Diagnostic locus for identification of silverside hybrids (Odontesthes humensis and Odontesthes bonariensis) in a natural population / Locus diagnóstico para identificação de híbridos de peixe-rei (Odontesthes humensis and Odontesthes bonariensis) em população natural

Hybridization is a natural phenomenon that occurs more often in fish than in other vertebrates. The use of nuclear and mitochondrial molecular markers provides valuable results in the detection of these events. The aim of this study was to investigate the occurrence of interspecific hybrids in natural populations of silverside. The samples of Odontesthes humensis, Odontesthes bonariensis, and indivi-duals that were morphologically different from pure species were collected in the Mangueira lagoon, located in southern Brazil. Result: Six tetranucleotide microsatellite loci were synthesized and tested. The UFPEL_OH3 locus proved to be diagnostic for the detection of silverside hybrids, and it was possi-ble to distinguish between pure and hybrid species. The mitochondrial marker gene cytb synthesized from conserved Odontesthes sequences in the GenBank genetic database showed no differences in the genetic sequence of the samples, needing further studies to confirm the hypothesis.(AU)

A hibridação é um fenômeno natural que ocorre mais frequentemente em peixes do que em outros vertebrados. Para detectá-la, são utilizados marcadores moleculares nucleares e mitocondriais, os quais fornecem resultados valiosos. O objetivo deste estudo foi investigar a ocorrência de híbridos in-terespecíficos em populações naturais de peixe-rei. As amostras de Odontesthes humensis, Odontesthes bonariensis e de indivíduos morfologicamente diferentes das espécies puras foram coletadas na lagoa Mangueira, localizada no Sul do Brasil. Foram sintetizados e testados seis loci microssatélites tetranu-cleotídeos. O locus UFPEL_OH3 mostrou-se um diagnóstico para detectar híbridos de peixe-rei, pos-sibilitando a distinção de espécies puras de híbridas. O marcador mitocondrial gene cytb, sintetizado com base nas sequências conservadas de Odontesthes no banco de dados genéticos do GenBank, não apresentou diferenças na sequência genética das amostras, portanto, são necessários mais estudos para confirmar a hipótese de hibridação.(AU)

Babesia bovis in large ruminants in Pakistan - molecular detection and haemato-biochemical alterations

Background: Babesiosis is endemic in Pakistan and is one of the most important bovine diseases that causes huge economic losses and high mortality in young animals. A hematobiochemical study was conducted to unveil the difference between diseased and healthy animals in selected districts i.e., Faisalabad (31° 25' 7.3740'' N and 73° 4' 44.7924'' E), Toba Tek Singh (30° 58' 9.7392'' N and 72° 27' 40.7484'' E) and Jhang (31° 16' 40.9656'' N and 72° 18' 42.3360'' E) of Punjab, Pakistan. Materials, Methods & Results: A total of 518 (Cattle = 360, Buffalo = 158) blood samples were collected. The samples were analyzed by polymerase chain reaction (PCR) targeting apocytochrome b-gene (Babesia bovis-gene) (CYTb) followed by haemato-biochemical analysis. Chi-square test for univariate analysis was used to analyze the data. In summer the PCR-based prevalence was 29.4 (53/180) and 24.05% (19/79) in cows and buffaloes, respectively. On the other hand, in winter results showed that 12.7 (23/180), 13.92 % (11/79) samples positive for Babesia genus from cows and buffaloes, respectively. The positive samples were further investigated for hematological and biochemical analysis. The results revealed that, the mean value of hematological parameters like RBCs, Hb, PCV, MCV and MCHC was significantly (P < 0.05) decreased in infected animals (cows and buffaloes) as compared to the non-infected ones. While the biochemical parameters like Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), cholesterol and Lactate dehydrogenase were significantly (P < 0.05) increased in infected animals as compared to healthy animals. This study is the first molecular and hematobiochemical evidence of Babesia bovis in dairy herds of Punjab province, Pakistan. Discussion: Bovine babesiosis is one of the important tick-borne diseases (TBD) affecting dairy industry. In bovines, among 3 Babesia species that cause the disease B. bovis is more pathogenic with high mortality and morbidity. Pakistan is situated in tropical and sub-tropical region where the humidity is high in some part of countries. This high humidity mostly favors the reproduction of the ticks thus higher prevalence of TBDs in this region. Initially the babesiosis was diagnosed by light microscopy using thin blood smear stained with Giemsa stain. Many studies verified that PCR is a more specific and sensitive tool than conventional techniques for the detection of carrier / asymptomatic ruminants. The haemato-biochemical profile is another valuable footprint to track the disease. Keeping in view the above-mentioned fact the present project has been planned to evaluate the haemato-biochemical alteration between health and Babesia infected cattle along with the molecular detection of Babesia species involved in bovine babesiosis. The mean values of haematobiochemical parameters in clinically ill and healthy animals revealed that the mean values of hematological parameters like RBCs, Hb, PCV, and HCT were significantly decreased in diseased animals as compared to the healthy ones. All these might be due the fact that the parasite is intra-erythrocytic in nature and destruction of red blood cells results in significant (P < 0.05) decrease level of all the hematological parameters. The mean value of ALT in babesiosis infected cattle was significantly higher as compared to healthy cattle. The mean values of AST and LDH in babesiosis infected cows was significantly higher as compared to that in healthy cows. The elevation in liver enzymes in babesiosis may be due to the hepatic damage and lesions induced by the parasite during its multiplication in the blood followed by disturbed liver function. These enzymes are present in high concentrations in the muscles and liver. High level of these enzymes in the blood is indicator of organ necrosis or damage.

Molecular epidemiology of bovine babesiosis in Punjab, Pakistan

Background: Babesiosis is endemic in Pakistan and is one of the most important bovine diseases that causes huge economic losses and high mortality in young animals. This disease is transmitted by a protozoan parasite, which belongs togenus Babesia (Apicomplexa: Piroplasmida: Babesiidae). This disease is very much prevalent in summers followed byrainy season because humid environment is favorable for the growth of these parasites. An epidemiological and molecularstudy was conducted to unveil the prevalence and associated risk factors of Babesia bigemina (B. bigemina) and Babesiabovis (B. bovis) in selected districts i.e., Faisalabad, Toba Tek Singh and Jhang of Punjab, Pakistan.Materials, Methods & Results: A total of 518 (Cattle = 360, Buffalo = 158) blood samples were collected. The sampleswere analyzed by polymerase chain reaction (PCR) and nested PCR (n-PCR) targeting apocytochrome b-genes (CYTb).Chi-square test for univariate analysis was used to analyze the data. The overall prevalence in summer based upon microscopic analysis was 20.55% (37/180) and 13.92% (11/79) in cattle and buffaloes respectively and in winter was 8.80%(16/180), 5.06% (4/79)) in cattle and buffaloes respectively. The samples were further analyzed through conventional PCR(c-PCR) and nested PCR (nPCR). The overall results of conventional PCR in summer showed that 72 cows and buffaloeswere infected with babesiosis. The conventional PCR based results of summer showed that prevalence of babesiosis was29.44% (53/180) in cows and 24.05% (19/79) buffaloes. The results of cPCR during the winter season showed that 12.77%(23/180) and 13.92% (11/79) buffaloes were positive for babesiosis. The overall results of conventional PCR in wintershowed that 34/259 cows and buffaloes were infected with babesiosis. On the other hand, the nested PCR results of summerseason showed that the prevalence of babesiosis in cows was 32.22% (58/180) and...

Molecular epidemiology of bovine babesiosis in Punjab, Pakistan

Background: Babesiosis is endemic in Pakistan and is one of the most important bovine diseases that causes huge economic losses and high mortality in young animals. This disease is transmitted by a protozoan parasite, which belongs togenus Babesia (Apicomplexa: Piroplasmida: Babesiidae). This disease is very much prevalent in summers followed byrainy season because humid environment is favorable for the growth of these parasites. An epidemiological and molecularstudy was conducted to unveil the prevalence and associated risk factors of Babesia bigemina (B. bigemina) and Babesiabovis (B. bovis) in selected districts i.e., Faisalabad, Toba Tek Singh and Jhang of Punjab, Pakistan.Materials, Methods & Results: A total of 518 (Cattle = 360, Buffalo = 158) blood samples were collected. The sampleswere analyzed by polymerase chain reaction (PCR) and nested PCR (n-PCR) targeting apocytochrome b-genes (CYTb).Chi-square test for univariate analysis was used to analyze the data. The overall prevalence in summer based upon microscopic analysis was 20.55% (37/180) and 13.92% (11/79) in cattle and buffaloes respectively and in winter was 8.80%(16/180), 5.06% (4/79)) in cattle and buffaloes respectively. The samples were further analyzed through conventional PCR(c-PCR) and nested PCR (nPCR). The overall results of conventional PCR in summer showed that 72 cows and buffaloeswere infected with babesiosis. The conventional PCR based results of summer showed that prevalence of babesiosis was29.44% (53/180) in cows and 24.05% (19/79) buffaloes. The results of cPCR during the winter season showed that 12.77%(23/180) and 13.92% (11/79) buffaloes were positive for babesiosis. The overall results of conventional PCR in wintershowed that 34/259 cows and buffaloes were infected with babesiosis. On the other hand, the nested PCR results of summerseason showed that the prevalence of babesiosis in cows was 32.22% (58/180) and...(AU)

Second capture of Promops centralis (Chiroptera) in French Guiana after 28 years of mist-netting and description of its echolocation and distress calls / Segunda captura de Promops centralis (Chiroptera) en la Guayana Francesa después de 28 años de muestreos con redes y descripción de su ecolocalización y sus llamadas de alarma

La cuenca amazónica alberga algunos de los ecosistemas más diversos en fauna quiropterológica del mundo. Sin embargo, en la literatura científica no encontramos información muy detallada sobre especies de murciélago esquivas como las de la família Molossidae. Esta carencia condiciona y obstaculiza los esfuerzos de conservación tanto a escala local como global. El desarrollo reciente de nuevas tecnologías diseñadas específicamente para muestrear quirópteros, como los detectores de ultrasonidos pasivos o los reclamos acústicos mediante el uso de llamadas de alta frecuencia, ha incrementado nuestro conocimiento sobre su ecología y distribución. Además, ha permitido a los investigadores obtener nuevos datos que eran prácticamente imposibles de conseguir en el pasado. Llevamos a cabo una evaluación rápida de diversidad quiropterológica en la Guayana Francesa, utilizando reclamos acústicos con el objetivo de capturar especies insectívoras de vuelo alto. En este estudio, aportamos la segunda y tercera captura de Promops centralis (Chiroptera, Molossidae) para Guayana Francesa después de 28 años desde sus primeras y únicas capturas hasta ahora. Uno de los indivíduos capturados fue una hembra poslactante, el primer registro de reproducción de la especie. Aportamos (i) datos morfométricos, bioacústicos (incluyendo las llamadas de alarma típicas de la especie) y fotografías de detalles para facilitar su identificación; y (ii) las secuencias de COI y CytB de los dos individuos (las primeras secuencias mitocondriales para la Guayana Francesa).(AU)

The Amazonian basin harbours some of the most bat-diverse ecosystems worldwide. Yet, information on elusive, high-flying bat species such as Molossidae is scarce or virtually missing in the literature, which hampers conservation efforts both locally and globally. The recent advent of new technologies specifically designed to survey bats, such as passive ultrasound detectors and acoustic lures, has significantly increased understanding of bat ecology and distribution, and has allowed researchers to gather new and valuable information which was impossible to collect in the past. We undertook a rapid bat diversity assessment in French Guiana using acoustic lures to aid in capturing high-flying insectivorous bat species. Here we report the second and third capture record of Promops centralis (Chiroptera, Molossidae) for French Guiana, captured after 28 years since the first and only captures so far in the county. One individual was a post-lactating female and represents the first record of breeding P. centralis in French Guiana. We provide (i) morphometric and acoustic data (including the species distress calls) as well as detail photography to aid in species identification; and (ii) COI and CytB sequences of the two individuals (first mitochondrial sequences for French Guiana).(AU)

Morphological and molecular characterization of Haemoproteus coatneyi and Haemoproteus erythrogravidus (Haemosporida: Haemoproteidae) in Passeriformes in Brazils Atlantic Forest / Caracterização morfológica e molecular de Haemoproteus coatneyi e Haemoproteus erythrogravidus (Haemosporida: Haemoproteidae) em Passeriformes da Mata Atlântica, Brasil

Haemoproteus spp. are protozoan parasites found in birds around the world. These parasites are identified through the morphology of gametocytes, phylogenetic analysis based on the mitochondrial cytb gene, and the parasites geographic distribution. The absence of erythrocytic merogony, high intraspecific genetic variation and low parasitemia in wild birds makes it essential to use integrative approaches that assist in the identification of these parasites. Thus, microscopic and molecular analyses, combined with spatial distribution, were carried out to verify the presence of Haemoproteus spp. in wild birds in Brazil. Light microscopy revealed one Tangara sayaca bird was parasitized by Haemoproteus coatneyi and, two specimens of Zonotrichia capensis presented Haemoproteus erythrogravidus. The morphology of the gametocytes of these two parasitic species showed high similarity. The molecular analysis revealed the presence of one lineage of H. coatneyi and two lineages of H. erythrogravidus, one of which is considered a new lineage. These lineages were grouped phylogenetically in separate clades, with low genetic divergence, and the H. erythrogravidus lineage emerged as an internal group of the lineages of H. coatneyi. The geographic distribution demonstrated that the two species occur in the American continent. This is the first report of H. erythrogravidus in Brazil.(AU)

Haemoproteus spp. são protozoários parasitos encontrados em aves de todo o mundo. A identificação desses parasitos é realizada por meio da morfologia dos gametócitos, da análise filogenética, baseada no gene mitoncodrial cytb e na distribuição geográfica do parasito. A ausência de merogonia eritrocítica, a alta variação genética intraespecífica e a baixa parasitemia em aves silvestres, tornam essencial a utilização de abordagens integrativas que auxiliem na identificação desses parasitos. Assim, análises microscópicas e moleculares, aliadas à distribuição espacial, foram realizadas para verificar a presença de Haemoproteus spp. em aves silvestres no Brasil. A microscopia óptica demonstrou que uma ave Tangara sayaca estava parasitada por Haemoproteus coatneyi, e dois espécimes de Zonotrichia capensis apresentavam Haemoproteus erythrogravidus, cujas morfologias dos gametócitos apresentaram alta similaridade. A análise molecular recuperou uma linhagem de H. coatneyi e duas linhagens de H. erythrogravidus, sendo uma dessas considerada nova linhagem. Essas linhagens se agruparam filogeneticamente em clados separados, apresentando baixa divergência genética, sendo que as linhagens de H. erythrogravidus emergiram como grupo interno às linhagens de H. coatneyi. A distribuição geográfica demonstrou que as duas espécies estão ocorrendo no continente americano. Este é o primeiro relato de H. erythrogravidus no Brasil.(AU)

Genetic analysis of whole mitochondrial genome of Lateolabrax maculatus (Perciformes: Moronidae) indicates the presence of two populations along the Chinese coast

The whole mitochondrial genome of Lateolabrax maculatus (Cuvier, 1828) was used to investigate the reasons for the observed patterns of genetic differentiation among 12 populations in northern and southern China. The haplotype diversity and nucleotide diversity of L. maculatus were 0.998 and 0.00169, respectively. Pairwise FST values between populations ranged from 0.001 to 0.429, correlating positively with geographic distance. Genetic structure analysis and haplotype network analysis indicated that these populations were split into two groups, in agreement with geographic segregation and environment. Tajimas D values, Fus Fs tests and Bayesian skyline plot (BSP) indicated that a demographic expansion event may have occurred in the history of L. maculatus. Through selection pressure analysis, we found evidence of significant negative selection at the ATP6, ND3, Cytb, COX3, COX2 and COX1 genes. In our hypotheses, this study implied that demographic events and selection of local environmental conditions, including temperature, are responsible for population divergence. These findings are a step forward toward the understanding of the genetic basis of differentiation and adaptation, as well as conservation of L. maculatus.(AU)

Genetic analysis of whole mitochondrial genome of Lateolabrax maculatus (Perciformes: Moronidae) indicates the presence of two populations along the Chinese coast

The whole mitochondrial genome of Lateolabrax maculatus (Cuvier, 1828) was used to investigate the reasons for the observed patterns of genetic differentiation among 12 populations in northern and southern China. The haplotype diversity and nucleotide diversity of L. maculatus were 0.998 and 0.00169, respectively. Pairwise FST values between populations ranged from 0.001 to 0.429, correlating positively with geographic distance. Genetic structure analysis and haplotype network analysis indicated that these populations were split into two groups, in agreement with geographic segregation and environment. Tajima’s D values, Fu’s Fs tests and Bayesian skyline plot (BSP) indicated that a demographic expansion event may have occurred in the history of L. maculatus. Through selection pressure analysis, we found evidence of significant negative selection at the ATP6, ND3, Cytb, COX3, COX2 and COX1 genes. In our hypotheses, this study implied that demographic events and selection of local environmental conditions, including temperature, are responsible for population divergence. These findings are a step forward toward the understanding of the genetic basis of differentiation and adaptation, as well as conservation of L. maculatus.

Does the widely distributed rodent Calomys tener (Cricetidae: Sigmodontinae) constitute a single evolutionary unit?

The nominal species Calomys tener (Winge, 1887) ranges broadly in open lands of the Caatinga, Cerrado, Pantanal and Mata Atlântica of Brazil, and was recently reported from the Pampas of southern Brazil, and in the Selva Paranaense of eastern Paraguay and northeastern Argentina. This rodent can be infected with the pathogenic Araraquara hantavirus in Brazil. Given that most epidemiological studies have not taken into account updated taxonomic findings of their rodent hosts, in this study, we obtained sequence data of the Cyt-b and COI genes of specimens of C. tener from 22 different geographical localities from throughout the currently known distribution of the species (including individuals from Argentina, Paraguay, Bolivia, and Brazil) to test if it constitutes a single genetic unit or if it presents genetic discontinuities that may represent different evolutionary lineages. Phylogenetic analyses including several species of Calomys recovered several clades with strong support. Regarding C. tener, it is recovered as sister to the node that cluster C. laucha (Fischer, 1814) sensu lato, C. expulsus (Lund, 1841) and species in the C. callosus (Rengger, 1830) species complex. At the intraspecific level there are no genetic gaps among haplotypes of C. tener that could suggest more than one species. The recent captures in the Pampas of southern Brazil and in the Selva Paranaense suggest that the species may be colonizing new geographic areas.

Does the widely distributed rodent Calomys tener (Cricetidae: Sigmodontinae) constitute a single evolutionary unit?

The nominal species Calomys tener (Winge, 1887) ranges broadly in open lands of the Caatinga, Cerrado, Pantanal and Mata Atlântica of Brazil, and was recently reported from the Pampas of southern Brazil, and in the Selva Paranaense of eastern Paraguay and northeastern Argentina. This rodent can be infected with the pathogenic Araraquara hantavirus in Brazil. Given that most epidemiological studies have not taken into account updated taxonomic findings of their rodent hosts, in this study, we obtained sequence data of the Cyt-b and COI genes of specimens of C. tener from 22 different geographical localities from throughout the currently known distribution of the species (including individuals from Argentina, Paraguay, Bolivia, and Brazil) to test if it constitutes a single genetic unit or if it presents genetic discontinuities that may represent different evolutionary lineages. Phylogenetic analyses including several species of Calomys recovered several clades with strong support. Regarding C. tener, it is recovered as sister to the node that cluster C. laucha (Fischer, 1814) sensu lato, C. expulsus (Lund, 1841) and species in the C. callosus (Rengger, 1830) species complex. At the intraspecific level there are no genetic gaps among haplotypes of C. tener that could suggest more than one species. The recent captures in the Pampas of southern Brazil and in the Selva Paranaense suggest that the species may be colonizing new geographic areas.(AU)

The genus Myotis (Chiroptera, Vespertilionidae) in Brazil: phylogeny, distribution, and cytogenetics / O gênero Myotis (Chiroptera, Vespertilionidae) no Brasil: filogenia, distribuição e citogenética

Myotis is the largest genus of the Vespertilionidae, showing a cosmopolitan geographical distribution and is considered an example of adaptive radiation. Nine species occurs in Brazil and this study synthesized aspects of the geographic distribution, karyotype, and phylogeny. A search in bibliographic databases was carried out using keywords. The phylogeny study was based on the sequencing of a specimen of Myotis ruber collected in a fragment of the Altantic Forest of Minas Gerais; this specimen was deposited at the Newton Baião de Azevedo Museum of Zoology. The genus showed to be widely distributed in the Brazilian territory, with Myotis nigricans being the most widespread. In addition, high karyotypic conservatism was observed in all species of the genus. The phylogenetic analyses using the mt-Cytb gene corroborated the monophyletic aspect of the genus and the Myotis ruber species.

Myotis é o maior gênero de Vespertilionidae, apresentando distribuição cosmopolita e sendo um excelente exemplo de radiação adaptativa. Nove espécies ocorrem no Brasil e este estudo sintetizou aspectos da distribuição geográfica, cariótipo e filogenia. Foi realizada uma pesquisa em bancos de dados bibliográficos, utilizando palavras-chave. O estudo de filogenia baseou-se no sequenciamento de um espécime de Myotis ruber, coletado em um fragmento de Mata Atlântica do Estado de Minas Gerais; e depositado no Museu de Zoologia Newton Baião de Azevedo. O gênero mostrou ser amplamente distribuído no território brasileiro, sendo Myotis nigricans a espécie mais representativa. Além disso, observou-se alto conservadorismo cariotípico em todas as espécies do gênero. As análises filogenéticas utilizando o gene mitocondrial citocromo-b corroboraram o aspecto monofilético do gênero e da espécie Myotis ruber.

The genus Myotis (Chiroptera, Vespertilionidae) in Brazil: phylogeny, distribution, and cytogenetics / O gênero Myotis (Chiroptera, Vespertilionidae) no Brasil: filogenia, distribuição e citogenética

Myotis is the largest genus of the Vespertilionidae, showing a cosmopolitan geographical distribution and is considered an example of adaptive radiation. Nine species occurs in Brazil and this study synthesized aspects of the geographic distribution, karyotype, and phylogeny. A search in bibliographic databases was carried out using keywords. The phylogeny study was based on the sequencing of a specimen of Myotis ruber collected in a fragment of the Altantic Forest of Minas Gerais; this specimen was deposited at the Newton Baião de Azevedo Museum of Zoology. The genus showed to be widely distributed in the Brazilian territory, with Myotis nigricans being the most widespread. In addition, high karyotypic conservatism was observed in all species of the genus. The phylogenetic analyses using the mt-Cytb gene corroborated the monophyletic aspect of the genus and the Myotis ruber species.(AU)

Myotis é o maior gênero de Vespertilionidae, apresentando distribuição cosmopolita e sendo um excelente exemplo de radiação adaptativa. Nove espécies ocorrem no Brasil e este estudo sintetizou aspectos da distribuição geográfica, cariótipo e filogenia. Foi realizada uma pesquisa em bancos de dados bibliográficos, utilizando palavras-chave. O estudo de filogenia baseou-se no sequenciamento de um espécime de Myotis ruber, coletado em um fragmento de Mata Atlântica do Estado de Minas Gerais; e depositado no Museu de Zoologia Newton Baião de Azevedo. O gênero mostrou ser amplamente distribuído no território brasileiro, sendo Myotis nigricans a espécie mais representativa. Além disso, observou-se alto conservadorismo cariotípico em todas as espécies do gênero. As análises filogenéticas utilizando o gene mitocondrial citocromo-b corroboraram o aspecto monofilético do gênero e da espécie Myotis ruber.(AU)

Molecular phylogeny of Toxoplasmatinae: comparison between inferences based on mitochondrial and apicoplast genetic sequences

Abstract Phylogenies within Toxoplasmatinae have been widely investigated with different molecular markers. Here, we studied molecular phylogenies of the Toxoplasmatinae subfamily based on apicoplast and mitochondrial genes. Partial sequences of apicoplast genes coding for caseinolytic protease (clpC) and beta subunit of RNA polymerase (rpoB), and mitochondrial gene coding for cytochrome B (cytB) were analyzed. Laboratory-adapted strains of the closely related parasites Sarcocystis falcatula and Sarcocystis neurona were investigated, along with Neospora caninum, Neospora hughesi, Toxoplasma gondii (strains RH, CTG and PTG), Besnoitia akodoni, Hammondia hammondiand two genetically divergent lineages of Hammondia heydorni. The molecular analysis based on organellar genes did not clearly differentiate between N. caninum and N. hughesi, but the two lineages of H. heydorni were confirmed. Slight differences between the strains of S. falcatula and S. neurona were encountered in all markers. In conclusion, congruent phylogenies were inferred from the three different genes and they might be used for screening undescribed sarcocystid parasites in order to ascertain their phylogenetic relationships with organisms of the family Sarcocystidae. The evolutionary studies based on organelar genes confirm that the genusHammondia is paraphyletic. The primers used for amplification of clpC and rpoB were able to amplify genetic sequences of organisms of the genus Sarcocystisand organisms of the subfamily Toxoplasmatinae as well.

Resumo A filogenia da subfamília Toxoplasmatinae tem sido amplamente investigada com diversos marcadores moleculares. Neste estudo, a filogenia molecular da subfamília Toxoplasmatinae foi analisada através de genes de apicoplasto e mitocondriais. Foram analisadas sequências parciais de genes de apicoplasto codificadores da protease caseinolítica (clpC), e da subunidade beta da RNA polimerase (rpoB) e de gene mitocondrial codificador de citocromo B (cytB). Foram investigadas cepas adaptadas em laboratório de Sarcocystis neurona eSarcocystis falcatula, parasitos estreitamente relacionados, além de Neospora caninum, Neospora hughesi, Toxoplasma gondii (cepas RH, CTG e PTG),Besnoitia akodoni, Hammondia hammondi e duas linhagens geneticamente divergentes de Hammondia heydorni. A análise molecular, baseada em genes de organelas, não diferenciou claramenteN. caninum de N. hughesi, porém foi possível confirmar as duas linhagens de H. heydorni. Foram encontradas pequenas diferenças entre as cepas adaptadas em laboratório deS. falcatula e S. neurona em todos os marcadores moleculares avaliados. Concluindo, filogenias congruentes foram reconstruídas com os três diferentes genes que podem ser úteis em triagem de parasitos sarcocistídeos não identificados, para identificar sua relação com organismos da família Sarcocystidae. Os estudos evolutivos com genes organelares confirmam que o gênero Hammondia é parafilético. Osprimers utilizados para amplificação declpC e rpoB foram capazes de amplificar sequências genéticas de organismos do gênero Sarcocystis e da subfamília Toxoplasmatinae.

Infecção por piroplasmídeos em cães e gatos no Distrito Federal

Piroplasmídeos são hemoprotozoários pertencentes ao Filo Apicomplexa. Em cães e gatos, os gêneros mais importantes são Babesia spp. e Theileria spp., além de Cytauxzoon spp. e Rangelia vitalii somente em felinos e caninos, respectivamente. No Brasil, apesar de já terem sido descritos casos desses piroplasmídeos em cães e gatos, a caracterização clínica, laboratorial e molecular é incipiente. Com o objetivo de investigar a ocorrência, bem como de realizar a caracterização clínico-patológica dessas infecções, o presente estudo analisou amostras de sangue total de 276 cães e 171 gatos domésticos oriundos de Brasília, centro-oeste do Brasil. As amostras foram submetidas à Reação em Cadeia pela Polimerase quantitativa (qPCR) direcionada ao gene mitocondrial LSU4. A ocorrência em cães foi de 9,7% (27/276) para Babesia vogeli e 1,44% (4/276) para Piroplasma sp. não identificado. Com relação aos felinos, foi observada a maior ocorrência de Cytauxzoon já descrita no país, de 7% (12/171), e 11,11% (19/171) para Babesia vogeli. Não foi detectada nenhuma amostra positiva para Theileria spp., tanto para cães quanto para gatos, tampouco para Rangelia vitalii. Em cães infectados foi observado risco 2,3 vezes maior de apresentar trombocitopenia, além de uma tendência a anemia, em comparação aos animais não infectados. Não houve diferença significativa na análise hematológica e bioquímica dos felinos. Todas as amostras positivas foram submetidas posteriormente à caracterização molecular por meio de PCR convencionais (cPCR) com o gene 18S rRNA e os marcadores mitocondriais cox-1 e Cytb. Em cães, a análise filogenética destacou a espécie Babesia vogeli como a mais prevalente em cães do Brasil, além de posicionar em clado distinto Piroplasmas não identificados. A rede de haplótipos realizada com sequências cox-1 dos cães e gatos positivos revelou a existência de seis haplótipos para Babesia vogeli, sendo que um mesmo haplótipo é compartilhado com isolados do Brasil, EUA e Índia. Nossos estudos sugerem a existência de um piroplasmídeo distinto de B. vogeli que infecta cães da região. Quanto aos felinos, a infecção por Babesia vogeli e Cytauxzoon sp. aparentemente não ocasiona alterações clínicas. Uma baixa variabilidade intraespecífica foi observada nas sequências cox-1 de Babesia vogeli.

Piroplasmida are hemoprotozoans that belong to Apicomplexa phylumIn dogs and cats, they encompass the genera Babesia spp and Theileria spp, besides Cytauxzoon spp. and Rangelia vitalii only for felines and canids, respectively. In Brazil, although there are some cases of these piroplasms in dogs and cats, the clinical, laboratory and molecular characterization is incipient. To investigate the occurrence as well as the clinicopathological characterization of these infections, the current study analyzed total blood samples of 276 dogs and 171 domestic cats from Brasilia, Midwestern Brazil. All samples were carried out with the Quantitative Polymerase Chain Reaction (qPCR) using the mitochondrial gene LSU. The occurrence in dogs for Babesia vogeli was 9.7% (27/276), and 1.44% (4/276) for a non-identified Piroplasm sp. Regarding the felines, it has been observed the highest occurrence of Cytauxzoon sp. ever found in the country (7%; 12/171), and 11.11% (19/171) for Babesia vogeli. None of the samples were positive for Theileria spp., for both dogs and cats, neither for Rangelia vitalii. Infected dogs were 2.3 times at higher risk of having thrombocytopenia, in addition to a tendency of anemia, compared with the non-infected dogs. There was no significant difference concerning the hematologic and biochemistry analysis of cats. Subsequently, all positive samples were submitted to a conventional PCR characterization, using the 18S rRNA and the molecular markers cox-1 and Cytb. For dogs, the phylogenetic analysis enhanced Babesia vogeli species as the most prevalent in Brazil, in addition to positioning the Piroplasm sp. in a distinct clade. The haplotype network was performed using the cox-1 sequences from dogs and cats, which found six haplotypes for Babesia vogeli, one of them is shared with isolates from Brazil, the USA and India. Our studies suggest the presence of a piroplasm different from Babesia vogeli species, that infects dogs from the region. Regarding the felines, the infection by Babesia vogeli and Cytauxzoon sp. apparently did not show clinical relevance. A low intraspecific variability regarding B. vogeli cox-1 gene sequences was found.

Molecular phylogeny of Toxoplasmatinae: comparison between inferences based on mitochondrial and apicoplast genetic sequences / Filogenia molecular de Toxoplasmatinae: comparação entre inferências baseadas em sequências genéticas mitocondriais e de apicoplasto

Phylogenies within Toxoplasmatinae have been widely investigated with different molecular markers. Here, we studied molecular phylogenies of the Toxoplasmatinae subfamily based on apicoplast and mitochondrial genes. Partial sequences of apicoplast genes coding for caseinolytic protease (clpC) and beta subunit of RNA polymerase (rpoB), and mitochondrial gene coding for cytochrome B (cytB) were analyzed. Laboratory-adapted strains of the closely related parasites Sarcocystis falcatula and Sarcocystis neurona were investigated, along with Neospora caninum, Neospora hughesi, Toxoplasma gondii (strains RH, CTG and PTG), Besnoitia akodoni, Hammondia hammondiand two genetically divergent lineages of Hammondia heydorni. The molecular analysis based on organellar genes did not clearly differentiate between N. caninum and N. hughesi, but the two lineages of H. heydorni were confirmed. Slight differences between the strains of S. falcatula and S. neurona were encountered in all markers. In conclusion, congruent phylogenies were inferred from the three different genes and they might be used for screening undescribed sarcocystid parasites in order to ascertain their phylogenetic relationships with organisms of the family Sarcocystidae. The evolutionary studies based on organelar genes confirm that the genusHammondia is paraphyletic. The primers used for amplification of clpC and rpoB were able to amplify genetic sequences of organisms of the genus Sarcocystisand organisms of the subfamily Toxoplasmatinae as well.(AU)

A filogenia da subfamília Toxoplasmatinae tem sido amplamente investigada com diversos marcadores moleculares. Neste estudo, a filogenia molecular da subfamília Toxoplasmatinae foi analisada através de genes de apicoplasto e mitocondriais. Foram analisadas sequências parciais de genes de apicoplasto codificadores da protease caseinolítica (clpC), e da subunidade beta da RNA polimerase (rpoB) e de gene mitocondrial codificador de citocromo B (cytB). Foram investigadas cepas adaptadas em laboratório de Sarcocystis neurona eSarcocystis falcatula, parasitos estreitamente relacionados, além de Neospora caninum, Neospora hughesi, Toxoplasma gondii (cepas RH, CTG e PTG),Besnoitia akodoni, Hammondia hammondi e duas linhagens geneticamente divergentes de Hammondia heydorni. A análise molecular, baseada em genes de organelas, não diferenciou claramenteN. caninum de N. hughesi, porém foi possível confirmar as duas linhagens de H. heydorni. Foram encontradas pequenas diferenças entre as cepas adaptadas em laboratório deS. falcatula e S. neurona em todos os marcadores moleculares avaliados. Concluindo, filogenias congruentes foram reconstruídas com os três diferentes genes que podem ser úteis em triagem de parasitos sarcocistídeos não identificados, para identificar sua relação com organismos da família Sarcocystidae. Os estudos evolutivos com genes organelares confirmam que o gênero Hammondia é parafilético. Osprimers utilizados para amplificação declpC e rpoB foram capazes de amplificar sequências genéticas de organismos do gênero Sarcocystis e da subfamília Toxoplasmatinae.(AU)

Prospecting molecular markers to distinguish Cichla kelberi, C. monoculus and C. piquiti / Prospecção de marcadores moleculares para distinção de Cichla kelberi, Cichla monoculus e Cichla piquiti

Peacock bass, a fish of the genus Cichla, is an exotic species from the upper river Paraná floodplain in which the species Cichla kelberi and C. piquiti have been confirmed, coupled to the specie C. monoculus upstream in the Capivara and Taquaruçu dams. The introduction of this genus has caused negative impacts on the diversity of native species. Current research prospects DNA sequences capable of distinguishing the three species and provide molecular data for the taxonomic characterization of the species in the upper Paraná River basin.  Sequencing of nuclear (tmo4c4, dlx2 and bmp4) and mitochondrial (cox1, cytb) loci were done from fish of the three species of the genus Cichla reported in the literature of the upper Paraná River basin. Sequence analysis provided molecular differentiation for the species through the usage of loci cytb, dlx2 and cox1. Since the latter only distinguished C. piquiti from the other Cichla species, the loci bmp4 and tmo4c4 were not adequate to accomplish our aim.(AU)

O tucunaré é um peixe pertencente ao gênero Cichla, sendo exótico na planície de inundação do alto rio Paraná, onde foi confirmada a presença das espécies Cichla kelberi e C. piquiti e, logo a montante, C. monoculus, nas represas de Capivara e Taquaruçu. A introdução deste gênero tem ocasionado impacto negativo à diversidade de espécies nativas. Visando providenciar dados moleculares para auxiliar na caracterização taxonômica das espécies deste gênero na bacia do alto rio Paraná, os objetivos deste trabalho foram prospectar sequências de DNA capazes de distinguir as três espécies. Procedeu-se o sequenciamento de loci nucleares (tmo4c4, dlx2 e bmp4) e mitocondriais (cox1, cytb) de peixes das três espécies do gênero Cichla relatadas na literatura da bacia do alto rio Paraná. As análises das sequências possibilitaram a diferenciação molecular para estas espécies pela utilização dos loci cytb, dlx2 e cox1; este último, somente para distinguir C. piquiti das demais espécies de Cichla. A utilização dos loci bmp4 e tmo4c4 não foi adequada para este propósito.(AU)

Estimativa da diversidade genética do peixe-rei (Odontesthes humensis e Odontesthes bonariensis) por marcadores de microssatélite e CYTB nas lagoas Mirim e Mangueira no Rio Grande do Sul

Apesar da diversidade de peixes e da sua grande importância econômica para o homem, ainda pouco se conhece sobre suas características biológicas, ecológicas e genéticas. A falta de conhecimento das espécies e consequentemente a falta de manejo destas, além das altas taxas de pesca são os principais responsáveis da perda de diversidade e reduções drásticas nas populações. Este trabalho teve como objetivo, caracterizar geneticamente populações de peixe-rei, de duas lagoas, através de análises genéticas de polimorfismo através marcadores microssatélites e mitocondriais. A primeira parte do estudo foi realizada em duas lagoas, Mirim e Mangueira, localizadas no sul do Brasil de onde foram obtidas amostras de Odontesthes humensis e Odontesthes bonariensis. Foram sintetizados primers para a amplificação de seis loci microssatélites os quais foram validados e apresentaram alto polimorfismo nas populações. Foram obtidos no total 38 alelos e apenas uma população está em Equilíbrio de Hardy-Weinberg. Na segunda parte do estudo foram coletadas amostras de peixe-rei na lagoa Mangueira, que morfologicamente eram diferentes das espécies puras. O locus UFPEL_OH3 se mostrou um primer diagnóstico para a detecção de híbridos de peixe-rei. Foi sintetizado um conjunto de primers para amplificação de um fragmento do gene mitocondrial cytb para para confrontar as sequências mitocondriais das amostras em estudo e com sequências preservadas de Odontesthes do banco genético GenBank. O estudo sugere que possam estar ocorrendo eventos de especiação no ambiente, talvez associados a mudanças ambientais e efeito de sobrepesca nas lagoas em estudo, sendo necessária a elaboração de projetos de conservação e manejo da espécie.

Despite the diversity of fish and their great economic importance for man, little is known about their biological, ecological and genetic characteristics. The lack of knowledge of the species and consequently the lack of management of these, in addition to the high rates of fishing are the main responsible for the loss of diversity and drastic reductions in the populations. The objective of this work was to genetically characterize populations of silverside, from two lagoons, through genetic analyzes of polymorphism using microsatellite and mitochondrial markers. The first part of the study was carried out in two lagoons, Mirim and Mangueira, located in southern Brazil from where samples of Odontesthes humensis and Odontesthes bonariensis were obtained. Six microsatellite loci were synthesized which were validated and showed high polymorphism in the populations. A total of 38 alleles were obtained and only one population is in Hardy-Weinberg equilibrium. In the second part of the study, samples of silverside were collected in the Mangueira lagoon, which were morphologically different from pure species. The UFPEL_OH3 locus proved to be a diagnostic primer for the detection of silverside hybrids. Mitochondrial marker (cyt b) was synthesized to compare the mitochondrial sequences of the samples under study and with preserved Odontesthes sequences from the GenBank genetic bank. The study suggests that speciation events may be occurring in the environment, perhaps associated with environmental changes and the effect of overfishing in the ponds under study, requiring the elaboration of conservation and management projects for the species.

FILOGENIA E DELIMITAÇÃO MOLECULAR DE ESPÉCIES DO GÊNERO Mazama RAFINESQUE, 1817 (CETARTIODACTYLA:CERVIDAE)

O veado-mateiro (Mazama americana Erxleben, 1777) representa um complexo de espécies crípticas cujo caráter polifilético e diferenças cromossômicas já foram demonstrados. Dependendo do grau de divergência, essas diferenças são uma barreira reprodutiva que levam ao isolamento e reforçam a necessidade de revisão taxonômica do grupo. Assim, foi proposta a revalidação da espécie Mazama rufa Illiger 1811 para Mata Atlântica. A resolução da incerteza taxonômica que ainda cerca o complexo é vital na avaliação e conservação do grupo, posto que várias lacunas amostrais e espécies em potencial carecem de avalição. Nesse sentido, o presente trabalho buscou avaliar as relações filogenéticas entre as variantes cromossômicas do complexo M. americana e delimitar potenciais novas espécies a partir de dados moleculares. Também foi testada a validade da espécie M. rufa e inferidos aspectos de sua distribuição e estado de conservação. Um conjunto de espécimes com informação cromossômica (citótipo) e outro, de amostras fecais coletadas na natureza, foram sequenciados para parte dos genes mitocondriais Cytb, D-loop e ND5. Foram realizadas análises filogenéticas por meio de inferência Bayesiana e Máxima Verossimilhança e aplicados métodos coalescentes de delimitação molecular de espécie (GMYC e PTP) para identificar unidades taxonômicas operacionais moleculares (MOTUs). A hipótese filogenética mostrou relação de monofilia recíproca entre todos os citótipos, com exceção do citótipo Carajás. As espécies M. americana (sensu strictu) e M. rufa foram consideradas como espécies irmãs e válidas. Já os citótipos Juína e Rondônia representam duas potenciais espécies distintas de M. americana. A espécie M. rufa é composta pelos citótipos Paraná e Carajás, apresenta ampla distribuição, do sul do Brasil até a faixa sul-sudeste da Amazônia, e potencialmente não deve ser classificada como ameaçada de extinção. Ficou demonstrado que até a menor diferença cromossômica (uma fusão em tandem) pode implicar em espécies distintas, dependendo do tempo de divergência.

The red brocket deer (Mazama americana Erxleben, 1777)) is considered cryptic species complex whose polyphyletic state and chromosomal differences have been already demonstrated. These chromossomal differences act as a reproductive barrier that can lead to isolation, depending on the level of divergence, thus, reinforce the need for taxonomic revision. In this sense a species revalidation has just been proposed for Mazama rufa Illiger 1811 in the Atlantic Forest. The resolution of the remaning taxonomic uncertainty is vital in this taxa assessment and conservation, once there is several sampling gaps and other potential species to be evaluated. Therefore, the present work sought to evaluate the phylogenetic relationships between the chromosomal variants and to delimit potential new species for the M. americana complex from molecular data. The validity of M. rufa was tested and aspects of its distribution and conservation status were also inferred. A set of specimens with chromosomal information (cytotype) and another set of fecal samples collected in nature were sequenced for part of the mitochondrial regions Cytb, D-loop and ND5. Bayesian inference and Maximum Likelihood phylogenetic analyzes were performed and coalescent methods of molecular species delimitation (GMYC and PTP) were applied to identify molecular operational taxonomic units (MOTUs). The phylogenetic hypothesis demonstrated a reciprocal monophiletic relationship between all cytotypes, with the exception of Carajás. The red brocket deers M. americana (sensu strictu) and M. rufa were recovered as sisters and valid species. On the other hand, Juína and Rondônia cytotypes should represent two distinct species from M. americana. The M. rufa species is composed of Paraná and Carajás cytotypes, has a wide distribution from the south of Brazil up to the south-southeast of the Amazon Forest, and potentially will not be categorized as threatened with extinction. The results demonstrated that even the minimal chromosomal difference (a tandem fusion) can imply different species, depending on other factors as the time since divergence.

Análise transcricional do desenvolvimento pré-natal de suínos

A trajetória do desenvolvimento dos conceptos (embriões e fetos) é caracterizada como um evento biológico altamente regulado a nível transcricional, podendo ser influenciada pelo dimorfismo sexual e nutrição materna. O dimorfismo sexual no início do desenvolvimento pré-natal, período no qual a diferenciação sexual com base no fenótipo é dificultada, está principalmente associado à expressão de genes ligados aos cromossomos sexuais. A nutrição materna é o principal fator extrínseco que afeta a programação do desenvolvimento, dada a influência que as moléculas resultantes do metabolismo dietético exercem sobre o ambiente intrauterino, as quais são importantes não apenas por proporcionar nutrientes para o crescimento dos conceptos, mas também por regular a transcrição de genes relacionados ao desenvolvimento da placenta e dos conceptos. Neste contexto, os efeitos da suplementação de fêmeas suínas gestantes com aminoácidos funcionais como a arginina sob aspectos produtivos e reprodutivos têm sido bem estudados. Entretanto, a influência dessa suplementação sobre a expressão gênica global durante o desenvolvimento dos conceptos ainda é pouco conhecida. O adequado desenvolvimento e funcionalidade de órgãos e tecidos são eventos determinados principalmente durante a embriogênese e o período fetal, e requerem um ajustado controle da expressão gênica. Embora em espécies acometidas por restrição do crescimento intrauterino, como os suínos, o conhecimento da expressão de genes relacionados a organogênese possa auxiliar no melhor entendimento de vários aspectos da biologia do desenvolvimento pré-natal este ainda tem sido pouco explorado. Atualmente, o conhecimento da expressão gênica bem como dos processos biológicos e vias envolvidas na regulação gênica tem sido possibilitada pela análise do transcriptoma por meio do sequenciamento de RNA (RNA-Seq) e análises funcionais. Dessa forma, no presente trabalho, a partir do RNA-seq de embriões aos 25 dias de gestação e fetos suínos aos 35 dias de gestação, nós objetivamos (1) determinar o sexo desses conceptos em estágios iniciais do desenvolvimento; (2) elucidar os efeitos da suplementação de marrãs gestantes com L-arginina sobre o transcriptoma dos conceptos; e, (3) evidenciar os principais mecanismos moleculares envolvidos na organogênese em suínos, por meio da identificação de genes diferencialmente expressos entre embriões e fetos. Os resultados desta tese mostraram que os genes ligados ao cromossomo Y (DDX3Y, KDM5D, ZFY, EIF2S3Y, EIF1AY, LOC110255320, LOC110257894, LOC396706, LOC100625207 e LOC110255257) apresentaram contagens discrepantes entre as amostras de embriões e fetos suínos as quais foram utilizadas para a determinação do sexo dos conceptos. A suplementação dietética de marrãs com 1% de L-arginina provocou redução na expressão de genes mitocondriais (ND1, ND2, CYTB, COX2, ATP8, e mt-rRNA type) e aumento na expressão do gene CYP1A1 em embriões, evidenciando um ajustado controle transcricional a nível metabólico, enquanto não provocou alterações transcricionais durante o desenvolvimento fetal. Durante a organogênese em suínos, 1705 genes foram diferencialmente expressos entre embriões e fetos, a partir dos quais várias vias e processos biológicos relacionados à organogênese renal, muscular esquelética e óssea, bem como o desenvolvimento dos sistemas cardiovascular e neuronal, foram evidenciados. Esta tese proporcionou relevantes informações sobre o desenvolvimento pré-natal de suínos que podem ser aplicados em estudos relacionados à eficiência na produção de suínos, bem como no maior entendimento da biologia do desenvolvimento.

The conceptuses (embryos and fetuses) developmental trajectory is characterized as a highly regulated biological event at the transcriptional level, which may be influenced by sexual dimorphism and maternal nutrition. Sexual dimorphism at early developmental period, in which the phenotypic sex differentiation is not evident, is mainly related to the expression of genes linked to sex chromosomes. Maternal nutrition is the main extrinsic factor that affects developmental programming, since the molecules from dietary metabolism affects the intrauterine environment, which are important not only to provide nutrients to the conceptuses growth, but also for transcriptional regulation of genes related to placental and conceptuses development. In this context, the effects of pregnant females supplementation with functional amino acids, such as arginine, have been well studied under productive and reproductive aspects in pig production. However, the influence of this supplementation on global gene expression during conceptuses development is still poorly understood. The proper development and functionality of organs and tissues are events determined during embryogenesis and the fetal period, which require an adjusted control of gene expression. Although in species affected by intrauterine growth restriction, such as pigs, the knowledge about genes expression related to organogenesis can help to understand various aspects associated with prenatal development, studies addressing this event are still scarce. Currently, the gene expression, as well as the biological processes and pathways involved in its regulation, has been performed by transcriptome analysis from RNA sequencing (RNA-Seq) and functional analyses. Thus, in the present study, from the RNA-seq of pig embryos at 25 days of gestation and fetuses at 35 days of gestation, we aimed (1) the conceptuses sex determination; (2) to elucidate the effects of pregnant gilts supplemented with L-arginine on the conceptuses transcriptome; and, (3) to reveal the main molecular mechanisms involved in pig organogenesis, through the identification of genes differentially expressed between embryos and fetuses. The results of this thesis showed that Y chromosome-linked genes (DDX3Y, KDM5D, ZFY, EIF2S3Y, EIF1AY, LOC110255320, LOC110257894, LOC396706, LOC100625207 and LOC110255257) presented discrepant reads counts between the embryos and fetuses samples, which were used for conceptuses sex determination at early prenatal development. Dietary 1.0% L-arginine supplementation of pregnant gilts resulted in downregulation of mitochondrial genes (ND1, ND2, CYTB, COX2, ATP8, and mt-rRNA type) and upregulation of the CYP1A1 gene in embryos, showing an adjusted transcriptional control at the metabolic level, while the transcriptional program during fetal development was not affected. During the pig organogenesis, 1,705 genes were differentially expressed between embryos and fetuses, from which several biological pathways and processes related to renal, skeletal and bone muscular organogenesis, as well as the development of the cardiovascular and neuronal systems were evidenced. This thesis provided relevant information about the pig prenatal development, which can be applied in studies related to pig production efficiency, as well as greater understanding of developmental biology.

HISTÓRIA DE VIDA DE Aetobatus narinari (MYLIOBATIFORMES) CAPTURADA PELA PESCA ARTESANAL NO LITORAL DA PARAÍBA E PERNAMBUCO, BRASIL

A raia pintada, Aetobatus narinari, ocorre em regiões tropicais costeiras e insulares no oceano Atlântico. A recente redefinição taxonômica da espécie reduziu a literatura disponível sobre sua biologia e a pesca, bem como tornou seu status de conservação (Quase Ameaçada - NT) obsoleto a nível mundial. No Brasil, essa raia é capturada pela pesca artesanal em várias regiões, mesmo assim não se tem informações sobre sua biologia básica nessas áreas, sendo classificada como uma espécie com dados insuficientes (DD). Portanto, o presente trabalho teve como objetivo estimar a idade e crescimento; caracterizar aspectos da biologia reprodutiva e hábitos alimentares; e estudar a morfometria linear, geométrica e a estrutura populacional da raia A. narinari na costa da Paraíba e Pernambuco (07°30S; 34°49W / 07°47S; 34°51'W), Nordeste do Brasil. As amostras foram coletadas em maio e julho de 2015 e mensalmente de abril de 2016 a agosto de 2018 pela pesca artesanal da região. Os indivíduos foram identificados, sexados, fotografados, a largura do disco (LD, cm) e o peso total (PT, kg) foram tomados. Para o estudo do crescimento, a idade foi estimada através da contagem das bandas translúcidas das vértebras para 26 fêmeas (LD: 53,2 - 155 cm) e 24 machos (LD: 44,2 - 136 cm), assim sendo o modelo de crescimento com o melhor ajuste para ambos os sexos foi o de von Bertalanffy (fêmeas: L = 164,2; k = 0,18 e t0= -2,4; machos L = 153,3; k = 0,25 e t0= -1,9); e os indivíduos mais velhos foram uma fêmea com nove anos (LD: 155 cm) e um macho com cinco anos (LD: 136 cm). Para a biologia reprodutiva, com base na análise de estruturas macroscópicas sexuais secundárias, 27 fêmeas (LD: 53,2 - 155 cm) e 27 machos (LD: 39,5 - 141 cm) foram analisados, sendo observado que para as fêmeas o tamanho em que 50% da população atingiu a maturidade (L50) foi de 130,15 cm e para os machos foi de 116,42 cm. Os maiores valores médios do índice hepatossomático (IHS) e índice gonadossomático (IGS) para os machos ocorreram no terceiro bimestre do ano, e para as fêmeas ocorreu no último bimestre para o IHS e no quarto bimestre para o IGS. Em relação ao estudo da dieta, com base na análise dos conteúdos estomacais, a raia pintada apresentou uma dieta especialista baseada em presas do grupo Chordata e Mollusca, onde fêmeas se alimentaram principalmente de Teleostei (IRI %: 46,4) e Ascídia (IRI %: 27,9); e os machos de Ascídia (IRI %: 57,2) e Gastropoda (IRI %: 20,3). Para a morfometria linear, além da LD, mais 29 medidas morfométricas foram tomadas para comparação entre machos e fêmeas; as Análises de Covariância (ANCOVA) realizadas para os sexos separados demonstrou diferença significativa nas regressões lineares entre cada medida e a LD dos indivíduos. Para a morfometria geométrica foram utilizadas fotografias da vista dorsal e ventral dos condrocrânios de 38 indivíduos (15 machos e 23 fêmeas); e conforme a análise de Componentes Principais (PCA) e Discriminante houve diferença significativa entre os sexos em ambas as vistas dorsal e ventral. Finalmente, para estudar a estrutura populacional, 42 indivíduos foram utilizados e sequências concatenadas COI-Cytb-ND4 de genes mitocondriais foram analisados; a amostra apresentou 14 diferentes haplótipos, com diversidade hd= 0,646; baixos valores de índice de fixação (FST) por local; indicando fraca estrutura populacional. Na área de estudo a pesca artesanal de pequena escala captura espécimes de A. narinari de todas as idades e diferentes estágios de maturação durante todo o ano, portanto, o estudo pode contribuir para o manejo da pesca e conservação da espécie na região.

The spotted eagle ray, Aetobatus narinari, occurs in coastal and island tropical regions of the Atlantic Ocean. The recent taxonomic redefinition of the species has reduced the available literature on its biology and fishing, as well as made its conservation status (Near Threatened - NT) obsolete worldwide. In Brazil, this ray is caught by artisanal fisheries in several regions; even so there is no information about its basic biology in these areas, being classified as a Data Deficient species (DD). Therefore, the present work aimed to estimate age and growth; characterize aspects of reproductive biology and eating habits; and study the linear and geometric morphometry and population structure of the ray A. narinari in the coast of Paraíba and Pernambuco (07°30'S; 34°49'W / 07°47'S; 34°51'W), Northeast Brazil. The samples were collected in May and July of 2015 and monthly from April 2016 to August 2018 by artisanal fishing. The individuals were identified, sexed, photographed, the disc width (DW, cm) and total mass (TM, kg) were taken. For the study of growth, age was estimated by counting the translucent bands of the vertebrae for 26 females (DW: 53.2 - 155 cm) and 24 males (DW: 44.2 - 136 cm), so the growth model with the best fit for both sexes was von Bertalanffy (females: L = 164.2; k = 0.18 e t0= -2.4; males L = 153.3; k = 0.25 e t0= -1.9); and the older individuals were a nine-year-old female (DW: 155 cm) and a five-year-old male (DW: 136 cm). For reproductive biology, based on the analysis of secondary sex macroscopic structures, 27 females (DW: 53.2 - 155 cm) and 27 males (DW: 39.5 - 141 cm) were analyzed, females body size at which 50% of the individuals are mature (DW50) at 130.15 cm and males at 116.42 cm. The highest mean values of the hepatosomatic index (IHS) and gonadosomatic index (IGS) for males occurred in the third bimester of the year, and for females occurred in the last bimester for IHS and in the fourth bimester for IGS. Regarding the study of diet, based on the analysis of stomach contents, the Whitespotted Eagle Ray presented a specialist diet based on prey from the Chordata and Mollusca group, where females fed mainly on Fish (IRI %: 46.4) and Ascidians (IRI %: 27.9); and the males on Ascidians (IRI %: 57.2) and Gastropoda (IRI %: 20.3). For linear morphometry, besides DW, 29 additional morphometric measurements were taken for comparison between males and females; therefore the results of Covariance Analyses (ANCOVA) performed for the separate sexes showed a significant difference in linear regressions between each measurement and the DW of individuals. For the geometric morphometry, photographs of the dorsal and ventral view of the chondrochraniums of 38 individuals (15 males and 23 females) were used; and according to the statistical of Principal Component analysis and Linear Discriminant analysis there were significant differences for the sexes in both dorsal and ventral views. Finally, to study the population structure, 42 individuals were used and concatenated COI-Cytb-ND4 sequences of mitochondrial genes were analyzed; and the sample presented 14 different haplotypes, with diversity hd= 0.646; low fixation index (FST) values per site; indicating weak population structure. In the study area the small-scale artisanal fishery catches A. narinari specimens of all ages and different stages of maturity throughout the year, so the study can contribute to the management of fishing and conservation of the species in the region.