Search

B chromosomes occur in different species of the small characid fishes of the genus Moenkhausia. These supernumerary elements, that do not recombine with chromosomes of the standard A complement and follow their own evolutionary mechanism vary in number, morphology, and distribution. Here, we show karyotypic data of individuals of 2 populations of Moenkhausia oligolepis of the Brazilian Amazon (Pedro Correia and Taboquinha streams, Tocantins river basin), both with a diploid number of 50 chromosomes and karyotypic formula of 10m + 32sm + 8a. In addition to the normal complement, we also observed the occurrence of B chromosomes in the 2 populations with intra- and interindividual variation ranging from 0 to 10 Bs, independent of sex. The C-banding pattern evidenced heterochromatic blocks located mainly in the pericentromeric region of the chromosomes, while the B chromosomes appeared euchromatic. Silver-stained nucleolus organizer regions were identified in multiples sites, and some of these blocks were positive when stained with chromomycin A3. The karyotype analysis and the application of whole-chromosome painting in populations of M. oligolepis reinforce the conservation of the basal diploid number for the genus, as well as the evolutionary tendency in these fishes to carry B chromosomes. Both populations turned out to be in different stages of stability and expansion of their B chromosomes. We further suggest that the origin of these chromosomes is due to the formation of isochromosomes. Here, we identified a pair of complement A chromosomes involved in this process.

Made available in DSpace on 2020-12-12T01:59:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-06-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The streaked prochilod Prochilodus lineatus is an important fish species from the Neotropical region. In this study, a protocol of triploidization was established by temperature shock. Fertilized eggs were shocked at 2 min post-fertilization at 0, 38, 40 and 42°C. At 0°C, the embryos were maintained for 30 min, while the rest for 2 min. Heat shocked embryos and control (during all experiment) were incubated at 27°C. The ploidy status was confirmed by flow cytometry, erythrocyte diameter, conventional cytogenetics (Giemsa staining), chromosome banding and fluorescence in situ hybridization using 5S and 18S rDNA probes. Heat-shock at 40°C produced 96.7% of triploids and such a procedure did not reduce the fertilization, hatching rate and the percentage of normal embryos. The use of chromosome banding and FISH gave rise to effective procedure to identify triploids. The data obtained here is innovative for the species and bring new information for basic and applied studies. Laboratory of Fish Biotechnology National Center for Research and Conservation of Continental Fish Chico Mendes Institute of Biodiversity Conservation Department of Veterinary Medicine – FZEA University of São Paulo Aquaculture Center São Paulo State University Federal Rural University of Rio de Janeiro Faculdade de Ciências Bauru UNESP – Univ. Estadual Paulista Aquaculture Center São Paulo State University Faculdade de Ciências Bauru UNESP – Univ. Estadual Paulista FAPESP: 2010/17429-1 FAPESP: 2011/11664-1

Made available in DSpace on 2020-12-12T01:56:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-06-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis Billfish are oceanic pelagic species that are often caught by tuna fleets and are of great interest for sport fishing. Two species of billfish have specific legislation prohibiting their marketing and export in Brazil. DNA barcoding is a universal system of molecular identification based on a sequence of mitochondrial DNA cytochrome oxidase subunit I (COI), which serves as a diagnostic genomic marker in each species. The barcode DNA technique was used to identify billfish marketed in the second largest fishing warehouse in Latin America, the CEAGESP (Companhia de Entrepostos e Armazéns Gerais de São Paulo), located in São Paulo, Brazil. Seventy-nine samples of billfish were collected during three inspection visits carried out by Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis. After DNA sequencing, 70 samples (88.60%) were identified to the species level; 21 (30.00%) were identified as Xiphias gladius, 43 (61.42%) as Istiophorus platypterus and six (8.57%) as Kajikia albida. The sale of this latter species is prohibited in Brazil and it is considered Vulnerable on the list of endangered species of the IUCN and in the official list of species of endangered Fauna – Fish and Aquatic Invertebrates. Molecular analyses proved to be very efficient at uncovering irregularities in the identification of the white marlin (K. albida), which was traded illegally in CEAGESP, demonstrating the ineffectiveness of the current monitoring techniques used and emphasizing the need for the adoption of better public policies for the conservation of the species. Departamento de Ciências Biológicas Universidade Estadual Paulista Centro de Aquicultura da Unesp Universidade Estadual Paulista Departamento de Morfologia Universidade Estadual Paulista Departamento de Ciências Biológicas Universidade Estadual Paulista Centro de Aquicultura da Unesp Universidade Estadual Paulista Departamento de Morfologia Universidade Estadual Paulista

DNA Barcode were used to identify species Billfishes and swordfish Xiphias gladius appeared as the only billfish species exported in Brazil. This system has been used successfully in the identification of species and samples not characterized, or illegally traded, as well as products from threatened species. The DNA barcode technique was used in the identification of billfish products exported through the terminal of the International Airport of Sao Paulo. Samples of billfish tissues were collected during the procedures to export the products to the United States and England. DNA sequencing permitted to certify that 100 % of the sampled products inspected were correctly identified as Xiphias gladius, thus confirming the NCM information about the species declared by the exporter. The high index of similarity of the sequences obtained in the samples with the sequences deposited in the Bold Systems and GenBank which is an NCBI platform (National Center for Biotechnology Information) platform for this species allow the validation and adequacy of the technique used, with direct application mainly in the process of inspection of the commercialization and exportation of fishery products.

Made available in DSpace on 2022-04-29T08:36:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) B chromosomes are non-essential additional genomic elements present in several animal and plant species. In fishes, species of the genus Psalidodon (Characiformes, Characidae) harbor great karyotype diversity, and multiple populations carry different types of non-essential B chromosomes. This study analyzed how the dispensable supernumerary B chromosome of Psalidodon paranae behaves during meiosis to overcome checkpoints and express its own meiosis-specific genes. We visualized the synaptonemal complexes of P. paranae individuals with zero, one, or two B chromosomes using immunodetection with anti-medaka SYCP3 antibody and fluorescence in situ hybridization with a (CA)15 microsatellite probe. Our results showed that B chromosomes self-pair in cells containing only one B chromosome. In cells with two identical B chromosomes, these elements remain as separate synaptonemal complexes or close self-paired elements in the nucleus territory. Overall, we reveal that B chromosomes can escape meiotic silencing of unsynapsed chromatin through a self-pairing process, allowing expression of their own genes to facilitate regular meiosis resulting in fertile individuals. This behavior, also seen in other congeneric species, might be related to their maintenance throughout the evolutionary history of Psalidodon. Universidade Estadual Paulista (UNESP) Instituto de Biociências de Botucatu Departamento de Biologia Estrutural e Funcional Universidad de La Serena Instituto de Investigación Multidisciplinar en Ciencia y Tecnología Universidad de La Serena Departamento de Biología Laboratorio de Genética y Citogenética Vegetal Hokkaido University Faculty of Science Department of Biological Sciences Laboratory of Reproductive & Developmental Biology Universidade Estadual Paulista (UNESP) Faculdade de Ciências Departamento de Ciências Biológicas Universidade Federal de São Carlos (UFSCAR) Departamento de Genética e Evolução Universidade Estadual de Ponta Grossa (UEPG) Departamento de Biologia Estrutural Molecular e Genética Universidade Estadual Paulista (UNESP) Instituto de Biociências de Botucatu Departamento de Biologia Estrutural e Funcional Universidade Estadual Paulista (UNESP) Faculdade de Ciências Departamento de Ciências Biológicas FAPESP: 2013/24143-5 FAPESP: 2014/09634-5 FAPESP: 2014/26508-3 FAPESP: 2016/09204-6 FAPESP: 2017/15608-5 FAPESP: 2017/22447-8 FAPESP: 2018/20610-1 CNPq: 306054/2006-0

Made available in DSpace on 2022-04-29T08:34:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-12-01 Comisión Nacional de Investigación Científica y Tecnológica Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Scarce genomic resources have limited the development of breeding programs for serrasalmid fish Colossoma macropomum (tambaqui) and Piaractus mesopotamicus (pacu), the key native freshwater fish species produced in South America. The main objectives of this study were to design a dense SNP array for this fish group and to validate its performance on farmed populations from several locations in South America. Using multiple approaches based on different populations of tambaqui and pacu, a final list of 29,575 and 29,612 putative SNPs was selected, respectively, to print an Axiom AFFYMETRIX (THERMOFISHER) SerraSNP array. After validation, 74.17% (n = 21,963) and 71.25% (n = 21,072) of SNPs were classified as polymorphic variants in pacu and tambaqui, respectively. Most of the SNPs segregated within each population ranging from 14,199 to 19,856 in pacu; and from 15,075 to 20,380 in tambaqui. Our results indicate high levels of genetic diversity and clustered samples according to their hatchery origin. The developed SerraSNP array represents a valuable genomic tool approaching in-depth genetic studies for these species. São Paulo State University (Unesp) Aquaculture Center of Unesp Facultad de Ciencias Veterinarias y Pecuarias Universidad de Chile Facultad de Zootecnia Universidad Nacional Agraria de la Selva Facultad de Ciencias Agrarias Universidad de Antioquia São Paulo State University (Unesp) Aquaculture Center of Unesp CNPq: 140740/2016–3 FAPESP: 2016/18294–9 FAPESP: 2016/21011–9 FAPESP: 2017/19717-3 CNPq: 311559/2018–2

Made available in DSpace on 2021-06-25T10:59:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Since the Miocene profound climatic changes have influenced the biology and ecology of species worldwide, such as their connectivity, genetic population structure, and biogeography. The goal herein is to evaluate the phylogeography of sailfish Istiophorus platypterus between the Atlantic, Indian, and Pacific oceans. Our results evidenced a high genetic diversity and three distinct populations among the ocean basins with limited gene flow among them. In addition, the species is characterized by two deep evolutionary lineages that diverged during the Miocene/Pliocene transition, one of them is circumtropical while the other is restricted to the Atlantic Ocean. These lineages evolved along the successive glacial-interglacial cycles from the Pleistocene and remained isolated from each other in glacial refugium until deglaciation. Assessments of sailfish suggest it may be subject to overfishing and the results herein imply the need to re-evaluate the current stock delimitations and management measures adopted by the Regional Fisheries Management Organizations, especially in the Atlantic and the Indo-Western Pacific oceans to effectively manage the species. In addition, this work highlights that both lineages should at least be treated as two distinct management units in the Atlantic Ocean until their taxonomic status is fully resolved, given their high genetic divergence. Conservation Genetics Laboratory Universidade Santa Cecília (UNISANTA), 8, Cesário Mota St. Laboratório de Genética Pesqueira e Conservação (GenPesC) Instituto do Mar (IMar) Universidade Federal de São Paulo (UNIFESP), Campus Baixada Santista, 144, Dr. Carvalho de Mendonça St. Laboratório de Ciências da Pesca (LABPESCA) Instituto do Mar (IMar) Universidade Federal de São Paulo (UNIFESP), Campus Baixada Santista, 144, Dr. Carvalho de Mendonça St. Laboratório de Oceanografia Pesqueira Universidade Federal Rural de Pernambuco (UFRPE) Instituto Oceanográfico de Venezuela Universidad de Oriente (UDO) School of Biological Sciences The University of Queensland Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis (IBAMA) Laboratório de Genética de Peixes (LAGENPE) Faculdade de Ciências Universidade Estadual Paulista “Júlio de Mesquita Filho” (UNESP) Centro de Pesquisa do Pescado Marinho Instituto de Pesca/APTA/SAA/SP Acervo Zoológico Universidade Santa Cecília (UNISANTA), Oswaldo Cruz St. 266 Instituto Português do Mar e da Atmosfera (IPMA), Av. 5 de Outubro s/n° Centro de Ciências do Mar (CCMAR) Universidade do Algarve, Campus de Gambelas Cooperative Institute for Marine and Atmospheric Studies Rosenstiel School for Marine and Atmospheric Science University of Miami Centre de Recherches Océanographiques de Dakar‐Thiaroye (CRODT) Institut Sénégalais de Recherches agricoles (ISRA) Centre de Recherches Océanologiques (CRO) Centre Technique de Recherche et de Valorisation des Milieux Aquatiques (CAP RUN – CITEB), Magasin 10, Port Ouest Laboratório de Biologia e Genética de Peixes (LBGP) Instituto de Biociências de Botucatu (IBB) Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP) Laboratório de Genética de Peixes (LAGENPE) Faculdade de Ciências Universidade Estadual Paulista “Júlio de Mesquita Filho” (UNESP) Laboratório de Biologia e Genética de Peixes (LBGP) Instituto de Biociências de Botucatu (IBB) Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP)

Sequences of the mitochondrial gene COI (DNA Barcode) were used to identify species of rays and skates commercialized in fishmongers in Brazil. The comparisons of the obtained sequences with previously published data available in NCBI and BOLD showed that the fish products corresponded to four species, Hypanus dipterurus, Potamotrygon motoro, Paratrygon ajereba and Gymnura altavela, the last of which is classified as vulnerable according to the IUCN Red List and therefore should not be marketed in accordance with the MMA ordinance 445/2015., Highlights • Genetic markers used in the identification of fishery products. • Methods showed effective to identified fish species. • Marketing of fishery products with changed identity. • The control of commercialized rays can be done by applying the DNA barcode technique. • DNA Barcode was applied in the genetic identification of endangered species.

Made available in DSpace on 2019-10-06T16:42:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-08-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Satellite DNAs (satDNAs) are tandemly repeated DNA sequences with great abundance in eukaryotic genomes. A single species may carry up to hundreds of satDNA families, which is collectively called as satellitome, each showing its own dynamics and evolution rates. In this context, all live species contain a satDNA library that may be partially or totally shared with other related species/populations. In the late few years, next-generation sequencing (NGS) and novel bioinformatic tools facilitated the massive characterization of these sequences at low costs, and consequently, comparing satDNAs between species. In this study, we characterized two novel satDNAs (MsaSat03-80 and MsaSat04-142) in three characid fish (Astyanax paranae and Astyanax fasciatus and two populations of Moenkhausia sanctaefilomenae) and mapped their chromosomal location to unveil the evolutionary dynamics of satDNA repeats in those species. Our results evidenced that MsaSat03 is present in the genomes of all analyzed species, but is clustered only in the chromosomes of M. sanctaefilomenae, exhibiting a conserved number and location of sites. Conversely, MsaSat04 sequences is restricted to M. sanctaefilomenae and shows a differential distribution between the two analyzed populations. Altogether, our analyses point to a complex history of satDNA families in characid fish and the utility of NGS data for comparative satDNA analysis. Departamento de Ciências Biológicas Faculdade de Ciências Universidade Estadual Paulista - UNESP Campus de Bauru Departamento de Morfologia Instituto de Biociências Universidade Estadual Paulista - UNESP Departamento de Ciências Biológicas Faculdade de Ciências Universidade Estadual Paulista - UNESP Campus de Bauru Departamento de Morfologia Instituto de Biociências Universidade Estadual Paulista - UNESP

Made available in DSpace on 2021-06-25T10:56:04Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01. Added 1 bitstream(s) on 2021-07-15T15:05:30Z : No. of bitstreams: 1 S1415-47572021000100202.pdf: 411345 bytes, checksum: 0b1ae738cdd91d607d8f321655db7dcb (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The main objective of this study was to estimate the genetic diversity levels and haplotype traceability in pacu Piaractus mesopotamicus from the breeding program located in Brazil by analyses of the mitochondrial DNA control region (mtDNA). Moreover, broodstocks from eight commercial fish farms were used for comparative evaluation, four from Brazil (Br1-Br4) and four from Argentina (Ar1-Ar4). The descriptive results revealed 47 polymorphic sites and 51 mutations, which evidenced 34 haplotypes. Ten haplotypes were shared among fish farms and 24 were exclusive. The nucleotide diversity (π) ranged from 0.00031 to 0.01462 and haplotype diversity (Hd) from 0.125 to 0.868. The analysis of molecular variance (AMOVA) indicated high structure present in the analyzed stocks (FST = 0.13356 and ФST = 0.52707). The genetic diversity was high in most of the commercial broodstocks, especially those from Brazil. We observed seven haplotypes in the genetic breeding population, of which four were exclusive and three shared among the commercial fish farms. The genetic diversity was moderate (π = 0.00265 and Hd = 0.424) and considered appropriated for this breeding population of pacu. Our results provide support for the genetic diversity maintenance and mtDNA traceability of pacu commercial broodstocks. Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP) Centro de Aquicultura Universidad Nacional de Rosario Facultad de Ciencias Bioquímicas y Farmacéuticas-Ministerio de Ciencia Tecnología e Innovación productiva de Santa Fe Centro Científico y Tecnológico Acuario del Río Paraná Universidade Federal de São Paulo (UNIFESP) Instituto do Mar Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP) Faculdade de Ciências Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP) Centro de Aquicultura Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP) Faculdade de Ciências CAPES: 001 FAPESP: 15/14185-8 FAPESP: 2014/03772-7 CNPq: 311559/2018-2 CNPq: 446779/2014-8

Made available in DSpace on 2021-06-25T11:09:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01. Added 1 bitstream(s) on 2021-07-15T15:05:37Z : No. of bitstreams: 1 S0103-84782021000100403.pdf: 820089 bytes, checksum: f46eadd0b745e0e457b3cab89f2b0204 (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) China Three Gorges Corporation Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Leporinus friderici is a migratory neotropical fish with elevated ecological and economic importance in Brazil. Microsatellite markers are highly important in population genetic studies, management, and conservation programs; however, no markers are available for this species. In this study, seven microsatellite loci, previously developed for Megaleporinus obtusidens, were successfully cross-amplified in L. friderici. Among these loci, five presented moderate to high genetic variability levels, with four to seven alleles per loci and expected heterozygosities varying from ≥ 0.574 to 1.000. These markers represent a valuable tool for the future management and ecological studies involving this species and group of neotropical fishes. Faculdade de Ciências Universidade Estadual Paulista (UNESP) Instituto de Biociências Universidade Estadual Paulista (UNESP) Departamento de Genética Instituto de Ciências Biológicas e Saúde (ICBS) Universidade Federal Rural do Rio de Janeiro (UFRRJ) Faculdade de Ciências Universidade Estadual Paulista (UNESP) Instituto de Biociências Universidade Estadual Paulista (UNESP)

Made available in DSpace on 2021-06-25T10:54:44Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-02-03 Eukaryotic genomes contain large amounts of repetitive DNA sequences, such as tandemly repeated satellite DNAs (satDNAs). These sequences are highly dynamic and tend to be genus- or species-specific due to their particular evolutionary pathways, although there are few unusual cases of conserved satDNAs over long periods of time. Here, we used multiple approaches to reveal that an satDNA named CharSat01-52 originated in the last common ancestor of Characoidei fish, a superfamily within the Characiformes order, ∼140-78 Ma, whereas its nucleotide composition has remained considerably conserved in several taxa. We show that 14 distantly related species within Characoidei share the presence of this satDNA, which is highly amplified and clustered in subtelomeric regions in a single species (Characidium gomesi), while remained organized as small clusters in all the other species. Defying predictions of the molecular drive of satellite evolution, CharSat01-52 shows similar values of intra- and interspecific divergence. Although we did not provide evidence for a specific functional role of CharSat01-52, its transcriptional activity was demonstrated in different species. In addition, we identified short tandem arrays of CharSat01-52 embedded within single-molecule real-time long reads of Astyanax paranae (536 bp-3.1 kb) and A. mexicanus (501 bp-3.9 kb). Such arrays consisted of head-to-tail repeats and could be found interspersed with other sequences, inverted sequences, or neighbored by other satellites. Our results provide a detailed characterization of an old and conserved satDNA, challenging general predictions of satDNA evolution. Departamento de Ciências Biológicas Faculdade de Ciências Universidade Estadual Paulista UNESP Campus de Bauru Departamento de Biologia Estrutural e Funcional Instituto de Biociências de Botucatu Universidade Estadual Paulista UNESP Department of Organismal Biology-Systematic Biology Evolutionary Biology Centre Uppsala University Departamento de Genética Instituto de Ciências Biológicas e da Saúde ICBS Universidade Federal Rural do Rio de Janeiro Rio de Janerio Departamento de Ciências Biológicas Faculdade de Ciências Universidade Estadual Paulista UNESP Campus de Bauru Departamento de Biologia Estrutural e Funcional Instituto de Biociências de Botucatu Universidade Estadual Paulista UNESP

Made available in DSpace on 2020-12-12T02:41:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-06-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Among several marine pelagic species of the Brazilian coast, Scomber colias Gmelin, 1789 (Perciformes: Scombridae) stands out for having great economic importance, since it is widely used as a food resource and presents moderate vulnerability. Twenty specimens of S. colias were purchased from October 2015 to October 2016 from the coast of Santa Catarina, Brazil. In the present study, we recorded Hysterothylacium fabri (Rudolphi, 1819) (Deardorff and Overstreet, Proc Biol Soc Wash 93(4):1035–1079 1980) from the S. colias intestine using an integrative taxonomy approach, where morphological data are used in combination with partial sequences of the ITS gene, to validate the taxonomic status of the species and establish their relationships with other members of the genus. This species is being recorded for the first time in the South Atlantic and S. colias. The specimens of H. fabri collected in this study parasitizing S. colias presented morphology similar to the other specimens already registered parasitizing other hosts. The distance matrix generated showed that the partial sequences obtained in this study were more similar to sequences of Hysterothylacium sp. collected in China. In phylogenetic analysis, the two detected haplotypes of this study were grouped with H. fabri haplotypes deposited in GenBank in a monophyletic subclade. Departament of Parasitology Universidade Estadual Paulista “Júlio de Mesquita Filho” – UNESP Departament of Genetics Faculty of Veterinary Medicine University of Santiago de Compostela – USC Department of Morphology Institute of Biosciences Universidade Estadual Paulista “Júlio de Mesquita Filho” – UNESP Postgraduate Program in Environmental Systems Analysis Centro Universitário Cesmac – CESMAC Departament of Parasitology Universidade Estadual Paulista “Júlio de Mesquita Filho” – UNESP Department of Morphology Institute of Biosciences Universidade Estadual Paulista “Júlio de Mesquita Filho” – UNESP FAPESP: 2015/00207-0

Made available in DSpace on 2020-12-10T20:01:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-05-27 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Astyanax Baird et Girard, 1854, is one of the largest genera in the family Characidac and comprises 177 valid species. This genus has been the focus of cytogenetic studies primarily owing to the presence of B chromosomes and high karyotypic diversity among different populations. The intense genetic variability in Astyanax is one of the factors responsible for the occurrence of species complexes, which are groups (1) with certain difficulties in establishing common genetic pools or (2) belonging to different cryptic species. To evaluate cytogenetic marker inheritance and the possibility of the identification of these hybrids, this study aimed to describe cytogenetic hybrids from three strains of species of the genera Astyanax and Hyphessobrycon Eigenmann, 1908. A. lacustris Lutken, 1875, A. schubarti Britski, 1964, A. fasciatus Cuvier, 1819, and H. anisitsi Eigenmann, 1907 were used to generate three hybrid lineages. The diploid number, heterochromatin sites, and ribosomal genes (18S and 5S rDNA) of the parental strains and the hybrids were analyzed. The results indicated that the three hybrid lineages had cytogenetic markers of both parents, presenting Mendelian inheritance. However, differences in distribution of heterochromatic blocks were observed between the hybrids and the parent strains. Our results allowed the identification of the hybrid strains based on the cytogenetic markers applied, reinforcing the efficiency of cytogenetic markers as tools for identification and indicating that such events may increase the karyotypic diversity in the genera Astyanax and Hyphessobrycon. Univ Estadual Paulista UNESP Julio de Mesquita Fi, Fac Ciencias, Edmundo Carrijo Coube Ave, Bauru, SP, Brazil Ctr Nacl Pesquisa & Conservacao Biota Aquat Conti, Prefeito Uberto Nemesio Pereira Godoi Highways, Pirassununga, SP, Brazil Univ Estadual Ponta Grossa, Setor Ciencias Biol & Saude, Santos Andrade Sq, Ponta Grossa, Parana, Brazil Univ Estadual Paulista UNESP Julio de Mesquita Fi, Ctr Aquicultura UNESP, Prof Paulo Donato Castelane Acess Way, Jaboticabal, SP, Brazil Univ Estadual Paulista UNESP Julio de Mesquita Fi, Inst Biociencias, Prof Montenegro Ave, Botucatu, SP, Brazil Univ Estadual Paulista UNESP Julio de Mesquita Fi, Fac Ciencias, Edmundo Carrijo Coube Ave, Bauru, SP, Brazil Univ Estadual Paulista UNESP Julio de Mesquita Fi, Ctr Aquicultura UNESP, Prof Paulo Donato Castelane Acess Way, Jaboticabal, SP, Brazil Univ Estadual Paulista UNESP Julio de Mesquita Fi, Inst Biociencias, Prof Montenegro Ave, Botucatu, SP, Brazil FAPESP: 2015/12902-4

O objetivo deste estudo foi neutralizar a adesividade de ovos de Pseudoplatystoma fasciatum a partir da utilização de soluções de ureia por 10, 30, 60 ou 90 min (tratamentos T0 a T3) ou em ureia por 10 min e lavados em tanino (T4). No controle, os ovos foram mantidos apenas em água. Após dois experimentos realizados, no grupo controle foram observadas as melhores taxas de viabilidade embrionária, mesmo mantendo a adesividade dos ovos, sendo melhor que os demais tratamentos. No T4 foram verificadas as piores taxas de viabilidade embrionária. Foi observado que os embriões do tratamento T4 apresentaram crescimento reduzido e formaram um grupo separado após análise dos parí­¢metros morfológicos (dií­¢metro do ovo, área total do ovo, área do embrião e área do saco vitelí­­nico) por análise multivariada. Concluiu-se que o grupo controle, livre de produtos quí­­micos, proporcionou os melhores resultados, sendo considerado a melhor alternativa para a conservação imediata e produção aquí­­cola de P. fasciatum.

The aim of this study is to report the presence of a three non-native hybrid long-whiskered catfishes (family Pimelodidae) in the Upper Parana River basin, Brazil. Genetic analyses demonstrated that the three presumptive hybrids were a result of the crossbreeding of Pseudoplatystoma reticulatum (central Amazonas River basin and Lower Parana River) and Leiarius marmoratus (Amazonas, Essequibo and Orinoco rivers), producing a hybrid commonly known in Brazil as cachandia. The potential threat to biodiversity, due to possible genetic contamination, competition and predation of wild stocks, of such artificially produced hybrid fishes is discussed.

Genetic diversity and structure of Pseudoplatystoma corruscans and P. reticulatum, large migratory South America catfishes, where overfishing and the construction of numerous dams in their feeding and reproducing areas are affecting their migratory processes negatively, were studied using microsatellites in samples from Paraguay (that comprises the Pantanal biome), and the upper and lower Parana Basins. Genetic diversity was in accordance to that observed for other large migratory fishes, but the most geographically isolated populations of P. reticulatum and those P. corruscans subject to anthropogenic effects (stocking and dams) showed lower genetic diversity and evidences of bottlenecks compatible with low effective population size. Pseudoplatystoma reticulatum presented subtle genetic differentiation within the Paraguay area, especially between the edges of its distribution. Pseudoplatystoma corruscans, in this same area, presented a quite homogeneous but significant genetic break between the Paraguay and upper Parana populations, apparently resulting from natural and historical isolation between the basins until recently. These data demonstrates that, although these Pseudoplatystoma spp. are abundant in the Pantanal area, anthropogenic events are leading to negative effects on their populations, particularly in the upper Parana Basin. Genetic differentiation observed along each species distribution demands conservation actions to preserve each population's biodiversity. These results represent important genetic information using new microsatellite markers and the first genetic study of P. reticulatum covering this area of its native distribution. Data may also contribute to a better understanding of species migration patterns and to be used as a baseline for proper management.

Made available in DSpace on 2022-05-01T06:02:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-06-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) B or supernumerary chromosomes are dispensable elements that are widely present in numerous eukaryotes. Due to their non-recombining nature, there is an evident tendency for repetitive DNA accumulation in these elements. Thus, satellite DNA plays an important role in the evolution and diversification of B chromosomes and can provide clues regarding their origin. The characiform Prochilodus lineatus was one of the first discovered fish species bearing B chromosomes, with all populations analyzed so far showing one to nine micro-B chromosomes and exhibiting at least three morphological variants (Ba, Bsm, and Bm). To date, a single satellite DNA is known to be located on the B chromosomes of this species, but no information regarding the differentiation of the proposed B-types is available. Here, we characterized the satellitome of P. lineatus and mapped 35 satellite DNAs against the chromosomes of P. lineatus, of which six were equally located on all B-types and this indicates a similar genomic content. In addition, we describe, for the first time, an entire population without B chromosomes. Faculty of Sciences São Paulo State University Department of Structural and Functional Biology Institute of Biosciences São Paulo State University Mato Grosso State University Department of Genetics Institute of Biological Sciences and Health Rural Federal University of Rio de Janeiro Faculty of Sciences São Paulo State University Department of Structural and Functional Biology Institute of Biosciences São Paulo State University FAPESP: 2018/03365-3 FAPERJ: 211.475/2019 CNPq: 405334/2018-4

The North American channel catfish Ictalurus punctatus Rafinesque 1818 is cultivated in the United States, Asia and Brazilian fish farms, and also utilized as a model species in aquaculture and genetic studies. In this work, cytogenetic analysis of . punctatus from Brazilian aquaculture revealed for the first time the presence of extra elements (supernumerary or B chromosomes) in this species. These elements were characterized as dot-like micro B chromosomes and were found in three individuals (varying from 0 to 1) and in one individual with higher incidence per cell (varying from 0 to 5; mean number of Bs per cell = 2.01). More specific cytogenetic techniques in this individual revealed 58 A chromosomes (standard complement) containing heterochromatic bands in the centromeric regions, a single Ag-NOR in a subtelocentric pair (also positive for 18S rDNA using the FISH technique) and multiple 5S rDNA clusters in three different subtelocentric chromosomes. Four B chromosomes were entirely Ag-NOR positive (also fully heterochromatic) and three presented 18S rDNA clusters by FISH. The occurrence of Ag-NOR and 18S ribosomal genes in both A and B chromosome complements may indicate an intraspecific origin for these extra chromosomes. Additionally, the terminal location of 18S ribosomal clusters in the Ag-NOR-bearing chromosomes and the presence of active NOR in the B chromosomes suggested that breakage events may be related to a possible recent origin of these extra elements. We suggest this data may be useful as cytogenetic information for future elucidation of the composition, origin and evolution of extra chromosomes in fishes.

The pacu (Piaractus mesopotamicus) is a Neotropical fish with remarkable productive performance for aquaculture. Knowledge of genetic resources in Neotropical fish is essential for their applications in breeding programs. The aim of this study was to characterize the genetic diversity of seven farmed populations of pacu which will constitute the basis for a broodstock foundation for coming breeding programs in Brazil. Analysis of one wild population (Paran&aacute, River) was used as a reference to compare genetic parameters in the farmed populations. The analyses were performed using 32 single-nucleotide polymorphisms (SNP) and 8 simple sequence repeat (SSR) markers. No significant differences in genetic diversity between populations estimated through the number of alleles and allelic richness, observed heterozygosity, expected heterozygosity, and minimum allele frequency were detected (p &gt, 0.05). Low genetic diversity was observed in all farmed stocks and the wild population. Moreover, we detected low genetic structure when comparing farmed and wild populations for SNPs (FST = 0.07, K = 3) and SSRs (FST = 0.08, K = 2). Analysis of molecular variance (AMOVA) demonstrated that genetic variation was mostly within populations. Kinship analysis showed that most fish farms included related individuals at a proportion of at least 25%. Our results suggest that the basal broodstock for pacu breeding programs should be founded with individuals from different fish farms for higher genetic diversity and to avoid inbreeding risks.

Made available in DSpace on 2020-12-10T19:41:50Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-07-03 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Complete mitochondrial genomes of the characiform fishes Astyanax fasciatus, Astyanax altiparanae, Hoplias malabaricus (Karyomorph A) and the Gymnotiformes species Gymnotus sylvius and Gymnotus cuia were characterized in the present study. The whole mitogenomes varied from 16,400bp (A. fasciatus) to 17,730 bp (A. altiparanae) long and all of them consisted of 13 protein-coding genes, 22 tRNAs, 2 rRNAs genes, a control region, and origin of light-strand replication. The gene order was similar among all the analyzed species. The nucleotide content of all mitogenomes was also similar, with 29.58-30.95% for A, 27.02-28.65% for T, 26.29-29.99% for C, and 14.41-15.67% for G. Univ Estadual Paulista, Fac Ciencias, Dept Ciencias Biol, Bauru, SP, Brazil Univ Estadual Paulista, Inst Biociencias, Dept Morfol, Botucatu, SP, Brazil Univ Fed Rural Rio de Janeiro, Dept Genet, Seropedica, Brazil Univ Estadual Paulista, Fac Ciencias, Dept Ciencias Biol, Bauru, SP, Brazil Univ Estadual Paulista, Inst Biociencias, Dept Morfol, Botucatu, SP, Brazil FAPESP: 2018/03365-3 CNPq: 405334/2018-4

Made available in DSpace on 2019-10-06T16:25:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-12-01 The accumulation of repetitive DNA sequences on the sex-limited W or Y chromosomes is a well-known process that is likely triggered by the suppression of recombination between the sex chromosomes, which leads to major differences in their sizes and genetic content. Here, we report an analysis conducted on the satellitome of Megaleporinus macrocephalus that focuses specifically on the satDNAs that have been shown to have higher abundances in females and are putatively located on the W chromosome in this species. We characterized 164 satellite families in M. macrocephalus, which is, by far, the most satellite-rich species discovered to date. Subsequently, we mapped 30 satellites, 22 of which were located on the W chromosome, and 14 were shown to exist only on the W chromosome. Finally, we report two simple, quick and reliable methods that can be used for sex identification in M. macrocephalus individuals using fin clips or scales, which could be applicable to future studies conducted in the field of aquaculture. Departamento de Morfologia Instituto de Biociências Universidade Estadual Paulista - UNESP, Distrito de Rubião Junior, s/n Departamento de Genética Universidad de Granada Departamento de Ciências Biológicas Faculdade de Ciências Universidade Estadual Paulista - UNESP, Campus de Bauru CAUNESP Universidade Estadual Paulista - UNESP Campus Jaboticabal Departamento de Morfologia Instituto de Biociências Universidade Estadual Paulista - UNESP, Distrito de Rubião Junior, s/n Departamento de Ciências Biológicas Faculdade de Ciências Universidade Estadual Paulista - UNESP, Campus de Bauru CAUNESP Universidade Estadual Paulista - UNESP Campus Jaboticabal

Made available in DSpace on 2019-10-06T16:43:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 Fundação de Amparo à Pesquisa do Estado do Amazonas Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Arapaima, pirarucu or paiche (Arapaima gigas) is one of the largest freshwater fish in the world, and has a long history of commercial exploitation in the Amazon region. To estimate levels of genetic variability and historical and recent connectivity in Arapaima, we examined variation in eleven microsatellite DNA markers in individuals from 22 localities in Brazil, Colombia, and Peru. The results of analysis of molecular variance, Bayesian clustering and discriminant analysis of principal components showed that Arapaima in our samples represents two major populations, one in the Amazonas and one in the Araguaia-Tocantins River basins. The Amazonas population is further structured by isolation-by-distance with the hydrologically largely unconnected Amapá locality representing the Eastern-most extreme of this continuum; gene flow predominates at distances of less than 1500 km with localities separated by over 2000 km dominated by genetic drift and effectively forming different populations. We saw no evidence of multiple species of Arapaima in the Amazonas basin, and analysis of pairwise genetic divergence (FST) with Mantel tests and correlograms indicated that this largest population exhibits a large-scale pattern of isolation-by-distance, with which results from MIGRATE-N agreed. The degree and significance of genetic divergence indicates that most sampled localities represent demographically independent sub-populations, although we did identify several recent migration events between both proximal and more distant localities. The levels of genetic diversity were heterogeneous across sites, including low genetic diversity, effective population sizes, and evidence of genetic bottlenecks in several places. On average the levels of gene diversity and rarefied allelic richness were higher for localities along the Amazonas mainstem than in the tributaries, despite these being the areas of highest fishing pressure, while the lowest values were found in tributary headwaters, where landscape modification is a significant threat. We recommend that managers consider the regional and local threats to these populations and tailor strategies accordingly, strategies which should ensure the ability of young A. gigas to disperse through floodplain corridors to maintain genetic diversity among otherwise sedentary adult subpopulations. Laboratório de Evolução e Genética Animal/LEGAL Universidade Federal do Amazonas (UFAM) Department of Ichthyology California Academy of Sciences Departamento de Ecologia Universidade Federal do Rio Grande do Norte (UFRN) Acqua Consultoria Ambiental Laboratório de Biologia e Genética de Peixes Instituto de Biociências Universidade Estadual Paulista (UNESP) Departamento de Ciências Biológicas Faculdade de Ciências Universidade Estadual Paulista (UNESP) Campus de Bauru Instituto de Estudos Costeiros Universidade Federal do Pará Campus Universitário de Bragança Laboratório de Biologia e Genética de Peixes Instituto de Biociências Universidade Estadual Paulista (UNESP) Departamento de Ciências Biológicas Faculdade de Ciências Universidade Estadual Paulista (UNESP) Campus de Bauru Fundação de Amparo à Pesquisa do Estado do Amazonas: 062.01341/2018 Fundação de Amparo à Pesquisa do Estado do Amazonas: 899/03 CNPq: CNPq 482662/2013-1 CNPq: CNPq/CT-Amazonia 554057/2006-9 CNPq: CNPq/CT-Amazonia 575603/2008 CNPq: CNPq/MPA 42/2012 CNPq: MCT/CNPq/PPG7 557090/2005-9

This work was carried out to predict the combining abilities, both general and specific, for performance traits and bodily yields of Serrasalmidae. Ninety‐six 30‐day‐old juveniles were purchased from two commercial fish farms, 12 of each of the following eight genetic groups: pacu, pirapitinga, tambaqui, tambacu, tambatinga, patinga, paqui and piraqui. Six fish from each genetic group were grown in 500‐L fibreglass tanks (two tanks per genetic group) until they were 495 days old. At the end of the growth period fish were weighed, subjected to morphometric analysis and processed to obtain their bodily yields. Two nuclear markers and one mitochondrial marker were used to confirm the identity of the animals. Combining abilities were obtained using the method proposed by Griffing in 1956 (Australian Journal of Biological Science, 4, 463–493) adapted to a mixed models analysis, environmental effects were estimated by the empirical best linear unbiased estimator method and genetic effects (general and specific combining abilities) were estimated with the empirical best linear unbiased predictor. Predictions of the combining abilities of advanced hybrids were obtained by the mixed models mixture method with normal distributions. Tambaqui showed higher general and specific combining abilities than the other groups for most of the variables, making it the most important genetic group. General combining ability makes a greater contribution to phenotypic variance than specific combining ability for most variables, indicating a predominance of genes with an additive effect in the control of evaluated traits.

Made available in DSpace on 2018-12-11T17:17:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-02-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Tambaqui (Colossoma macropomum) is a fish species from the Amazon and Orinoco Rivers, with favorable characteristics to the cultivation system and great market acceptance in South America. However, the construction of a genetic map for the genetic improvement of this species is limited by the low number of molecular markers currently described. Thus, this study aimed to validate gene-associated and anonymous (non-genic) microsatellites obtained by next generation sequencing (RNA-seq and whole genome shotgun—WGS, respectively), for future construction of a genetic map and search for quantitative trait loci (QTL) in this species. In the RNA-seq data, the observed and expected heterozygosity (Ho and He) ranged from 0.09 to 0.73, and 0.09 to 0.85, respectively. In the WGS data, Ho and He ranged from 0.33 to 0.95, and 0.28 to 0.92, respectively. In general, the evaluation of 200 markers resulted in 45 polymorphic loci, of which 14 were gene-associated (RNA-Seq) and 31 were anonymous (WGS). Moreover, some markers were related to genes of the immune system, biological regulation/control and biogenesis. This study contributes to increase the number of molecular markers available for genetic studies in C. macropomum, which will allow the development of breeding programs assisted by molecular markers. Centro de Aquicultura da Unesp Universidade Estadual Paulista Departamento de Morfologia IBB Universidade Estadual Paulista Instituto do Mar Universidade Federal de São Paulo UNIFESP Departamento de Ciências Biológicas FC Universidade Estadual Paulista Centro de Aquicultura da Unesp Universidade Estadual Paulista Departamento de Morfologia IBB Universidade Estadual Paulista Departamento de Ciências Biológicas FC Universidade Estadual Paulista CNPq: 305916/2015-7 CNPq: CNPq 446779/2014-8 FAPESP: FAPESP 2014/03772-7

Made available in DSpace on 2018-11-26T17:48:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-02-22 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) PROPE/UNESP The pirapitinga, Piaractus brachypomus (Characiformes, Serrasalmidae), is a fish from the Amazon basin and is considered to be one of the main native species used in aquaculture production in South America. The objectives of this study were: (1) to perform liver transcriptome sequencing of pirapitinga through NGS and then validate a set of microsatellite markers for this species; and (2) to use polymorphic microsatellites for analysis of genetic variability in farmed stocks. The transcriptome sequencing was carried out through the Roche/454 technology, which resulted in 3,696 non-redundant contigs. Of this total, 2,568 contigs had similarity in the non-redundant (nr) protein database (Genbank) and 2,075 sequences were characterized in the categories of Gene Ontology (GO). After the validation process of 30 microsatellite loci, eight markers showed polymorphism. The analysis of these polymorphic markers in farmed stocks revealed that fish farms from North Brazil had a higher genetic diversity than fish farms from Southeast Brazil. AMOVA demonstrated that the highest proportion of variation was presented within the populations. However, when comparing different groups (1: Wild; 2: North fish farms; 3: Southeast fish farms), a considerable variation between the groups was observed. The F-ST values showed the occurrence of genetic structure among the broodstocks from different regions of Brazil. The transcriptome sequencing in pirapitinga provided important genetic resources for biological studies in this non-model species, and microsatellite data can be used as the framework for the genetic management of breeding stocks in Brazil, which might provide a basis for a genetic pre-breeding programme. Sao Paulo State Univ, Univ Estadual Paulista Julio Mesquita Filho, Aquaculture Ctr, Jaboticabal, Brazil Univ Santiago Compostela, Vet Fac, Lugo, Spain Sao Paulo State Univ, Sch Sci, Bauru, Brazil Sao Paulo State Univ, Univ Estadual Paulista Julio Mesquita Filho, Aquaculture Ctr, Jaboticabal, Brazil Sao Paulo State Univ, Sch Sci, Bauru, Brazil FAPESP: 2014/03772-7 FAPESP: 2014/05732-2 CNPq: 446779/2014-8 CNPq: 305916/2015-7

Made available in DSpace on 2018-12-11T17:17:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-01-18 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Tambaqui, Colossoma macropomum, is the main native fish species produced in Brazil, and is an important species for genetic improvement in aquaculture. In addition, breeding studies on this species can be optimized with the use of molecular markers associated with productive phenotypes. The objective of the present study was to test the performance of growth traits and resistance to the bacteria, Aeromonas hydrophila, in association with microsatellite markers in C. macropomum. In this study, three full-sib families were subjected to bacterial challenge and morphometric growth assessments. Tambaqui families subjected to the bacterial challenge differed significantly in death time and mortality rate. There was, however, no association between resistance to bacteria and microsatellite markers. In relation to growth traits, we observed a marker/phenotype association in two microsatellites. The marker in the 6b isoform x5 gene (TNCRC6b) was associated with length, whereas an anonymous marker was associated with height. The present study highlighted the evaluation of molecular markers associated with growth traits, and can serve as the basis for future marker-assisted selection (MAS) of tambaqui. Aquaculture Center of Unesp São Paulo State University (Unesp) School of Sciences São Paulo State University (Unesp) Aquaculture Center of Unesp São Paulo State University (Unesp) School of Sciences São Paulo State University (Unesp) CNPq: 130629/2015-4 FAPESP: 2014/03772-7 CNPq: 305916/2015-7 CNPq: 446779/2014-8

Made available in DSpace on 2018-11-26T17:40:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-11-01. Added 1 bitstream(s) on 2021-07-15T15:01:38Z : No. of bitstreams: 1 S0103-90162017000600443.pdf: 524774 bytes, checksum: 6cef05bd83bbdc4deac7ae9e9e4a8129 (MD5) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) This study aimed to impute the genetic makeup of individual fishes of Serrasalmidae family on the basis of body weight and morphometric measurements. Eighty-three juveniles, belonging to the genetic groups Pacu, Pirapitinga, Tambaqui, Tambacu, Tambatinga, Patinga, Paqui and Piraqui, were separated into 16 water tanks in a recirculation system, with two tanks per genetic group, where they remained until they reached 495 days of age. They were then weighed and analyzed according to the following morphometric parameters: Standard Length (SL), Head Length (HL), Body Height (BH), and Body Width (BW). The identity of each fish was confirmed with two SNPs and two mitochondrial markers. Two analyses were performed: one for the validating the imputation and another for imputing a genetic composition of animals considered to be advanced hybrids (post F1). In both analyses, we used linear mixed models with a mixture of normal distributions to impute the genetic makeup of the fish based on phenotype. We applied the mixed models method, whereby the environmental effects were estimated by the Empirical Best Linear Unbiased Estimator (EBLUE) and genetic effects are considered random, obtaining the Empirical Best Linear Unbiased Predictor (EBLUP) from the general (GCA) and the specific (SCA) combining ability effects. The results showed that validation of the genetic makeup imputation based on body weight can be used because of the strong correlation between the observed and imputed genotype. The fish classified as advanced hybrids had a genetic composition with a high probability of belonging to known genotypes and there was consistency in genotype imputation according to the different characteristics used. Goiano Fed Inst, Dept Anim Sci, Rod Goiana,Km 01, BR-75901970 Rio Verde, Go, Brazil Univ Fed Lavras, Dept Exact Sci, CP 3037, BR-37200000 Lavras, MG, Brazil Univ Fed Lavras, Dept Anim Sci, Lavras, MG, Brazil Univ Fed Lavras, Dept Food Sci, Lavras, MG, Brazil Sao Paulo State Univ, Sch Sci, Biol Sci Dept, Av Engn Luiz Edmundo Carrijo Coube 1401, BR-17033360 Bauru, SP, Brazil Sao Paulo State Univ, FCAV, Biol Sci Dept, Via Acesso Prof Paulo Donato Castellane S-N, BR-14884900 Jaboticabal, SP, Brazil Sao Paulo State Univ, Sch Sci, Biol Sci Dept, Av Engn Luiz Edmundo Carrijo Coube 1401, BR-17033360 Bauru, SP, Brazil Sao Paulo State Univ, FCAV, Biol Sci Dept, Via Acesso Prof Paulo Donato Castellane S-N, BR-14884900 Jaboticabal, SP, Brazil

B chromosomes have attracted the attention of Neotropical fish cytogeneticists in recent years, both for their remarkable occurrence in this group and also because of the interest in studies of the genetic structure and role played in the genome of these organisms. The aim of this study was to report the first occurrence of supernumerary chromosomes in Prochilodus argenteus (Agassiz, 1829), this being the fifth carrier species among thirteen within the genus Prochilodus (Agassiz, 1829). The extra elements identified in this species are small sized heterochromatic chromosomes characterized by a low mitotic instability index, being very similar to other supernumerary chromosomes described in the species of the genus Prochilodus. Morphology, structure and dispersion of the supernumerary genomic elements which occur in species of this genus are discussed aiming to better understand aspects involved the origin of supernumerary chromosomes and the differentiation process and relationships among species of this family.

The population of Prochilodus lineatus found in the Mogi-Guaçu River is karyotypically polymorphic, carrying acrocentric, metacentric, and submetacentric B chromosomes. The analysis of each B chromosome frequency in this species revealed a variation in the distribution pattern, with the metacentric type having the highest frequency (73.30%), followed by submetacentric (25.22%) and acrocentric B chromosomes (1.48%). The transmission pattern of the supernumerary chromosomes was identified by controlled crosses, and it was shown that the acro- and submetacentric B chromosomes have a transmission pattern below the Mendelian rate (kB = 0.333 and kB = 0.385, respectively), but the metacentric variant has a cumulative transmission pattern (kB = 0.587). These results indicate that the acro- and submetacentric B chromosomes are undergoing an extinction process, while the metacentric B chromosomes appear to be accumulating in frequency with each generation.

Made available in DSpace on 2018-12-11T17:06:44Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-03-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The presence of introgressive hybridization in the wild, especially that resulting from human interference, can have negative impacts on biodiversity. Genetic tools provide essential information for species and hybrid identification, facilitating the conservation of natural resources. Here, we tested a set of markers to precisely elucidate introgressive hybridization between Pseudoplatystoma corruscans and Pseudoplatystoma reticulatum, two South American catfishes. New microsatellites showed high interspecific genetic divergence, and simulated data demonstrated the high power of STRUCTURE and NEWHYBRIDS for hybrid identification and classification, especially when all 11 nuclear markers were used. The investigation of real populations suggested that natural hybridization is rare. Otherwise, different hybridization scenarios were observed in two wild populations: one involving advanced backcrosses and the other involving high admixture. Our data represent the first detailed evaluation of genetic introgression between these species in Parana and Paraguay Basins and suggest that genetic contamination is occurring through F1 hybrids from aquaculture facilities. The results also provide a useful set of markers for monitoring escapees to aid in the conservation of the wild population and sustainable aquaculture. Additionally, species genotypic data are freely available to be used in the future as “parental species reference” in Bayesian methods assignments. Universidade Estadual Paulista (UNESP) Geneaqua Centro de Aquicultura da Unesp (CAUNESP) Centro Nacional de Pesquisa e Conserva��o de Peixes Continentais Instituto Chico Mendes de Conserva��o (CEPTA/ICMBio) Universidade de Santiago de Compostela (USC) Departamento de Biologia Universidade Estadual Paulista (UNESP), Av. Eng. Luiz Edmundo Carrijo Coube 14-01, Vargem Limpa Universidade Estadual Paulista (UNESP) Centro de Aquicultura da Unesp (CAUNESP) Departamento de Biologia Universidade Estadual Paulista (UNESP), Av. Eng. Luiz Edmundo Carrijo Coube 14-01, Vargem Limpa CNPq: 477703/2011-9 FAPESP: 477703/2011-9

Made available in DSpace on 2022-04-29T08:26:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Within the genus Prochilodus Agassiz, 1829, five species are known to carry B chromosomes, i.e. chromosomes beyond the usual diploid number that have been traditionally considered as accessory for the genome. Chromosome microdissection and mapping of repetitive DNA sequences are effective tools to assess the DNA content and allow a better understanding about the origin and composition of these elements in an array of species. In this study, a novel characterization of B chromosomes in Prochilodus costatus Valenciennes, 1850 (2n=54) was reported for the first time and their sequence complementarity with the supernumerary chromosomes observed in Prochilodus lineatus (Valenciennes, 1836) and Prochilodus argenteus Agassiz, 1829 was investigated. The hybridization patterns obtained with chromosome painting using the micro B probe of P. costatus and the satDNA SATH1 mapping made it possible to assume homology of sequences between the B chromosomes of these congeneric species. Our results suggest that the origin of B chromosomes in the genus Prochilodus is a phylogenetically old event. Departamento de Biologia Animal Campus Universitário Universidade Federal de Viçosa Departamento de Morfologia Instituto de Biociências Universidade Estadual Paulista, Distrito de Rubião Junior, s/n Departamento de Ciências Biológicas Faculdade de Ciências Universidade Estadual Paulista, Campus de Bauru Departamento de Morfologia Instituto de Biociências Universidade Estadual Paulista, Distrito de Rubião Junior, s/n Departamento de Ciências Biológicas Faculdade de Ciências Universidade Estadual Paulista, Campus de Bauru

In contrast to the enormous variability of Neotropical fish species, genetic information about many groups is not yet available. Chromosomal studies have greatly contributed to the characterization of several species, providing important data on these animals. We performed the first cytogenetic characterization of the Amazonian fish species Brachyplatystoma filamentosum (Pimelodidae), popularly known as “piraiba”. The results revealed a diploid number of 2n = 56 composed of 24 metacentric, 12 submetacentric, and 10 acrocentric chromosomes in both females and males. After silver nitrate treatment (AgNORs) it was possible to visualize the nucleolus organizer region located in the terminal portion of the short arms of subtelocentric chromosome pair 22, presenting size polymorphism. Hybridization with 18S and 5S rDNA probes confirmed the number and location of 18S marks rDNA in pair 22 and differences in the size of signals among homologs. The 5S rDNA genes were localized near the centromere on the short arms...

Made available in DSpace on 2014-09-30T18:18:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-09-27Bitstream added on 2014-10-01T14:04:00Z : No. of bitstreams: 1 S0001-37652013000401371.pdf: 524136 bytes, checksum: c9655abf0df7d134b6893b2bf7e3a790 (MD5) Estudos citogenéticos foram desenvolvidos em Prochilodus lineatus (Valenciennes 1836), os quais descrevem um interessante sistema de pequenos cromossomos supranumerários. O objetivo deste trabalho foi estudar a frequência e morfologia dos cromossomos B em indivíduos a partir da linhagem parental e os padrões de herança destes elementos em indivíduos obtidos a partir de cruzamentos controlados na espécie P. lineatus. A taxa de transmissão dos cromossomos B revelaram um kB=0,388 para o tipo acrocêntrico, kB=0,507 para o tipo metacêntrico e kB=0,526 submetacêntrico. Os resultados obtidos levantam hipóteses de que os cromossomos B do tipo acrocêntrico se encontram em fase de extinção, enquanto que os supranumerários do tipo metacêntrico e submetacêntrico encontram-se em fase de neutralização, seguindo uma taxa de transmissão Mendeliana. Cytogenetic studies were developed in Prochilodus lineatus (Valenciennes 1836), describing an interesting system of small supernumerary chromosomes. The purpose of this work is to study the frequency and morphology of B chromosomes in individuals from the parental line and the inheritance patterns of these elements in individuals obtained from controlled crosses in the species P. lineatus. The transmission rate of B chromosomes revealed a kB=0.388 for the acrocentric type, a kB=0.507 for the metacentric type and a kB=0.526 for the submetacentric type. The obtained results raise hypothesis that B-acrocentric chromosomes are involved in an extinction process in this species, while the metacentric and submetacentric supernumerary elements comprises a neutral mechanism and follows a Mendelian transmission rate. Universidade Estadual Paulista Laboratorio de Genetica de Peixes Centro Nacional de Pesquisa e Conservacao de Peixes Continentais nstituto Chico Mendes de Conservacao da Biodiversidade Laboratorio de Genetica e Reproducao de Peixes Universidade Estadual Paulista Laboratorio de Biologia e Genetica de Peixes Universidade Estadual Paulista Laboratorio de Genetica de Peixes Universidade Estadual Paulista Laboratorio de Biologia e Genetica de Peixes

Departamento de Ciencias Biologicas Faculdade de Ciencias Universidade Estadual Paulista (UNESP), Bauru, SP

To ascertain the origin of B chromosomes in 2 fish species of the genus Prochilodus, i.e. P. lineatus and P. nigricans, we microdissected them and generated B-specific DNA probes. These probes were used to perform chromosome painting in both species and in 3 further ones belonging to the same genus (P. argenteus, P. brevis and P. costatus). Both probes hybridized with the B chromosomes in P. lineatus and P. nigricans, but with none of the chromosomes in the 5 species. This indicates that the B chromosomes have low similarity with DNAs located in the A chromosomes and suggests the possibility that the B chromosomes in the 2 species have a common origin. The most parsimonious explanation would imply intergeneric hybridization in an ancestor of P. lineatus and P. nigricans yielding the B chromosome as a byproduct, which remained in these 2 species after their phylogenetic origin, but was perhaps lost in other Prochilodus species. This hypothesis predicts that B chromosomes are old genomic elements in this genus, and this could be tested once a species from a relative genus would be found showing homology of its A chromosomes with the B-probes employed here, through a comparison of B chromosome DNA sequences with those in the A chromosomes of this other species.

A physical chromosome mapping of the H1 histone and 5S and 18S ribosomal RNA (rRNA) genes was performed in interspecific hybrids of Pseudoplatystoma corruscans and P. reticulatum. The results showed that 5S rRNA clusters were located in the terminal region of 2 chromosomes. H1 histone and 18S ribosomal genes were co-localized in the terminal portion of 2 chromosomes (distinct from the chromosomes bearing 5S clusters). These results represent the first report of association between H1 histone and 18S genes in fish genomes. The chromosome clustering of ribosomal and histone genes was already reported for different organisms and suggests a possible selective pressure for the maintenance of this association.

Made available in DSpace on 2018-12-11T17:03:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-06-01 Pacu (Piaractus mesopotamicus) is a Neotropical freshwater fish threatened by overfishing, and one of the species of highest commercial value for aquaculture. Genetic variability analysis through molecular markers is an essential technique to genetic management of the wild and cultivated stocks. The main objective of this study was to identify and validate gene-associated SNPs of the liver transcriptome of pacu. Through genetic analysis in one natural population (Paraná River), 32 polymorphic SNPs were successfully genotyped and validated, some of them related to immune system genes. The observed and expected heterozygosity ranged from 0.059 to 0.706 and 0.058 to 0.507, respectively. All loci were in Hardy–Weinberg equilibrium (P > 0.05). Our results showed useful genomic resources for pacu, with applicability in conservation purposes and aquaculture industry by using SNPs markers. Centro de Aquicultura da Unesp Universidade Estadual Paulista Departamento de Morfologia IBB Universidade Estadual Paulista Departamento de Genética Facultad de Veterinaria Universidad de Santiago de Compostela Departamento de Ciências Biológicas FC Universidade Estadual Paulista Centro de Aquicultura da Unesp Universidade Estadual Paulista Departamento de Morfologia IBB Universidade Estadual Paulista Departamento de Ciências Biológicas FC Universidade Estadual Paulista

Made available in DSpace on 2018-12-11T17:01:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-04-02 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Several cytogenetic studies in Prochilodus lineatus (Valenciennes 1836) describe a striking system of supernumerary microchromosomes revealing the occurrence of three morphological types visualized as acrocentric, metacentric and submetacentric. The present study aimed to compare the DNA homology among the B chromosome variants, as well as the components of the standard complement using total B specific probes obtained by microdissection. This technique, which allows direct isolation of DNA from each B chromosome variant of interest, was applied to obtain the probes. Accordingly, cross hybridizations targeting all types were performed by using the fluorescence in situ hybridization technique. The results obtained revealed signals of hybridization on all kinds of B chromosomes, indicating homology among the variants. Moreover, markings on the standard A complement were observed using the B submetacentric chromosome specific probe. Based on these data, we can infer that B chromosome variants in P. lineatus have homologous regions, suggesting a common origin from an ancestral variant. Furthermore, it can be also hypothesized that the primitive supernumerary was probably originated from elements of the standard complement, since the B submetacentric morphotype shares sequences with the centromeric region of some chromosomes of the A complement. Departamento de Ciências Biológicas Universidade Estadual Paulista (UNESP), Campus de Bauru Departamento de Morfologia Instituto de Biociências Universidade Estadual Paulista Departamento de Ciências Biológicas Universidade Estadual Paulista (UNESP), Campus de Bauru Departamento de Morfologia Instituto de Biociências Universidade Estadual Paulista

Individuals of Astyanax bockmanni from one population belonging to the Paranapanema River basin, southeastern Brazil, were cytogenetically examined. In the present paper, we report a diploid number of 50 chromosomes, with some individuals bearing additional chromosomes, characterized as different variants of B chromosomes. The B microchromosome of acrocentric morphology (Ba) was the most frequent type in this population (77.7%). Two other variants were identified as metacentric B chromosomes, but of distinct sizes, characterized as macrochromosome (BM) and microchromosome (Bm). Conspicuous heterochromatic blocks were observed in the terminal regions of some chromosomes of the standard karyotype and partially heterochromatic blocks were detected in all B chromosomes. 18S ribosomal RNA genes were distributed in the terminal region of several chromosomes, but were not located on the B chromosomes.

Molecular markers (Multiplex-PCR and PCR-RFLP of nuclear and mitochondrial genes) were used to assess the frequency of hybrids in wild stocks of two catfish species: Pseudoplatystoma corruscans and Pseudoplatystoma reticulatum . Analysis of fish from the Paraguay–Parana hydrographic basin (Brazil) revealed a low frequency of hybrids in the Parana River (3.60%), which might be the result of natural hybridisation. However, the frequency of hybrids was higher in the Mogi Guacu (50.00%) and Aquidauana (30.75%) Rivers, whose aquaculture systems support the majority of Brazil's fish farms, suggesting that the existence of hybrids in wild populations may be due to introduction or escapes from farm stocks. Furthermore, the identification of a post-F1 individual in the Mogi-Guacu River provides evidence of genetic introgression. These results show that safeguards are needed to ensure the correct handling of hybrids and the protection of native species.

Serrasalmid and pimelodid fish hybrids represent important advances for aqua- culture in Brazil, but they also constitute serious genetic risks to cultivated and natural populations. Serrasalmid hybrids ('tambacu', 'tambatinga' and 'patinga') result from crosses between Colossoma macropomum, Piaractus meso- potamicus and Piaractus brachypomus. Pimelodid hybrids ('ponto e virgula', 'cachandia´' and 'cachapira') arise from crosses between Pseudoplatystoma corruscans, Pseudoplatystoma reticulatum, Phractocephalus hemioliopterus and Leiarius marmoratus. The problems associated with hybrids mainly result from inappropriate use because these animals are reproductively compatible with their parental species. This review shows that monitoring of fish hybrids using genetic techniques is necessary for their sustainable development in aquacul- ture. The genetic technologies used to identify fish hybrids include cytogenetic methods, which are considered to be low cost, and polymerase chain reaction- based molecular markers, which are associated with high throughput. Therefore, both types of genetic methodologies should be applied in monitor- ing programs aimed at brood stock management, wild stocks, the trade of hybrid juveniles and processed fish products in markets. Moreover, physical and genetic confinement of hybrids in aquaculture operations will be necessary to avoid the problems posed by these animals. The expected result of these measures will be the production of genetically improved animals by fish farms, allowing the sector to develop further and offering high-quality animal protein.

Departamento de Ciencias Biologicas Faculdade de Ciencias Universidade Estadual Paulista (UNESP), Campus de Bauru, Bauru

Hybrids of fish obtained by crossing commercial species of the family Serrasalmidae (Pacu, Pirapitinga, and Tambaqui) have been produced on a large scale by the Brazilian aquaculture industry. However, because juvenile stages are morphologically indistinguishable, pure species and hybrid stocks may be easily mixed in breeding facilities, which may ultimately reduce production on farms and could have adverse impacts on native populations, through escape of fertile hybrids to the wild. We established molecular markers using mitochondrial and nuclear genes to identify hybrid lineages of Serrasalmids and characterize them in relation to their parental species, Colossoma macropomum, Piaractus mesopotamicus, and Piaractus brachypomus. Using these diagnostic molecular tools, we then examined fish purchased from two seed producers and detected substantial admixture of pure stocks and their various hybrids. Further application of these diagnostic genetic tools is needed to assess the impacts of hybridization and stock admixture, both in the aquaculture industry and in the environment.

We report here on the physical mapping of the H1 histone genes (hisDNA) and the 5S ribosomal DNA (rDNA) in 3 Neotropical fish species of the genus Astyanax(A. altiparanae, A. bockmanni and A. fasciatus) and the comparative analysis of the chromosomes bearing these genes. Nucleotide analyses by sequencing of both genes were also performed. The distribution of the H1 histone genes was more conserved than that of the rRNA genes, since these were always located in the pericentromeric regions of 2 chromosome pairs. 5S rDNA was found on one of the pairs that presented an H1 histone cluster; this seems to be a conserved chromosomal feature of the genus Astyanax. In addition, individuals of A. bockmanni and A. fasciatus showed clusters of 5S rDNA on 1 pair of acrocentric chromosomes, not found in A. altiparanae. The results obtained by chromosome mapping as well as by sequencing of both genes showed that A.bockmanni is more closely related to A. fasciatus than to A. altiparanae. The results allow the characterization of cytogenetic markers for improved elucidation of the processes involved in karyotype differentiation of fish genomes.

Prochilodus lineatus is widely used in pisciculture projects, mainly in the south of Brazil. It shows a basic karyotype composed of 2n = 54 chromosomes, in addition to as many as seven supernumerary chromosomes, also known as B-chromosomes. These additional small chromosomes are frequently heterochromatic, vary in number and morphology, and generally have no homology with the A complement. Intensive studies have investigated the function, origin and inheritance of these supernumerary chromosomes. The present study aimed to determine the inheritance pattern of the B-chromosomes resulting from directed cross-fertilization of P. lineatus, Prochilodontidae isolated from the Mogi-Guacu River, Pirassununga, SP. These cross-fertilization experiments were performed at CEPTA/ICMBio in Pirassununga. The transmission patterns of these micro-chromosomes agreed (KB= 0.48) with the expectation of regular meiotic behavior following a Mendelian transmission model (KB= 0.5). A non-accumulation process was observed for these B-chromosomes in filial generations. Together, our results indicate that the supernumerary chromosomes in P. lineatus samples from the Mogi-Guacu River exist in a neutral state and follow a Mendelian inheritance pattern.

The genus Prochilodus includes individuals ranging in size from medium to large, being highly relevant for commercial and subsistence fishing. Prochilodus species have a diploid number of 2n = 54 chromosomes and up to seven supernumerary (B) microchromosomes. Previous research has shown that B frequency increased drastically in the Mogi-Guaçu river population of Prochilodus lineatus in the early 1980s, whereas it remained about constant in the 1990s. Here we analyses B frequency in this population during the 2003-2007 period and have found that frequency has not changed significantly since 1987, and that these B chromosomes do not show the intra-individual variation in number that characterized them in the 1980s. This indicates that these B chromosomes have been neutralized, after their invasion, through their mitotic stabilization.

Within the genus Prochilodus, the species Prochilodus lineatus is certainly the most studied one from a cytogenetic point of view. In this study, a cytogenetic characterization of specimens of P. lineatus from Mogi-Guacu River was performed in the period from 2003 to 2007, through utilization of cytogenetic markers, such as Giemsa, Ag-NOR, C-banding and cytogenetical-molecular markers (FISH) to detect both 5S and 18S ribosomal genes. All analyzed individuals presented 2n=54 meta/submetacentric chromosomes, besides bearing up to 7 supernumerary microchromosomes. Polymorphic NORs were detected on a single chromosomal pair. The constitutive heterochromatin was distributed at centromeric region of all chromosomes in the A complement, while the microchromosomes were totally heterochromatic. A syntenic distribution of 5S and 18S ribosomal genes was detected, comprising the NOR-bearing chromosomal pair. No additional ribosomal clusters in other chromosomes were observed. Although the karyotype features are commonly conserved within the genus Prochilodus, the few differences on the distribution of both ribosomal genes and heterochromatin when compared to specimens of P. lineatus from other localities can be associated to the evolutionary changes that these repetitive sequences have undergone through the years.

Interspecific hybrids of fish can offer important advances in the zootechnical exploration of this animal group. However, hybrids can pose serious biological risks to the environment and natural populations, making genetic characterization and monitoring of hybrids produced in fish culture important concerns. In the present study, individuals of the hybrid ‘Piaupara’ were characterized by fluorescence in situ hybridization (FISH) using sex-specific probes to differentiate the respective parental contributions from female Leporinus macrocephalus (Piauçu) and male Leporinus elongatus (Piapara). The application of this technique demonstrated a difference in hybridization patterns between males and females of L. elongatus and L. macrocephalus, mainly due to the fact that the NOR-bearing chromosomes of L. elongatus contain some homology with the probe. This allowed us to detect a clear sex-specific distinction of the hybrid ‘Piaupara’, demonstrating that this technique is well-suited for the characterization of these hybrids. Moreover, this technique offers an important methodological advance for the study of diverse sex chromosome systems of Leporinus, supplying valuable information about the origin and establishment of a recently described sexual system in this genus.