Hybridization and massive mtDNA unidirectional introgression between the closely related Neotropical toads Rhinella marina and R. schneideriinferred from mtDNA and nuclear markers

BACKGROUND: The classical perspective that interspecific hybridization in animals is rare has been changing due to a growing list of empirical examples showing the occurrence of gene flow between closely related species. Using sequence data from cyt b mitochondrial gene and three intron nuclear genes (RPL9, c-myc, and RPL3) we investigated patterns of nucleotide polymorphism and divergence between two closely related toad species R. marina and R. schneideri. By comparing levels of differentiation at nuclear and mtDNA levels we were able to describe patterns of introgression and infer the history of hybridization between these species. RESULTS: All nuclear loci are essentially concordant in revealing two well differentiated groups of haplotypes, corresponding to the morphologically-defined species R. marina and R. schneideri. Mitochondrial DNA analysis also revealed two well-differentiated groups of haplotypes but, in stark contrast with the nuclear genealogies, all R. schneideri sequences are clustered with sequences of R. marina from the right Amazon bank (RAB), while R. marina sequences from the left Amazon bank (LAB) are monophyletic. An Isolation-with-Migration (IM) analysis using nuclear data showed that R. marina and R. schneideri diverged at ≈ 1.69 Myr (early Pleistocene), while R. marina populations from LAB and RAB diverged at ≈ 0.33 Myr (middle Pleistocene). This time of divergence is not consistent with the split between LAB and RAB populations obtained with mtDNA data (≈ 1.59 Myr), which is notably similar to the estimate obtained with nuclear genes between R. marina and R. schneideri. Coalescent simulations of mtDNA phylogeny under the speciation history inferred from nuclear genes rejected the hypothesis of incomplete lineage sorting to explain the conflicting signal between mtDNA and nuclear-based phylogenies. CONCLUSIONS: The cytonuclear discordance seems to reflect the occurrence of interspecific hybridization between these two closely related toad species. Overall, our results suggest a phenomenon of extensive mtDNA unidirectional introgression from the previously occurring R. schneideri into the invading R. marina. We hypothesize that climatic-induced range shifts during the Pleistocene/Holocene may have played an important role in the observed patterns of introgression

PRIMEIRO REGISTRO DE Cymothoa sp. FABRICIUS, 1793 (ISOPODA: CYMOTHOIDAE) PARASITANDO O PEIXE DE QUATRO OLHOS, Anableps anableps (CYPRINODONTIFORMES: ANABLEPIDAE), NA COSTA NORTE BRASILEIRA

This study presents the first record of the occurrence of Cymothoa Fabricius, 1793 in northern Brazil, as well as in four-eyed fish Anableps anableps. The research was conducted in the Curuçá estuary, in northern Brazil, where 158 specimens of A. anableps were collected, with nine Cymothoa specimens being found on nine different fish. These isopods were identified by sequencing the mitochondrial Cytochrome Oxidase I gene. The prevalence of the parasite (4.43 %) in A. anableps was extremely low and Cymothoa sp. specimens were found primarily in the gills of the host fish.Keywords: Amazon; Isopoda; Crustaceans; Four-Eyed Fish; Anablepidae.Este estudo apresenta o primeiro registro da ocorrência de Cymothoa Fabricius, 1793 no norte do Brasil, bem como no peixe de quatro olhos Anableps anableps. A pesquisa foi realizada no estuário de Curuçá, no norte do Brasil, onde foram coletados 158 espécimes de A. anableps, com nove espécimes de Cymothoa, sendo encontrados em nove peixes diferentes. Esses isópodes foram identificados pelo sequenciamento do gene mitocondrial do citocromo oxidase I. A prevalência do parasita (4.43 %) em A. anableps foi extremamente baixa e os espécimes de Cymothoa foram encontrados principalmente nas brânquias do peixe hospedeiro.Palavras-chave: Amazônia, Isopoda, Crustáceos, Peixe com quatro olhos, Anablepidae

Mitochondrial DNA-like sequence in the nuclear genome of Saguinus (Callitrichinae, Primates): transfer estimation

Mitochondrial DNA-like sequences have been found in the nuclei of a variety of organisms. These nuclear pseudogenes can be used to estimate relative evolutionary rates of mitochondrial genes, and can be used as outgroups in phylogenetic analyses. In this study, mitochondrial sequences with pseudogene-like characteristics, including deletions and/or insertions and stop codons, were found in tamarins (Saguinus spp., Callitrichinae, Primates). Phylogenetic analysis allowed estimation of the timing of the migration of these sequences to the nuclear genome, and also permitted inferences on the phylogeny of the genus. The choice of an inadequate outgroup (Aotus infulatus) prevented a good phylogenetic resolution of the subfamily Callitrichinae. The relatively ancient divergence of the Cebidae (Callitrichinae, Aotinae and Cebinae) may have favored confounding homoplasies.Seqüências tipo mitocondriais têm comumente sido encontradas no genoma nuclear de diversos organismos. Quando acidentalmente incluídas em estudos de seqüências mitocondriais, diversas conclusões errôneas podem ser obtidas. No entanto, estes pseudogenes nucleares tipo mitocondriais podem ser usados para a estimativa da taxa relativa de evolução de genes mitocondriais e também como grupo externo em análises filogenéticas. No presente trabalho, seqüências mitocondriais com características do tipo de pseudogene, tais como deleções e/ou inserções e códons de parada, foram encontradas em tamarins (Saguinus spp., Callitrichinae, Primates). A análise filogenética permitiu a estimativa do tempo da migração da seqüência mitocondrial para o genoma nuclear e algumas inferências filogenéticas. A escolha de um grupo externo não adequado (Aotus infulatus) não permitiu uma reconstrução filogenética confiável da subfamília Callitrichinae. A divergência bastante antiga de Cebidae (Callitrichinae, Aotinae e Cebinae) pode ter favorecido o aparecimento de homoplasias, obscurecendo a análise

Genetic characterisation of populations of the critically endangered Goliath grouper (Epinephelus itajara, Serranidae) from the Northern Brazilian coast through analyses of mtDNA

The Goliath grouper (Epinephelus itajara) is one of the most endangered species of fish of the subfamily Epinephelinae. Slow to develop and mature, and dependent on mangrove habitats for breeding, the species also suffers intense harvesting, which has reduced drastically in numbers in many areas. To contribute to the understanding of the characteristics of E. itajara populations, we conducted a molecular genetics study of the species, focusing on populations from the Northern Brazilian coast. The mtDNA control region (D-loop) of 116 individuals from five localities (Bragança, Ajuruteua, Parnaíba, Fortaleza and Natal) was analysed, and a sequence of 499 base pairs identified. Analyses of the sequences indicated that genetic variability was generally lower in E. itajara than in other endangered species of the genus. AMOVA found no significant grouping structure among the populations. Nested Clade Analysis revealed a significant association between genetic variability and geographic distribution among only three populations (Ajuruteua, Parnaíba and Natal). Genetic diversity was higher in populations from the Amazon region, which may be related to the better conservation of mangrove habitats in this area. Therefore, the present study could be used for the implementation of conservation and management measures in order to protect and consolidate these populations

Molecular phylogeny of the genus Saguinus (Platyrrhini, Primates) based on the ND1 mitochondrial gene and implications for conservation

The systematics of the subfamily Callitrichinae (Platyrrhini, Primates), a group of small monkeys from South America and Panama, remains an area of considerable discussion despite many investigations, there being continuing controversy over subgeneric taxonomic classifications based on morphological characters. The purpose of our research was to help elucidate the phylogenetic relationships within the monkey genus Saguinus (Callitrichinae) using a molecular approach to discover whether or not the two different sections containing hairy-faced and bare-faced species are monophyletic, whether Saguinus midas midas and Saguinus bicolor are more closely related than are S. midas midas and Saguinus midas niger, and if Saguinus fuscicollis melanoleucus and Saguinus fuscicollis weddelli really are different species. We sequenced the 957 bp ND1 mitochondrial gene of 21 Saguinus monkeys (belonging to six species and nine morphotypes) and one Cebus monkey (the outgroup) and constructed phylogenetic trees using maximum parsimony, neighbor joining, and maximum likelihood methods. The phylogenetic trees obtained divided the genus Saguinus into two groups, one containing the small-bodied species S. fuscicollis and the other, the large-bodied species S. mystax, S. leucopus, S. oedipus, S. midas, S. bicolor. The most derived taxa, S. midas and S. bicolor, grouped together, while S. fuscicollis melanoleucus and S. f. weddelli showed divergence values that did not support the division of these morphotypes into subspecies. On the other hand, S. midas individuals showed divergence compatible with the existence of three subspecies, two of them with the same morphotype as the subspecies S. midas niger. The results of our study suggest that there is at least one Saguinus subspecies that has not yet been described and that the conservation status of Saguinus species and subspecies should be carefully revised using modern molecular approaches

Commercialization of a critically endangered species (largetooth sawfish, Pristis perotteti) in fish markets of northern Brazil: Authenticity by DNA analysis

AbstractPristis perotteti, the largetooth sawfish, is one of most endangered elasmobranchs because of fisheries and habitat degradation. Its commercialization in Brazil is prohibited, but fresh or salted fillets of this fish can be found in markets, labeled as “sharks”. In this study we performed genetic analyses on “shark” samples from two important fishery-trading ports in northern Brazil (Vigia and Bragança). Based on partial DNA sequences of the mitochondrial 16S and Cyt b genes, 24 (55%) out of 44 samples were unequivocally identified as P. perotteti while the others comprised eight species of the families Carcharhinidae and Ginglymostomatidae. These results show that fishing surveillance and monitoring have not been effective to prohibit the commercialization of this highly endangered species

Low levels of genetic diversity depicted from mitochondrial DNA sequences in a heavily exploited marine fish (Cynoscion acoupa, Sciaenidae) from the Northern coast of Brazil

The acoupa weakfish (Cynoscion acoupa - Sciaenidae) is a marine species of croaker with estuarine-dependent behavior, found in the western Atlantic from Panama to Argentina. It is one of the most exploited food fish on the northern coast of Brazil. In this study, DNA sequences were determined from the entire control region (D-loop) of the mitochondrial genome of 297 individuals collected during seven different months between December 2003 and August 2005 on the northern coast of Brazil (Amapá and Pará). Genetic variability expressed by haplotype (h = 0,892) and nucleotide (p = 0,003) diversities were low compared to other heavily exploited marine fish species from the western Atlantic and eastern Asia. AMOVA depicted a lack of genetic structuring among the samples from different years, indicating the presence of a single stock of C. acoupa within the sample area. The possible reasons for the low levels of genetic diversity are discussed. These results demonstrate a need for the monitoring of C. acoupa harvesting and the preservation of the estuaries within its geographic range, considering that this large fish depends on estuarine ecosystems during part of its life cycle

Análise populacional de Scomberomorus cavalla (Cuvier, 1829) (Perciformes, Scombridae) da costa Norte e Nordeste brasileira

Scomberomorus cavalla is a pelagic fish species widely distributed on the Atlantic west coast, and a noticeable decrease in its capture level in the USA and Gulf of Mexico is occurring, compared to the levels reached by the species in the past. Likewise, in some areas of Brazil, there has been indication of over-harvesting. However, there are no molecular studies focusing on the management of such an important item. Thus, in the present study, 380 nucleotide base pairs of the mitochondrial DNA D-Loop region of samples from Macapá, Bragança, and Fortaleza were sequenced. Phylogenetic and population analyses revealed that there is only one panmitic population, and low levels of genetic variability were verified. These results, as well as the noticed over-harvesting of S. cavalla, represent very important data to determine the management of such stock in order to prevent a collapse or the risk of future extinction.Scomberomorus cavalla é uma espécie de peixe pelágico amplamente distribuído na costa oeste do Atlântico, e uma diminuição no seu nível de captura tem sido verificada nos E.U.A e Golfo do México, comparada com os níveis alcançados pela espécie no passado. Da mesma forma, em algumas áreas do Brasil, há indícios de sobre-exploração. Entretanto, não existem estudos moleculares que visam o manejo deste importante item. Desta forma, no presente estudo, foram seqüenciados 380 pares de bases nucleotídicas da região da Alça-D do DNA mitocondrial de amostras provenientes de desembarque em Macapá, Bragança e Fortaleza. As análises filogenéticas e populacionais revelaram que há apenas uma população panmítica e baixos níveis de variabilidade genética foram observados. Estes resultados, assim como a observada sobre-exploração de S. cavala, representam dados muito importantes para o estabelecimento do manejo deste estoque a fim de prevenir um colapso ou risco de extinção no futuro