Research Projects
2020 - 2021
The use of molecular cytogenetics for evolutioonary and taxonomic studies in Cervidae.
Abstract: The family Cervidae groups together species with a growing economic potential and endangered species. Karyotypes and phylogenetic position of many deer species are uncertain due to the lack of detailed scientific data. The extensive variation in chromosome numbers (2n=6-70) and structure makes this family suited for evolutionary studies based on interspecies molecular cytogenetic comparisons. In this project, we will perform a comparative analysis in a wide range of deer species, focusing on species lacking comprehensive data, including the genus Mazama and its cytotypes. We will determine karyotype differences and their impact on production of aneuploid spermatozoa and reproductive isolation, identify intrachromosomal rearrangements of the X chromosomes, and compare sequence similarities and chromosomal positions of satellite DNAs in various deer species. For this purpose, whole chromosome, BAC and satellite DNA probes for FISH will be constructed in VRI laboratory. Obtained data will be used to improve taxonomic classification, and review phylogenetic relationships within Cervidae.
Situation: Ongoing project; Nature: Research.
Student's Participation: Master's: (1) / PhD: (1).
Members: Jose Maurício Barbanti Duarte (Coordinator) / Miluše Vozdová / David Javier Galindo Huamán / Agda Maria Bernegossi.
Funder: São Paulo Research Foundation (FAPESP) - Research grant.
2018 - 2023
Abstract: Brocket deer species, Mazama genus, are known as a group of mammals with important taxonomic issues. Mostly, this is due to a high level of homoplasy of morphological characters and high karyotyping diversity. These characteristics suggests that there are still many species to be described, once karyotyping differences among populations generates post-zygotic reproductive barriers. In order to re-describe the species it is necessary to collect specimens at the type localities where each genus name were previously described in a sense of verifying its genetics and morphological constitution and the presence of chromosomal barriers and molecular structure. For these purposes, specimens will be collected at the type localities (new topotypes) of each of the available name, that are considered today a synonymous of known species. Helped by local teams, about 45 specimens will be collected at 16 countries following a chronological sequences of the publications in order to verify its validity. Skin and testis fragments, frozen in liquid nitrogen, will be collected. Also, samples of liver, kidneys, spleen and muscles will be collected and preserved in absolute ethanol. All the bones will be cleaned and conserved as dried specimens. The skin will be treated with a tannery solution for deposition in scientific collection. The specimens will be evaluated cytogenetically (chromosomal biometry, G band, C band, NOR band, telomeric FISH and chromosomal paint), molecularly (Cytochrome B, D-Loop, ND5, COI, α-LAlb, IL16, MGF) and morphologically (biometry, skull, post-skull, skin and photographs) and compared with patterns previously obtained for the older names. Sperm FISH experiments of gametic segregation will be carried out to evaluate the thresholds of the chromosomal differences that became efficient reproductive barriers. From this point, that threshold will be considered to describe distinct species. Moreover, methodological studies will be developed to evaluate hair samples as a potential non-invasive biological matrix for morphology, cytogenetics and molecular genetics. Also, the species group speciation process will have an ecological approach to understand the micro and macro environmental niche dimensions overlap among well-established species. With the scientific advance intended with this project it is expected to give sustainability to the IUCN threatened categorizations process contributing to unknown or neglected species conservation. Also, it is expected to strengthen the cervidology of Latin America with technology exchange among the established research groups and creation of new groups in countries where this research field is incipient.
Situation: Ongoing project; Nature: Research.
Student's Participation: Undergraduate: (3) / Master's: (3) / PhD: (3).
Members: Jose Maurício Barbanti Duarte (Coordinator) / Alexandre Vogliotti / Renato Caparroz / Susana Gonzalez / Elias Alberto Gutierrez Canelossi / Márcio Leite de Oliveira / Kathia Rivero Guzmán / Rogério Vieira Rossi / Fernando Camargo Passos / Rafael Reyna-Hurtado / Sonia Gallina / Damián Rumiz / José Luis Cartes / Benoit de Thoisy / Marina Fortes / Galo Zapata-Rios.
Funder: São Paulo Research Foundation (FAPESP) - Research grant.
2013 - 2017
The future of conservation of small threatened populations: genomic bank and reintroduction of marsh deer embryos.
Abstract: The marsh deer has been suffering a serious reduction in its habitat, especially related to the construction of hydroelectric power plants that affect the floodplains of large rivers, an environment occupied by the species. The Brazilian electricity matrix, supplied almost exclusively by hydroelectric power, is booming, which will significantly affect the remaining populations of marsh deer in the Pantanal, and should change its IUCN Red List classification from vulnerable to threatened if this scenario is confirmed. Within this context, the safeguarding of the species' genetic material becomes a fundamental point in its conservation process. With the loss of large areas of floodplains, there will be numerous small areas left, which may be occupied by a small number of individuals. Thus, since small populations are seriously affected by drift and inbreeding, this scenario will require population management measures to be taken to maintain the genetic diversity of these small populations. In this context, the Deer Research and Conservation Center (NUPECCE) has, since 1998, been carrying out experiments to monitor and maintain a small reintroduced population of marsh deer in the floodplains adjacent to the Ecological Station of Jatai (EEJ), in São Paulo state, on the banks of the Mogi-Guaçu River, where the species was extinct in the middle of the last century. The population grew from eight reintroduced animals (three males and five females) and in 2007, when it was last monitored, it had around 20 animals. However, studies carried out by NUPECCE, from a genetic point of view, have defined the need for the reintroduction of an animal every four years so that genetic diversity is maintained over generations. It is known that the translocation of animals to a population is a delicate and risky process due to the risk of disease transmission to the recipient population, in addition to the low viability in nature of translocated animals from captive populations. This can lead to failure of the translocation, initially by death of the translocated animal or even due to the inability of this individual to dispute mating. Within this context, the aim of this project is to carry out a marsh deer population estimate in EEJ, as well as to introduce new genetic material into the population through the transfer of embryos, in addition to collecting material from the captive population to set up a cryogenic bank of the species.
Situation: Concluded; Nature: Research.
Student's Participation: Undergraduate: (2) / Master's: (3) / PhD: (4) .
Members: Jose Maurício Barbanti Duarte - Coordinator / Eveline dos Santos Zaneti / Marina Suzuki Cursino / Aline Meira Bonfim Mantellatto / Márcio Leite de Oliveira / Luciana Diniz Rola / Pedro Henrique Faria Peres / Ellen F. C. Peroni / Francisco Grotta Neto / David Javier Galindo Huamán / Gabriella Ferreira.
Funder: Brazilian Biodiversity Fund (FUNBIO) - Research grant.
2013 - 2016
A busca dos paratopótipos e produção de citótipos e genótipos como a melhor forma para resolução da complexa taxonomia do gênero Mazama (Mammalia; Cervidae)
Abstract: Numerous taxonomic problems have been found in the species of the genus Mazama. Recent works have proposed that the gray Mazama do not belong to the genus Mazama and that M. nemorivaga does not share the same genus with M. gouazoubira. On the other hand, M. nemorivaga itself has its validity as a species corroborated by some authors. Still, the existence of innumerable new species within what is now recognized as M. americana is suggested. These advances have been generated by the advent of cytogenetic and molecular techniques, which bring new forms of analysis to the old morphological evaluations of the skulls and skins, which were biased by the great evolutionary convergence of the morphological patterns of these species. Cytogenetics, in particular, has shown that Mazama have speciation centered on deep chromosomal rearrangements and that chromosomal variants should be considered as distinct species. But for new species to be described, there is a need to analyze these characteristics of the holotypes of the species already described, and those that are different from them would be the new species. However, the cytogenetic analysis of the holotypes is impossible, since it needs living cells, while the molecular analysis has great problems because it is old and highly degraded DNA. Thus, the generation of new "holotypes" from paratopotypes (animals collected in the same location as the holotypes) would be a possibility to generate cytotypes (chromosomal characterization of types) and genotypes (molecular characterization of types) of the species already described and their synonyms, because these synonyms could be valid species. Within this context, the present project aims to contribute to this discussion, reviewing the taxonomy of these species, using traditional methods such as morphology, but also more advanced methods such as cytogenetics and molecular genetics, in an attempt to elucidate the evolutionary processes that involved this group.
Situation: Concluded; Nature: Research.
Student's Participation: Undergraduate: (2) / Specialization: (2) / Master's: (1) / PhD: (1) .
Members: Jose Maurício Barbanti Duarte (Coordinator) / Susana Gonzalez / José Eduardo Garcia / Vanessa Veltrini Abril / Aline Meira Bonfim Mantellatto / Iara Maluf Tomazella.
Funder: National Council for Scientific and Technological Development (CNPq) - Research grant.
2013 - 2016
Behavioral interactions and pre-zygotic isolation between populations of the genus Mazama (Mammalia Cervidae): implications for systematics and conservation.
Abstract: The phylogeny of the deer currently positioned in the genus Mazama is under discussion. Recent work indicates that it is a monophyletic group that includes a set of cryptic species, the result of processes of evolutionary convergence that occurred in different strains in their adaptation to the ecological conditions of tropical forests. This project aims to prove experimentally the existence of pre-zygotic reproductive barriers based on sexual behavior and pheromones. For this, sexual recognition experiments will be carried out, as well as chemical analyzes of the compounds present in the urine and glandular secretionsof the animals. The results of the project and the information available on the topic will be used as the basis for a television program that will bring the latest discoveries about the evolution of the genus Mazama and other South American Cervids to the general public, in order to raise awareness of the need to conserve natural areas and threatened populations.
Situation: Concluded; Nature: Research.
Student's Participation: Specialization: (1) / PhD: (1) .
Members: Mar Roldán Romero (postdoc) / Jose Maurício Barbanti Duarte (Coordinator) / Mateus José Rodrigues da Costa / Juán Carranza Almansa.
Funder: Coordination for the Improvement of Higher Education Personnel (CAPES)- Research grant.
2013 - 2015
Abstract: The Brazilian dwarf brocket deer (Mazaman nana) is a cervid species that occupies the forests of Southern Brazil and has been greatly affected by the drastic reduction of forested areas. It is also the neotropical deer less studied by Science. Aware about this situation, this project aims to determine the species basic ecological parameters such as distribution, abundance and sex ratio. Given the rarity and high elusiveness of the species, it is proposed the use of indirect methods to achieve this objective. Thus, will be used methodologies based on the collect of faecal samples, DNA extraction and subsequent molecular and genetic analysis. Faecal samples will be collected with the help from a scat detection dog, in protected areas spread over the south of Brazil. After species identification by PCR/RFLP and specialization of the samples, will be made the species distribution modeling at the software Maxent using the environmental variables from Ambdata database of INPE. A protected area will be choose for a faecal sampling based on transects, every month for three months, in order to estimate the population through models of capture-mark-recapture. With this data will be calculated the density of the species in the area sampled. For this, faeces samples will be individualized with microsatellite markers. The sex ratio will be calculated with the abundance data and the sexing of the samples by the amelogenin locus amplification. This information will contribute to the beginning of an adequate species monitoring program and to determine priority areas for the dwarf brocket deer conservation.
Situation: Concluded; Nature: Research.
Student's Participation: Master's: (1) / PhD: (1).
Members: Jose Maurício Barbanti Duarte (Coordinator) / Aline Meira Bonfim Mantellatto / Márcio Leite de Oliveira / Pedro Henrique Faria Peres.
Funder: São Paulo Research Foundation (FAPESP) - Research grant.
2011 - 2015
Abstract: Many taxonomic problems have been found in the brocket deer (Mazama Genus). Recent work have suggested that gray brockets do not belong to the Mazama genus, and M. nemorivaga (Amazonian gray brocket deer) and M. gouazoubira (Brown brocket deer) also integrate different genera. Meanwhile, M. nemorivaga is not accepted by some researchers as valid species. The present project aims to contribute with this discussion, reviewing the taxonomy of these species using traditional methods, as morphology, and to elucidate the evolution processes of this group through modern methods as cytogenetics and molecular genetics. The project also plans to evaluate the ecological differences between these species. To accomplish these objectives, it will be necessary to develop methodologies for ecological studies using fecal DNA, radiotelemetry and camera traps for these animals. We are planning to obtain a broad sample in Brazilian gray brockets (M. gouazoubira and M. nemorivaga), and some of them will be conducted to the Deer Research and Conservation Center (NUPECCE) captive facilities for reproductive studies. A taxonomic review will be conducted in South American museums analyzing skulls and skins. It will be collected material, in live animals, for cytogenetic and molecular analysis. Cytogenetically, we will compare the banding patterns, C, G, NOR, telomeric FISH and chromosome painting. It will be evaluated two mitochondrial markers (Citochrome b, D-loop or Citochrome Oxidase) and two nuclear markers (to be defined in the future) to show the molecular differences between individuals that will be compared with the Neotropical deer sequences placed in the Genbank. The captive animals will be crossed to produce pure (M. gouazoubira x M. gouazoubira and M. nemorivaga x M. nemorivaga) and hybrid (M. gouazoubira x M. nemorivaga) lineages. The animals produced will be evaluated to access their fertility and define the existence of reproductive isolation factor between these species, and to confirm definitively the validity of the species. Five M. gouazoubira will be captured in the Nhecolandia's Pantanal. These animals will receive GPS tracking collars to establish home range and habitat use pattern. In the region of the captured animals, itwill be used camera traps and fecal DNA to test these methods as valid tools to estimate animal home range. The animals with GPS tracking collar will be used as control. Since the capture of M. nemorivaga is impossible, we will use the fecal DNA and camera traps to evaluate the home range and activity pattern of this species in the Amazonia (Jamari National Forest. We hope to contribute to solve taxonomic problems and consolidate the biologic knowledge of the gray brockets, generating technology for future work with these and similar species.
Situation: Concluded; Nature: Research.
Student's Participation: Master's: (3) / PhD: (3).
Members: Jose Maurício Barbanti Duarte - Coordinator / Susana Gonzalez / José Eduardo Garcia / Artur Andriolo / Ubiratan Piovezan / Mariano Lisandro Merino / Vanessa Veltrini Abril / Eveline dos Santos Zanetti / Allyson Diaz Koester / Aline Meira Bonfim Mantellatto / Jesus Eduardo Maldonado / Vinícius Soares Antunes / Javier Adolfo Sarria-Perea / Christofoletti, Maurício Durante / Marina Gomes de Figueiredo / Josi Cerveira / Rosana Nogueira de Morais.
Funder: São Paulo Research Foundation (FAPESP) - Research grant.
