. Para tal foram utilizados como marcador molecular os genes de ATPase subunidade 6, e COI. As análises com o fragmento de COI indicaram que o gênero Paratrygon é grupo irmão dos outros gêneros da família potamotrygonidae. Os resultados para o fragmento de ATPase mostraram que essas populações estão estruturadas dentro dos rios, com fluxo gênico restrito, ou mesmo sem fluxo gênico, apresentando uma correlação positiva entre distância genética e distância geográfica. Paratrygon aiereba é composta por três clados com distância genética de pelo menos 2%. A divergência encontrada dentro desse grupo é semelhante à observada entre Potamotrygon spp. Segundo as premissas para barcoding COI e a distribuição alopátrica de três clados em P. aiereba indicam que esse grupo pode ser um complexo de espécies. O rio Negro é conhecido por sua pesca ornamental, e na calha Solimões-Amazonas, esses animais são utilizados como fonte de proteína e sofrem com a pesca comercial. Em vista disso medidas de conservação para esta espécie devem ser tomadas em nível local, considerando cada rio separadamente, ao invés de empregar escalas regionais maiores.

Co-infection of Dengue virus by serotypes 3 and 4 in patients from Amazonas, Brazil / Co-infecção pelo vírus dengue 3 e 4 em pacientes da Amazônia brasileira

The natural co-infection with dengue virus can occur in highly endemic areas where different serotypes have been observed for many years. We report here four cases of DENV-3/DENV-4 co-infection detected by serological and molecular tests among 674 patients with acute undifferentiated fever from the tropical medicine reference center of Manaus City, Brazil, between 2005 and 2010. Analysis of the sequences obtained indicated the presence of genotype 3 and 1 for DENV-3 and DENV-4 respectively.

A co-infecção natural com os vírus dengue pode ocorre em áreas altamente endêmicas onde diferentes sorotipos têm sido transmitidos por muitos anos. Relatamos aqui quatro casos de co-infecção com DENV-3/DENV-4 detectados por testes sorológicos e moleculares entre 674 pacientes com febre indiferenciada aguda, atendidos em um centro de medicina tropical de referência da cidade de Manaus, Brasil, entre 2005 e 2010. As análises das sequências obtidas indicaram a presença dos genotipos 3 e 1 para DENV-3 e DENV-4 respectivamente.

Genomic resources for the conservation and management of the harpy eagle (Harpia harpyja, Falconiformes, Accipitridae)

We report the characterization and optimization of 45 heterologous microsatellite loci, and the development of a new set of molecular sex markers for the conservation and management of the Neotropical harpy eagle (Harpia harpyja L. 1758). Of the 45 microsatellites tested, 24 were polymorphic, six monomorphic, 10 uncharacterizable due to multiple bands and five did not amplify. The observed gene diversity of the analyzed sample of H. harpyja was low and similar to that of other threatened Falconiformes. While a high proportion of the microsatellite markers were highly variable, individuals of H. harpyja could be differentiated by a joint analysis of just three (p = 2.79 x 10-4) or four markers (p = 2.89 x 10-5). Paternity could be rejected with 95.23 percent and 97.83 percent probabilities using the same three and four markers, respectively. The sex determination markers easily and consistently differentiated males from females even with highly degraded DNA extracted from naturally shed feathers. The markers reported in this study potentially provide an excellent set of molecular tools for the conservation and management of wild and captive H. harpyja and they may also prove useful for the enigmatic Neotropical crested eagle (Morphnus guianensis Daudin 1800).

Population genetic analysis of Caiman crocodilus (Linnaeus, 1758) from South America

The genetic structure of Caiman crocodilus was investigated using a 1085 bp mtDNA fragment of the cytochrome b gene. Inferences were based on 125 individuals from nine localities in Peru, Brazil and French Guiana. With the exception of Mamirauá Lake, Anavilhanas Archipelago and the Tapará Community which show a signal of demographic expansion, the sampled localities are in a mutation-drift genetic equilibrium. Divergence between the Amazon basin and extra-Amazon basin localities is significant; however, inference from Nested Clade Analysis cannot distinguish between continuous range expansion, long distance colonization or past fragmentation; however, past fragmentation is unlikely due to low number of mutational steps separating these two regions. The divergence is probably maintained by the reduced ability of C. crocodilus to cross salt water barriers. Within the Amazon basin, continuous range expansion without isolation-by-distance is the most likely process causing genetic structuring. The observed genetic patterns are compatible with the ecology of C. crocodilus, and history of human exploitation. As commercial hunting depleted more valuable species, C. crocodilus expanded its range and ecological niche, prompting hunters to harvest it. Following a period of intense hunting, C. crocodilus is now experiencing recovery and a second population expansion especially in protected areas.