Food web structure of fish communities of Doce River, 5 years after the Fundão dam failure.

The rupture of the Fundão dam is considered the largest mining failure in history, which had a particularly detrimental impact on fish populations, as the mud from the ore tailings significantly altered the water quality and habitat of Doce River basin. This study aimed to assess the trophic structure of fish communities in areas impacted and not impacted by the dam rupture in the Doce River basin. To evaluate the food web structure, community-wide trophic niche, and trophic positions of fish, stable isotopes of carbon (δ13C) and nitrogen (δ15N) were utilized across ten sites (seven impacted and three control). In general, fish appeared to assimilate resources such as invertebrates, algae, and periphyton, although the importance of each resource varied among sites. The site closest to the dam rupture exhibited a more simplified trophic structure compared to the control sites and those nearer the river mouth. In this site, most fish species occupied a similar trophic position. Trophic niches also exhibited the greatest dissimilarity between the site closest to the dam failure and those farther away from it, with an expansion of trophic niche breadth observed with an increase in the distance from the dam rupture. Our study provided valuable insights into the trophic structure of fish communities within the Doce River basin, shedding light on the trophic ecology of the 59 fish species investigated. We also emphasize the importance of our study for future assessments of ore tailings dam failure disasters and evaluating the effectiveness of mitigation measures for Doce River basin recovery.

The gut microbiome of exudivorous marmosets in the wild and captivity.

Mammalian captive dietary specialists like folivores are prone to gastrointestinal distress and primate dietary specialists suffer the greatest gut microbiome diversity losses in captivity compared to the wild. Marmosets represent another group of dietary specialists, exudivores that eat plant exudates, but whose microbiome remains relatively less studied. The common occurrence of gastrointestinal distress in captive marmosets prompted us to study the Callithrix gut microbiome composition and predictive function through bacterial 16S ribosomal RNA V4 region sequencing. We sampled 59 wild and captive Callithrix across four species and their hybrids. Host environment had a stronger effect on the gut microbiome than host taxon. Wild Callithrix gut microbiomes were enriched for Bifidobacterium, which process host-indigestible carbohydrates. Captive marmoset guts were enriched for Enterobacteriaceae, a family containing pathogenic bacteria. While gut microbiome function was similar across marmosets, Enterobacteriaceae seem to carry out most functional activities in captive host guts. More diverse bacterial taxa seem to perform gut functions in wild marmosets, with Bifidobacterium being important for carbohydrate metabolism. Captive marmosets showed gut microbiome composition aspects seen in human gastrointestinal diseases. Thus, captivity may perturb the exudivore gut microbiome, which raises implications for captive exudivore welfare and calls for husbandry modifications.

Genomic skimming and nanopore sequencing uncover cryptic hybridization in one of world's most threatened primates.

The Brazilian buffy-tufted-ear marmoset (Callithrix aurita), one of the world's most endangered primates, is threatened by anthropogenic hybridization with exotic, invasive marmoset species. As there are few genetic data available for C. aurita, we developed a PCR-free protocol with minimal technical requirements to rapidly generate genomic data with genomic skimming and portable nanopore sequencing. With this direct DNA sequencing approach, we successfully determined the complete mitogenome of a marmoset that we initially identified as C. aurita. The obtained nanopore-assembled sequence was highly concordant with a Sanger sequenced version of the same mitogenome. Phylogenetic analyses unexpectedly revealed that our specimen was a cryptic hybrid, with a C. aurita phenotype and C. penicillata mitogenome lineage. We also used publicly available mitogenome data to determine diversity estimates for C. aurita and three other marmoset species. Mitogenomics holds great potential to address deficiencies in genomic data for endangered, non-model species such as C. aurita. However, we discuss why mitogenomic approaches should be used in conjunction with other data for marmoset species identification. Finally, we discuss the utility and implications of our results and genomic skimming/nanopore approach for conservation and evolutionary studies of C. aurita and other marmosets.

Mitogenomic phylogeny of Callithrix with special focus on human transferred taxa.

BACKGROUND: Callithrix marmosets are a relatively young primate radiation, whose phylogeny is not yet fully resolved. These primates are naturally para- and allopatric, but three species with highly invasive potential have been introduced into the southeastern Brazilian Atlantic Forest by the pet trade. There, these species hybridize with each other and endangered, native congeners. We aimed here to reconstruct a robust Callithrix phylogeny and divergence time estimates, and identify the biogeographic origins of autochthonous and allochthonous Callithrix mitogenome lineages. We sequenced 49 mitogenomes from four species (C. aurita, C. geoffroyi, C. jacchus, C. penicillata) and anthropogenic hybrids (C. aurita x Callithrix sp., C. penicillata x C. jacchus, Callithrix sp. x Callithrix sp., C. penicillata x C. geoffroyi) via Sanger and whole genome sequencing. We combined these data with previously published Callithrix mitogenomes to analyze five Callithrix species in total. RESULTS: We report the complete sequence and organization of the C. aurita mitogenome. Phylogenetic analyses showed that C. aurita was the first to diverge within Callithrix 3.54 million years ago (Ma), while C. jacchus and C. penicillata lineages diverged most recently 0.5 Ma as sister clades. MtDNA clades of C. aurita, C. geoffroyi, and C. penicillata show intraspecific geographic structure, but C. penicillata clades appear polyphyletic. Hybrids, which were identified by phenotype, possessed mainly C. penicillata or C. jacchus mtDNA haplotypes. The biogeographic origins of mtDNA haplotypes from hybrid and allochthonous Callithrix were broadly distributed across natural Callithrix ranges. Our phylogenetic results also evidence introgression of C. jacchus mtDNA into C. aurita. CONCLUSION: Our robust Callithrix mitogenome phylogeny shows C. aurita lineages as basal and C. jacchus lineages among the most recent within Callithrix. We provide the first evidence that parental mtDNA lineages of anthropogenic hybrid and allochthonous marmosets are broadly distributed inside and outside of the Atlantic Forest. We also show evidence of cryptic hybridization between allochthonous Callithrix and autochthonous C. aurita. Our results encouragingly show that further development of genomic resources will allow to more clearly elucidate Callithrix evolutionary relationships and understand the dynamics of Callithrix anthropogenic introductions into the Brazilian Atlantic Forest.

Cytogenetics of Four Species of the Green Clade Aplastodiscus Lutz, 1950 (Anura: Cophomantinae): New Insights into the Chromosomal Evolution of the Genus.

The tree frog Aplastodiscus is a Neotropical taxon that encompasses 15 species in the Atlantic forest biome, with one isolated species in the Central Brazilian Cerrado. To date, only 8 species have been karyotyped, showing high levels of diploid number variation, which allowed clustering species in chromosome number groups: 2n = 24 (Aplastodiscus perviridis group), 2n = 22 (Aplastodiscus albofrenatus group), 2n = 20, and 2n = 18 (both within Aplastodiscus albosignatus group). This study aims to report karyotypic information on 4 species from the last 2 groups using classical and molecular cytogenetic techniques and hypothesize chromosomal evolutionary trends within the species groups. Aplastodiscus weygoldti showed 2n = 22; Ag-NOR and FISH 18S rDNA signals were located in the interstitial region of the short arms of chromosome pair 6. Aplastodiscus cavicola, Aplastodiscus sp. 4, and Aplastodiscus sp. 6 showed 2n = 18; Ag-NOR and FISH 18S rDNA bands were located in the terminal region of the long arm of chromosome pair 9. Our results support multiple and independent chromosome fusion events within Aplastodiscus, including a new chromosome fission event. Ag-NOR and FISH 18S rDNA patterns were restricted to the small chromosome pairs, similar to the other species within this genus, and confirm overall chromosome morphology conservation among the genera of Cophomantinae.

A new species of Pimelodella Eigenmann amp; Eigenmann, 1888 (Siluriformes: Heptapteridae) from the Sierra Nevada de Santa Marta, Colombia.

A new species of Pimelodella is described from northern coastal drainages of the Sierra Nevada de Santa Marta (SNSM), Colombia. The new species is diagnosed from all trans-Andean congeners by the following unique combination of characters: head length 22.2-23.7% SL, bony interorbital width 17.6-21.6% HL, maxillary barbels length 53.0-68.3% SL, inner mental-barbels length 14.4-20.0% SL, body width 17.6-21.1% SL, dorsal-fin spine length 8.3-13.7% SL, dorsal-fin base 12.3-14.9% SL, pectoral-fin spine length 12.9-14.5% SL, pectoral-fin spine length without dentations on the distal posterior margin 21.6-29.8% in pectoral-fin spine length, maximum depth of dentations in posterior margin of pectoral-fin spine 1.40-1.68 times in the width of the spine at its base, adipose-fin base length 22.8-26.4% SL, caudal-peduncle depth 9.2-10.5% SL, 40 vertebrae, having a conspicuous paired dark brown stripe on the dorsal surface of body, extending from posterior margin of head to caudal-fin insertion and a wide dark brown midlateral stripe present. The isolated occurrence of this new taxon living in allopatry in coastal drainages of the SNSM could have interesting biogeographic implications for dispersal and vicariance processes of the ichthyofauna from northern South America.

Molecular identification of three species of Oncideres (Coleoptera: Cerambycidae) using RAPD markers.

Twig girdlers have a short adult life and a long larval stage (up to one year in some species). This fact, together with lack of morphological traits to identify the species from the larval stage, poses obstacles for the taxonomic identification and characterization of the many twig girdlers found in the wild. To solve this matter, Random Amplified Polymorphic DNA (RAPD) molecular markers have been applied to identify some insect species and to determine species boundaries. The aim of this study was to identify three species of Oncideres using the RAPD technique. Adults and larvae of O. saga, O. ocularis and O. ulcerosa were collected in Minas Gerais, Brazil. Two RAPD primers were used (OPA-05 and OPB-13), which produced different electrophoretic profiles that were used to construct a UPGMA phenogram. Three groups identified in the analyses accurately separated the three species, based on individuals in two different stages of development (adults and larvae): I: O. saga, II: O. ulcerosa, and III: O. ocularis. Molecular markers, such as RAPD, are valuable tools that help taxonomists in species identification.

Cytogenetic Analysis and Chromosomal Mapping of Repetitive DNA in Melipona Species (Hymenoptera, Meliponini).

Stingless bees of the genus Melipona are subdivided into 4 subgenera called Eomelipona, Melikerria, Melipona sensu stricto, and Michmelia according to species morphology. Cytogenetically, the species of the genus Melipona show variation in the amount and distribution of heterochromatin along their chromosomes and can be separated into 2 groups: the first with low content of heterochromatin and the second with high content of heterochromatin. These heterochromatin patterns and the number of chromosomes are characteristics exclusive to Melipona karyotypes that distinguish them from the other genera of the Meliponini. To better understand the karyotype organization in Melipona and the relationship among the subgenera, we mapped repetitive sequences and analyzed previously reported cytogenetic data with the aim to identify cytogenetic markers to be used for investigating the phylogenetic relationships and chromosome evolution in the genus. In general, Melipona species have 2n = 18 chromosomes, and the species of each subgenus share the same characteristics in relation to heterochromatin regions, DAPI/CMA3 fluorophores, and the number and distribution of 18S rDNA sites. Microsatellites were observed only in euchromatin regions, whereas the (TTAGG)6 repeats were found at telomeric sites in both groups. Our data indicate that in addition to the chromosome number, the karyotypes in Melipona could be separated into 2 groups that are characterized by conserved cytogenetic features and patterns that generally are shared by species within each subgenus, which may reflect evolutionary constraints. Our results agree with the morphological separation of the Melipona into 4 subgenera, suggesting that they must be independent evolutionary lineages.

Phylogeography and Historical Biogeography of the Astyanax bimaculatus Species Complex (Teleostei: Characidae) in Coastal Southeastern South America.

Astyanax bimaculatus, a ubiquitous species in many Neotropical basins, is characterized by a complex taxonomy and are currently considered a species complex. The goal of this study was to analyze 31 populations (N = 136) of this species from southeastern Brazil using cytogenetic techniques: conventional staining, nucleolar organizer region (Ag-NOR), C-banding, and 18S and 5S recombinant DNA fluorescent in situ hybridization (FISH) probes; and molecular techniques: S72, RAG2, and COI. All populations were 2n = 50 (6m + 20sm +18st +6a); Ag-NORs were predominantly simple, C-banding revealed high variation levels within and among basins, and the FISH probes 18S and 5S were restricted to chromosome pairs 14 and 7, respectively. The S72 was uninformative for phylogenetic analyses, and RAG2 showed no variation among populations. The COI gene revealed three haplogroups. The most basal was composed of Pandeiros population (São Francisco Basin) that diverged in the Middle Miocene. The second was composed of A. altiparanae from the Upper Paraná Basin and Espírito Santo Stream (Paraíba do Sul Basin), whereas the third was composed of Astyanax lacustris from São Francisco and coastal basins. The second and third haplogroups diverged in the Pleistocene, indicating that diversification of the bimaculatus complex was driven by tectonic activity and sea-level fluctuations.

Trends in Karyotype Evolution in Astyanax (Teleostei, Characiformes, Characidae): Insights From Molecular Data.

The study of patterns and evolutionary processes in neotropical fish is not always an easy task due the wide distribution of major fish groups in large and extensive river basins. Thus, it is not always possible to detect or correlate possible effects of chromosome rearrangements in the evolution of biodiversity. In the Astyanax genus, chromosome data obtained since the 1970s have shown evidence of cryptic species, karyotypic plasticity, supernumerary chromosomes, triploidies, and minor chromosomal rearrangements. In the present work, we map and discuss the main chromosomal events compatible with the molecular evolution of the genus Astyanax (Characiformes, Characidae) using mitochondrial DNA sequence data, in the search for major chromosome evolutionary trends within this taxon.

The Nothoaspis amazoniensis Complete Mitogenome: A Comparative and Phylogenetic Analysis.

The molecular biology era, together with morphology, molecular phylogenetics, bioinformatics, and high-throughput sequencing technologies, improved the taxonomic identification of Argasidae family members, especially when considering specimens at different development stages, which remains a great difficulty for acarologists. These tools could provide important data and insights on the history and evolutionary relationships of argasids. To better understand these relationships, we sequenced and assembled the first complete mitochondrial genome of Nothoaspis amazoniensis. We used phylogenomics to identify the evolutionary history of this species of tick, comparing the data obtained with 26 complete mitochondrial sequences available in biological databases. The results demonstrated the absence of genetic rearrangements, high similarity and identity, and a close organizational link between the mitogenomes of N. amazoniensis and other argasids analyzed. In addition, the mitogenome had a monophyletic cladistic taxonomic arrangement, encompassed by representatives of the Afrotropical and Neotropical regions, with specific parasitism in bats, which may be indicative of an evolutionary process of cospeciation between vectors and the host.

Ecohydrological modeling and environmental flow regime in the Formoso River, Minas Gerais State, Brazil.

This paper aimed at determining the environmental flow regime in a 1 km stretch of the Formoso River, MG, using River2D model. To carry out the ecohydrological modeling, the following information was used: bathymetry, physical and hydraulic features, and the Habitat Suitability Index for species of the Hypostomus auroguttatus. In the River2D, the Weighted Usable Areas were determined from the average long-term streamflows with percentage from 10% to 100%. Those streamflows were simulated for the later construction of optimization matrices that maximize the habitat area throughout the year. For H. auroguttatus Juvenile, higher values of Weighted Usable Area were associated with the percentage of 60% and 70% of the average long-term streamflows in October and September, respectively. For H. auroguttatus Adult, the highest value of Weighted Usable Area was associated with the percentage of 100% of the average long-term streamflow in September. The environmental flows found for this stretch of the Formoso River varied over the year. The lowest environmental flow was observed in December (2.85 m3 s-1), while the highest was observed in May (4.13 m3 s-1). This paper shows the importance of ecohydrological studies in forming a basis for water resources management actions.

Karyological study of Ololygon tripui (Lourenço, Nascimento and Pires, 2009), (Anura, Hylidae) with comments on chromosomal traits among populations.

To increase the number of cytogenetic characters used in Ololygon tripui systematics, we applied some cytogenetic techniques such as Giemsa, C- and NOR-banding, and fluorescence in situ hybridization (FISH) with 18S rDNA and repetitive microsatellite DNA probes to the study of four populations from Minas Gerais State (southeastern Brazil). All populations showed 2n = 24 and FN = 48, and chromosomal formula 8m + 10sm + 6st. Nucleolar organizing regions (NORs) were located on chromosome pair 6 in all populations, although in the Tripuí locality additional markings were observed on one homologue of chromosome pair 3. These patterns were partially congruent with results obtained using the 18S rDNA FISH probe. The microsatellites repetitive DNA (GA)15 and (CAT)10 probes accumulated predominantly in the terminal region of all chromosomes. Chromosome morphology and Ag-NOR were conserved among populations, a conserved pattern in Ololygon Fitzinger, 1843. Repetitive DNA FISH probes patterns were similar among populations, but they revealed species-specific differences when compared with other species of the genus Ololygon, suggesting that molecular cytogenetics are potentially more informative in karyologically conservative taxa.

The unique karyotype of Henochilus wheatlandii, a critically endangered fish living in a fast-developing region in Minas Gerais State, Brazil.

Henochilus wheatlandii, the only species of this genus, is critically endangered and was considered extinct for over a century. The rediscovery of this fish in 1996 made it possible to study its phylogenetic relationships with other species in the subfamily Bryconinae. The aim of this study was to characterise the karyotype of H. wheatlandii. Standard staining, C-positive heterochromatin and nucleolar organiser region (NOR) banding, chromomycin A(3) staining, and fluorescent in situ hybridisation (FISH) using 5S rDNA and 18S rDNA probes were conducted on nineteen specimens collected in the Santo Antonio River, a sub-basin of the Doce River in Ferros municipality, Minas Gerais State, Brazil. Henochilus wheatlandii shared the same diploid number and chromosome morphology as other species of Bryconinae. However, its heterochromatin distribution patterns, NOR localisation, and FISH patterns revealed a cytogenetic profile unique among Neotropical Bryconinae, emphasizing the evolutionary uniqueness of this threatened species.

Plagioscion squamosissimus (Sciaenidae) and Parachromis managuensis (Cichlidae): a threat to native fishes of the Doce River in Minas Gerais, Brazil.

The middle section of the lake basin of the Doce River in Minas Gerais State, Brazil is plagued by grave environmental problems, including the introduction of non-native fish, which reduces the biodiversity of this region. This study reports the presence of two newly-detected non-native species in the Doce River Basin. Sampling efforts included gill nets with mesh size of 3 to 12 mm (measured diagonally) and trawling nets, both of which were used in independent field campaigns in 2002 and 2011. The two new invasive Perciform fishes, Plagioscion squamosissimus (Heckel 1840) and Parachromis managuensis (Günther 1867) were collected in Caratinga and Rio Doce municipalities. These records and other reports on non-native fishes suggest favorable environmental conditions for the establishment of invasive species in this drainage. These invasive species have behavior and diet observed in other wide distribution exotic fish of Rio Doce Basin representing a threat to the 77 native fishes of this region, 37 of which are endangered.

New Hyphessobrycon from the upper rio Pardo drainage in eastern Brazil (Teleostei: Characiformes: Characidae)

A new species of characid, Hyphessobrycon vinaceus, is described from the upper rio Pardo, Minas Gerais, Brazil. The new species is distinguished from all other Hyphessobrycon species by the combination of the following characters: the body red or reddish pigmented in live specimens, an anterior and conspicuous vertically elongate black humeral spot followed by a second faint humeral spot, 5 teeth in the inner series of premaxilla, maxilla with 2-3 pentacuspidate teeth, 15-17 branched anal-fin rays, 4-5 scale rows above and 3-4 scale rows below lateral line, lateral line with 10-26 perforated scales, 32-34 scales in a longitudinal series, and by a smaller head length in relation to standard length (23.9-26.7 percent).

Uma nova espécie de caracídeo, Hyphessobrycon vinaceus é descrita para o alto rio Pardo, Minas Gerais, Brasil. A nova espécie distingue-se das demais espécies de Hyphessobrycon pela combinação dos seguintes caracteres: coloração avermelhada do corpo em vida, uma mancha umeral preta e conspícua verticalmente alongada seguida de uma segunda mancha umeral fracamente pigmentada, 5 dentes na série interna do pré-maxilar, 2-3 dentes pentacuspidados no maxilar, 15-17 raios ramificados na nadadeira anal, 4-5 escamas acima e 3-4 escamas abaixo da linha lateral, 10-26 escamas perfuradas na linha lateral, 32-34 escamas em uma série longitudinal e menor comprimento da cabeça em relação ao comprimento padrão (23,9-26,7 por cento).

The karyotypes of the thorny catfishes Wertheimeria maculata Steindachner, 1877 and Hassar wilderi Kindle, 1895 (Siluriformes: Doradidae) and their relevance in doradids chromosomal evolution.

We studied the karyotypes of two doradids, the rare and endangered Wertheimeria maculata and a derived Amazonian species, Hassar wilderi. Cytogenetic characterization was assessed using conventional staining (Giemsa), C-banding, and NOR banding. Both species had 2n = 58 chromosomes but differed in their chromosome formulae, 24 m + 14sm + 8st + 12a for W. maculata and 32 m + 16sm + 10st for H. wilderi. In W. maculata heterochromatin was mainly telomeric, and three chromosomes had a fully heterochromatic arm; in H. wilderi heterochromatin was also predominantly telomeric and evident in many more chromosomes. Hassar wilderi also presented one pair of homologues with a fully heterochromatic arm. In both species, nucleolar organizer regions were restricted to one pair of subtelocentric chromosomes. Assuming a basal position for W. maculata, we hypothesized that underlying conserved diploid and NOR-bearing chromosome numbers, chromosomal evolution in doradids has involved pericentric inversions and an increase of heterochromatic blocks.

Phylogeography and RAPD-PCR variation in Hoplias malabaricus (Bloch, 1794) (Pisces, Teleostei) in southeastern Brazil

In the Rio Doce basin of southeastern Brazil, the freshwater fish Hoplias malabaricus (trahira) is a widespread predatory characin and one of the few resilient native fishes in a highly impacted lake system. In order to test for genetic differentiation in populations within this basin and for biogeographic relationships among populations of this species in other basins, a study was conducted using RAPD-PCR analysis of Rio Doce samples (N = 63) and phylogeographic analyses with mitochondrial DNA (mtDNA) haplotypes, including the Rio Grande and Macacu river basins. In the Rio Doce basin, the patterns of genetic similarity of RAPD-PCR markers (individual fingerprinting and NeiÆs genetic distance) suggest the existence of two genetically different groups, one composed of the lacustrine populations Carioca and Dom Helvécio, and the other of riverine and the remaining lacustrine populations. The differences in the RAPD-PCR patterns may be explained by the existence of sub-basins within this lacustrine system. A maximum parsimony tree of cytochrome b fragment (383 base pairs) supports the view that trahiras of the Rio Doce share a complex biogeographic history with those of neighboring basins. The phylogeographic patterns may be explained by a common history of the watersheds of the Rio Doce, Paraíba do Sul, and Rio Grande basins, corroborating the hypothesis of a Plio-Pleistocene separation of these drainage systems, forming the Mantiqueira ôdivortium aquariumö