Chromosomal evolution of neotropical cichlids: The role of repetitive DNA sequences in the organization and structure of karyotype

Cichlids are important in the aquaculture and ornamental fish trade and are considered models for evolutionary biology. However, most studies of cichlids have investigated African species, and the South American cichlids remain poorly characterized. Studies in neotropical regions have focused almost exclusively on classical cytogenetic approaches without investigating physical chromosomal mapping of specific sequences. The aim of the present study is to investigate the genomic organization of species belonging to different tribes of the subfamily Cichlinae (Cichla monoculus, Astronotus ocellatus, Geophagus proximus, Acaronia nassa, Bujurquina peregrinabunda, Hoplarchus psittacus, Hypselecara coryphaenoides, Hypselecara temporalis, Caquetaia spectabilis, Uaru amphiacanthoides, Pterophyllum leopoldi, Pterophyllum scalare, and Symphysodon discus) and reexamine the karyotypic evolutionary patterns proposed for this group. Variations in some cytogenetic markers were observed, although no trends were found in terms of the increase, decrease, or maintenance of the basal diploid chromosome number 2n = 48 in the tribes. Several species were observed to have 18S rDNA genetic duplications, as well as multiple rDNA loci. In most of the taxa analyzed, the 5S rDNA was located in the interstitial region of a pair of homologous chromosomes, although variations from this pattern were observed. Interstitial telomere sites were also observed and appear to be involved in chromosomal rearrangement events and the accumulation of repeat-rich satellite DNA sequences. Our data demonstrated the karyotypic diversity that exists among neotropical cichlids, suggesting that most of this diversity is due to the repetitive sequences present in heterochromatic regions and that repeat sequences have greatly influenced the karyotypic evolution of these fishes. © 2012 Springer Science+Business Media B.V.

Composição florística de angiospermas no carste do Alto São Francisco, Minas Gerais, Brasil

The occurence of carbonatic rocks defines a typical type of relief, called karst, that, by its geomorphology and hydrology, usually differs from the surrounding landscape. In the upper São Francisco river basin, some striking remnants of vegetation associated to karst can be found, which are locally knwon as Mata de Pains. In this region, a mosaic of different physiognomies, including forests and open areas, which present noteworthy plant diversity, composes the vegetation. The aim of this study is to provide an inventory of angiosperm species in areas of carbonatic rocks outcrops in the upper São Francisco river basin karst region, as well as analyze the floristic relationship of the study area with different Brazilian phytogeographic domains. Fieldwork was performed during the period of 2002 a 2006, when collections of fertile speciemns were done in areas associated to carbonatic rock outcrops. During the study, 1512 exsicates were incorporated to BHCB herbarium collection. A total of 456 angiosperm species were inventoried, distributed in 299 genera and 77 families. Herbaceous habit was better represented, with 161 species, followed by shrubs and trees (111 species each) and lianas (73 species). The flora of the study area presents more influence of the Mata Atlântica domain (Atlantic Rain Forest), followed by, in order of importance, Cerrado, Amazonia, Caatinga, Pantanal and Pampa. Floristic surveys in different karst regions inserted in different phytogeographic domains may, together, provide useful information in understanding the phytogeografic history of Neotropical vegetation.

Development of microsatellite markers for the Neotropical endemic Brazilian Guanabara frog, Euparkerella brasiliensis, through 454 shotgun pyrosequencing

The new-generation 454 GS-FLX Titanium pyrosequencing was used to isolate microsatellite markers for the Brazilian Guanabara frog, Euparkerella brasiliensis, an Atlantic forest endemic species. Three multiplex polymerase chain reaction sets were optimized for genotyping of 11 polymorphic (di- and tetranucleotide) microsatellite markers. Genetic diversity was assessed in 21 individuals from a population (Reserva Ecológica de Guapiaçu, REGUA) locatedin the central region of the Rio de Janeiro State, in Brazil. The mean number of alleles per locus ranged from 3 to 12. Observed and expected heterozygosities ranged from 0.095 to 0.905 and from 0.094 to 0.904, respectively. After using the Bonferroni correction for multiple tests, there was no evidence of linkage disequilibrium between pairs of loci but deviations for Hardy-Weinberg equilibrium were found in 4 loci. We found no evidence for allele dropouts or stuttering, but we detected the presence of null alleles at loci Eb10 and Eb36. These markers will be useful for analyses of fine-scale population structure and determination of relative effects of habitat loss and fragmentation on population genetic variability within species. © FUNPEC-RP.

Two new species of Diaptomidae (Crustacea: Copepoda: Calanoida) from the Neotropical Region (Paraná River)

Two new species of Diaptomidae were found in the Middle Paraná River. The new species are referred to as Diaptomus curvatus sp. nov. and Diaptomus frutosae sp. nov. Important character states in Diaptomus curvatus include: right fifth leg with a well-developed endopod and with the lateral spine of the second exopod segment curved and longer than the segment on which it is inserted, plus the presence of a large dorsal process on urosomite 4 of the male. For Diaptomus frutosae they include: a differentiated hook-like process on segment 20 with chitinous protuberances at base of male right antennule; the last segment of the right leg 5 of male is triangular, and the lateral spine inserted distal to mid-level of this segment. Both species are placed in Diaptomus sensu lato as a temporary placement. http://www.zoobank.org/urn:lsid:zoobank.org:pub:FECDD6C4-C0AC-4043-ADDD-F29539B451F0. © 2013 Copyright Taylor and Francis Group, LLC.

Can DNA barcoding accurately discriminate megadiverse Neotropical freshwater fish fauna?

Background: The megadiverse Neotropical freshwater ichthyofauna is the richest in the world with approximately 6,000 recognized species. Interestingly, they are distributed among only 17 orders, and almost 80% of them belong to only three orders: Characiformes, Siluriformes and Perciformes. Moreover, evidence based on molecular data has shown that most of the diversification of the Neotropical ichthyofauna occurred recently. These characteristics make the taxonomy and identification of this fauna a great challenge, even when using molecular approaches. In this context, the present study aimed to test the effectiveness of the barcoding methodology (COI gene) to identify the mega diverse freshwater fish fauna from the Neotropical region. For this purpose, 254 species of fishes were analyzed from the Upper Parana River basin, an area representative of the larger Neotropical region.Results: Of the 254 species analyzed, 252 were correctly identified by their barcode sequences (99.2%). The main K2P intra- and inter-specific genetic divergence values (0.3% and 6.8%, respectively) were relatively low compared with similar values reported in the literature, reflecting the higher number of closely related species belonging to a few higher taxa and their recent radiation. Moreover, for 84 pairs of species that showed low levels of genetic divergence (<2%), application of a complementary character-based nucleotide diagnostic approach proved useful in discriminating them. Additionally, 14 species displayed high intra-specific genetic divergence (>2%), pointing to at least 23 strong candidates for new species.Conclusions: Our study is the first to examine a large number of freshwater fish species from the Neotropical area, including a large number of closely related species. The results confirmed the efficacy of the barcoding methodology to identify a recently radiated, megadiverse fauna, discriminating 99.2% of the analyzed species. The power of the barcode sequences to identify species, even with low interspecific divergence, gives us an idea of the distribution of inter-specific genetic divergence in these megadiverse fauna. The results also revealed hidden genetic divergences suggestive of reproductive isolation and putative cryptic speciation in some species (23 candidates for new species). Finally, our study constituted an important contribution to the international Barcoding of Life (iBOL.org) project, providing barcode sequences for use in identification of these species by experts and non-experts, and allowing them to be available for use in other applications. © 2013 Pereira et al.; licensee BioMed Central Ltd.

Water availability determines the richness and density of fig trees within Brazilian semideciduous forest landscapes

The success of fig trees in tropical ecosystems is evidenced by the great diversity (+750 species) and wide geographic distribution of the genus. We assessed the contribution of environmental variables on the species richness and density of fig trees in fragments of seasonal semideciduous forest (SSF) in Brazil. We assessed 20 forest fragments in three regions in Sao Paulo State, Brazil. Fig tree richness and density was estimated in rectangular plots, comprising 31.4 ha sampled. Both richness and fig tree density were linearly modeled as function of variables representing (1) fragment metrics, (2) forest structure, and (3) landscape metrics expressing water drainage in the fragments. Model selection was performed by comparing the AIC values (Akaike Information Criterion) and the relative weight of each model (wAIC). Both species richness and fig tree density were better explained by the water availability in the fragment (meter of streams/ha): wAICrichness = 0.45, wAICdensity = 0.96. The remaining variables related to anthropic perturbation and forest structure were of little weight in the models. The rainfall seasonality in SSF seems to select for both establishment strategies and morphological adaptations in the hemiepiphytic fig tree species. In the studied SSF, hemiepiphytes established at lower heights in their host trees than reported for fig trees in evergreen rainforests. Some hemiepiphytic fig species evolved superficial roots extending up to 100 m from their trunks, resulting in hectare-scale root zones that allow them to efficiently forage water and soil nutrients. The community of fig trees was robust to variation in forest structure and conservation level of SSF fragments, making this group of plants an important element for the functioning of seasonal tropical forests. © 2013 Elsevier Masson SAS. All rights reserved.

Structure, histochemistry and ultrastructure of the male reproductive accessory glands in the neotropical flat-faced fruit-eating bat Artibeus planirostris (Chiroptera: Phyllostomidae)

Chiroptera, the second largest mammalian order, presents different reproductive strategies and unique reproductive features. However, there are few reports regarding male reproductive accessory glands (RAGs) in Chiroptera. Thus, the aim of the present study was to characterise the RAGs of the exclusively neotropical bat Artibeus planirostris (Chiroptera: Phyllostomidae) macroscopically, microscopically and ultrastructurally. The RAGs were composed of a prostatic complex with two regions (ventral and dorsal) and paraurethral and bulbourethral glands, but no seminal vesicles. The ventral region had an undefined epithelium, with secretory and basal cells, and its secretions were periodic acid-Schiff (PAS) positive. The dorsal region received both deferens ducts, had a columnar pseudostratified epithelium with secretory and basal cells. There were two types of secretions from the dorsal region: one that was basophilic and another that was mixed PAS positive and PAS negative. The paraurethral glands were dispersed in the connective tissue of the urethra, whereas the bulbourethral glands were located in the penile root. Histological and ultrastructural data confirmed the prostatic nature of the ventral and dorsal regions and the holocrine nature of the ventral region, with the latter finding never having been described previously for the prostate gland. Our findings demonstrate the wide discrepancy of RAGs between A. planirostris and other mammals in terms of their composition, structure and morphology. © CSIRO 2013.

Morphological adaptations to arboreal habitats and heart position in species of the neotropical whipsnakes genus Chironius

The evolution of arboreality in snakes is accompanied by modifications that are remarkably similar across species. Gravity is one of the most important selective agents, and arboreal snakes present adaptations to circumvent the gradient of pressure, including modifications on heart position (HP) and body slenderness (BS). However, the degree to which different life-history traits influence the cardiovascular system of snakes remains unclear. Here, we used an ecological and a phylogenetic approach to explore the relationship between habitat, HP, BS, and heart size (HS) in five species of the neotropical whipsnakes genus Chironius that occupy terrestrial, semiarboreal, and arboreal habits. Our ecological comparison indicated that the arboreal species have the most posterior-positioned heart, the most slender body, and the smallest HS, whereas the terrestrial representative of the group exhibited the most anterior heart, the less flattened body, and the largest HS. After removing the phylogenetic effect, we found no difference in HP and BS between terrestrial and arboreal species. Habitat only differed when contrasting with HS. Body slenderness and HS were correlated with HP. Our results suggest that different restrictions, such as anatomical constraints, behavior, and phylogenetic inertia, may be important for the studied species. © 2013 The Royal Swedish Academy of Sciences.

New Neotropical species of Hygronemobius Hebard, 1913 (Orthoptera: Grylloidea: Nemobiinae), including a brief discussion of male genitalia morphology and preliminary biogeographic considerations of the genus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The pioneering use of ISSR (Inter Simple Sequence Repeat) in Neotropical anurans: Preliminary assessment of genetic diversity in populations of Physalaemus cuvieri (Amphibia, Leiuperidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Mixing Eucalyptus and Acacia trees leads to fine root over-yielding and vertical segregation between species

The consequences of diversity on belowground processes are still poorly known in tropical forests. The distributions of very fine roots (diameter <1 mm) and fine roots (diameter <3 mm) were studied in a randomized block design close to the harvest age of fast-growing plantations. A replacement series was set up in Brazil with mono-specific Eucalyptus grandis (100E) and Acacia mangium (100A) stands and a mixture with the same stocking density and 50 % of each species (50A:50E). The total fine root (FR) biomass down to a depth of 2 m was about 27 % higher in 50A:50E than in 100A and 100E. Fine root over-yielding in 50A:50E resulted from a 72 % rise in E. grandis fine root biomass per tree relative to 100E, whereas A. mangium FR biomass per tree was 17 % lower than in 100A. Mixing A. mangium with E. grandis trees led to a drop in A. mangium FR biomass in the upper 50 cm of soil relative to 100A, partially balanced by a rise in deep soil layers. Our results highlight similarities in the effects of directional resources on leaf and FR distributions in the mixture, with A. mangium leaves below the E. grandis canopy and a low density of A. mangium fine roots in the resource-rich soil layers relative to monospecific stands. The vertical segregation of resource-absorbing organs did not lead to niche complementarity expected to increase the total biomass production. © 2012 Springer-Verlag Berlin Heidelberg.

Composition and interrelationships of a large Neotropical freshwater fish group, the subfamily Cheirodontinae (Characiformes: Characidae): A case study based on mitochondrial and nuclear DNA sequences

•Relationships of Cheirodontinae based on a broad taxonomic sample.•Results reject the monophyly of Cheirodontinae as previously conceived.•Exclusion of Amazonspinther and Spintherobolus from the subfamily Cheirodontinae.•The removal of Leptagoniates pi of the genus Leptagoniates and inclusion in Cheirodontinae.•Division of Cheirodontinae in three newly defined monophyletic tribes. Characidae is the most species-rich family of freshwater fishes in the order Characiformes, with more than 1000 valid species that correspond to approximately 55% of the order. Few hypotheses about the composition and internal relationships within this family are available and most fail to reach an agreement. Among Characidae, Cheirodontinae is an emblematic group that includes 18 genera (1 fossil) and approximately 60 described species distributed throughout the Neotropical region. The taxonomic and systematic history of Cheirodontinae is complex, and only two hypotheses about the internal relationships in this subfamily have been reported to date. In the present study, we test the composition and relationships of fishes assigned to Cheirodontinae based on a broad taxonomic sample that also includes some characid incertae sedis taxa that were previously considered to be part of Cheirodontinae. We present phylogenetic analyses of a large molecular dataset of mitochondrial and nuclear DNA sequences. Our results reject the monophyly of Cheirodontinae as previously conceived, as well as the tribes Cheirodontini and Compsurini, and the genera Cheirodon, Compsura, Leptagoniates, Macropsobrycon, Odontostilbe, and Serrapinnus. On the basis of these results we propose: (1) the exclusion of Amazonspinther and Spintherobolus from the subfamily Cheirodontinae since they are the sister-group of all remaining Characidae; (2) the removal of Macropsobrycon xinguensis of the genus Macropsobrycon; (3) the removal of Leptagoniates pi of the genus Leptagoniates; (4) the inclusion of Leptagoniates pi in the subfamily Cheirodontinae; (5) the removal of Cheirodon stenodon of the genus Cheirodon and its inclusion in the subfamily Cheirodontinae under a new genus name; (6) the need to revise the polyphyletic genera Compsura, Odontostilbe, and Serrapinnus; and (7) the division of Cheirodontinae in three newly defined monophyletic tribes: Cheirodontini, Compsurini, and Pseudocheirodontini. Our results suggest that our knowledge about the largest Neotropical fish family, Characidae, still is incipient. © 2013 Elsevier Inc..

Phylogeographic Structure And Outbreeding Depression Reveal Early Stages Of Reproductive Isolation In The Neotropical Orchid Epidendrum Denticulatum

Phylogeographic studies provide an important framework for investigating the mechanisms operating during the earliest stages of speciation, as reproductive barriers can be examined among divergent lineages in a geographic context. We investigated the evolution of early stages of intrinsic postmating isolation among different populations and lineages of Epidendrum denticulatum, a Neotropical orchid distributed across different biomes in South America. We estimated genetic diversity and structure for both nuclear and plastid markers, using a haplotype network, differentiation tests, Bayesian assignment analysis, and divergence time estimates of the main lineages. Reproductive barriers among divergent lineages were examined by analyzing seed viability following reciprocal crossing experiments. Strong plastid phylogeographic structure was found, indicating that E. denticulatum was restricted to multiple refuges during South American forest expansion events. In contrast, significant phylogeographic structure was not found for nuclear markers, suggesting higher gene flow by pollen than by seeds. Large asymmetries in seed set were observed among different plastid genetic groups, suggesting the presence of polymorphic genic incompatibilities associated with cytonuclear interactions. Our results confirm the importance of phylogeographic studies associated with reproductive isolation experiments and suggest an important role for outbreeding depression during the early stages of lineage diversification. © 2013 The Society for the Study of Evolution.

Applying machine learning based on multiscale classifiers to detect remote phenology patterns in Cerrado savanna trees

Plant phenology is one of the most reliable indicators of species responses to global climate change, motivating the development of new technologies for phenological monitoring. Digital cameras or near remote systems have been efficiently applied as multi-channel imaging sensors, where leaf color information is extracted from the RGB (Red, Green, and Blue) color channels, and the changes in green levels are used to infer leafing patterns of plant species. In this scenario, texture information is a great ally for image analysis that has been little used in phenology studies. We monitored leaf-changing patterns of Cerrado savanna vegetation by taking daily digital images. We extract RGB channels from the digital images and correlate them with phenological changes. Additionally, we benefit from the inclusion of textural metrics for quantifying spatial heterogeneity. Our first goals are: (1) to test if color change information is able to characterize the phenological pattern of a group of species; (2) to test if the temporal variation in image texture is useful to distinguish plant species; and (3) to test if individuals from the same species may be automatically identified using digital images. In this paper, we present a machine learning approach based on multiscale classifiers to detect phenological patterns in the digital images. Our results indicate that: (1) extreme hours (morning and afternoon) are the best for identifying plant species; (2) different plant species present a different behavior with respect to the color change information; and (3) texture variation along temporal images is promising information for capturing phenological patterns. Based on those results, we suggest that individuals from the same species and functional group might be identified using digital images, and introduce a new tool to help phenology experts in the identification of new individuals from the same species in the image and their location on the ground. © 2013 Elsevier B.V. All rights reserved.

Conjunctival flora, Schirmer's tear test, intraocular pressure, and conjunctival cytology in neotropical primates from Salvador, Brazil

Background: This study aimed to establish reference values for selected ophthalmic diagnostic tests in healthy neotropical primates from Salvador, Brazil. Methods: A total of 73 intact adults, including Callithrix jacchus (n = 31), Callithrix penicillata (n = 8), Cebus sp. (n = 22), and Cebus xanthosternos (n = 9) were used to evaluate the normal conjunctival bacterial flora. Cebus xanthosternos (n = 12) were used to evaluate tear production with Schirmer's tear test (STT), intraocular pressure (IOP), and conjunctival cytology. Results: For all animals evaluated, Gram-positive bacteria were predominant. Results of the diagnostic tests in Cebus xanthosternos were as follows: STT: 14.92 ± 5.46 mm/minutes, IOP: 19.62 ± 4.57 mmHg, and conjunctival cytology revealed intermediate squamous epithelial cells in great quantities. Conclusions: These ophthalmic reference values will be particularly useful to diagnose discrete or unusual pathological changes in the neotropical primates eye. © 2013 John Wiley & Sons A/S.