Naturalized Breeds in Brazil: Reports on the Origin and Genetic Diversity of the Pantaneiro Sheep

Brazil has several breeds of sheep, including animals that developed from breeds brought by settlers soon after their discovery. Over the years, these animals were under the process of natural selection, resulting in breeds that are considered naturalized. The Pantaneiro sheep shows rusticity and ability of adaptation to tropical climate regions and tolerance or resistance to disease and parasites. Molecular tools have marked the discovery of the origin and domestication processes of a wide variety of species, using both nuclear and mitochondrial molecular markers. These tools have aided in the understanding of evolutionary relationships, taxonomies, and demographics of various species and provided support to identify the most important areas for conservation programs, in addition to assisting in the analysis of genetic diversity in domestic, wildlife and endangered species. Researches using these tools show the importance of exploiting the potential of the genetic diversity found in locally adapted livestock. So far, a few studies were performed to observe that Pantaneiro sheep served as maternal basis for the origin of other breeds reared. Moreover, it is possible to suggest an European origin for the sheep populations studied; therefore, more studies using more markers are needed, so that it is possible to prove their origin

Discriminação alélica em ovinos naturalizados do Pantanal Sul-Matogrossense por meio de marcadores microssatélites

The molecular biology techniques that are used in allelic discrimination for individual and sheep breedscharacterization are important tools in breeding programs and conservation of genetic resources. The use ofmicrosatellite markers allows allelic differentiation, which in turn allows us to infer the genetic variability of samplepopulations. The study aimed to test the sensitivity and efficiency of fluorescent capillary electrophoresis, using microsatellite primers, for allelic discrimination of the Crioulo breed from Pantanal sul-matogrossense, as well asverify the possibility of using the products of sequencing in genetic variability analysis. For this test, were used bloodsamples from Pantaneira breed sheep. The allelic discrimination of eight microsatellites was determined by capillaryelectrophoresis in automatic sequencer and the results analyses were performed on the programs CERVUS andDendro-UPGMA. The results indicated the possibility of using this technique for the individual genotyping of all locitested in electrophoretic analysis and its potential to allelic discrimination even in case of difference between twopairs of bases between the alleles. The resulting dendrogram based on the distance matrix by the UPGMA assemblymethod, indicated medium similarity coefficient of 0.72 in the group of animals. It was concluded that there is theviability and efficiency of the microsatellite molecular markers technique using capillary electrophoresis for allelicdiscrimination and the utility of results for studies of genetic variability, paternity diagnosis and characterization ofthe Crioulo sheep herd from Pantanal sul-matogrossense. As técnicas de biologia molecular que são utilizadas na discriminação alélica para caracterização individual e de raçasde ovinos são ferramentas importantes nos programas de melhoramento genético e de conservação de recursosgenéticos. A utilização de marcadores moleculares microssatélites possibilita a diferenciação alélica, que por sua vezpermite inferir a variabilidade genética de populações amostrais. O estudo teve como objetivo testar a sensibilidade ea eficiência da eletroforese capilar fluorescente, utilizando oligonucleotídeos iniciadores de microssátelites, paradiscriminação alélica da raça crioula do pantanal sul-matogrossense, bem como verificar a possibilidade de utilizaçãodos produtos do sequenciamento em análises de variabilidade genética. Para o referido teste foram utilizadas amostrasde sangue provenientes de ovinos da raça pantaneira. A discriminação alélica dos oito microssatélites foi determinadapor meio de eletroforese capilar em sequenciador automático e as análises dos resultados foram realizadas nosprogramas CERVUS e Dendro-UPGMA. Os resultados indicaram a possibilidade da utilização da técnica para agenotipagem individual de todos os loci testados numa corrida eletroforética e a sua potencialidade paradiscriminação alélica mesmo em situações de diferença de dois pares de bases entre os alelos. O dendrogramaresultante baseado na matriz de distância pelo método de agrupamento UPGMA, indicou coeficiente de similaridademédio de 0.72 no grupo de animais. Foi possível concluir que existe viabilidade e eficiência da técnica de marcadoresmoleculares microssatélites utilizando eletroforese capilar para discriminação alélica e a utilidade dos resultados paraestudos de variabilidade genética, diagnóstico de paternidade e caracterização do rebanho de ovinos crioulos doPantanal Sul-mato-grossense

Avaliação de protocolos para extração de DNA Genômico de sangue Bovino

Basic studies on DNA extraction techniques are very important for the success of scientific papers in the field ofmolecular biology. The extraction and purification of nucleic acids are critical steps for establishing further geneticanalysis. The objective of this study was to evaluate the efficacy of different protocols for DNA extraction by deter- mining the quantity and quality of extracted genetic material and the possibility of amplification by PCR. We didDNA extraction and PCR of ten bovine blood samples. The test results obtained by spectrophotometry indicated thatthe quantity and quality of genomic DNA were considered satisfactory in all protocols for PCR. However, there wasa statistically significant difference between the parameters measured, both in quantity and in quality (p <0.01). Theextraction protocol using whole blood was more efficient in terms of time and quality; there was no degradation inall processes of extraction. It was also demonstrated that the possibility of amplification of the region of exon 2 ofthe leptin gene in extracted DNA exists.Estudos básicos sobre técnicas de extração de DNA são de extrema importância para o sucesso de trabalhos científicos no campo da biologia molecular. A extração e a purificação de ácidos nucleicos constituem etapas fundamentaispara o estabelecimento de posteriores análises genéticas. Objetivou-se neste trabalho avaliar a eficácia de diferentesprotocolos de extração de DNA por meio da determinação de quantidade, qualidade e a possibilidade de amplificação por meio de PCR do material genético extraído. Utilizou-se amostras de sangue de dez animais, que foramsubmetidas à extração de DNA e, logo após à reação em cadeia pela polimerase (PCR). Os resultados das análisesobtidas por nanofotometria indicaram que a quantidade e a qualidade do DNA genômico foram consideradas satisfatórias, em todos os protocolos, para a realização da PCR, mas houve diferença estatística significativa entre osparâmetros analisados, tanto na quantidade quanto na qualidade (p < 0.05) do DNA obtido. O protocolo de extraçãoutilizando sangue total foi mais eficiente quanto ao tempo e a qualidade do DNA extraído, entretanto em todos osprocessos de extração houve ausência de degradação. Também foi demonstrado a possibilidade de amplificação daregião do exon 2 do gene da leptina de bovinos no DNA extraído

Allelic discrimination in naturalized ovine from Pantanal Sul-Matogrossense by means of microsatellite markers

The molecular biology techniques that are used in allelic discrimination for individual and sheep breeds characterization are important tools in breeding programs and conservation of genetic resources. The use of microsatellite markers allows allelic differentiation, which in turn allows us to infer the genetic variability of sample populations. The study aimed to test the sensitivity and efficiency of fluorescent capillary electrophoresis, using microsatellite primers, for allelic discrimination of the Crioulo breed from Pantanal sul-matogrossense, as well as verify the possibility of using the products of sequencing in genetic variability analysis. For this test, were used blood samples from Pantaneira breed sheep. The allelic discrimination of eight microsatellites was determined by capillary electrophoresis in automatic sequencer and the results analyses were performed on the programs CERVUS and Dendro-UPGMA. The results indicated the possibility of using this technique for the individual genotyping of all loci tested in electrophoretic analysis and its potential to allelic discrimination even in case of difference between two pairs of bases between the alleles. The resulting dendrogram based on the distance matrix by the UPGMA assembly method, indicated medium similarity coefficient of 0.72 in the group of animals. It was concluded that there is the viability and efficiency of the microsatellite molecular markers technique using capillary electrophoresis for allelic discrimination and the utility of results for studies of genetic variability, paternity diagnosis and characterization of the Crioulo sheep herd from Pantanal sul-matogrossense

Discriminação aléica em ovinos naturalizados do Pantanal Sul-Matogrossense por meio de marcadores microssatélies

The molecular biology techniques that are used in allelic discrimination for individual and sheep breeds characterization are important tools in breeding programs and conservation of genetic resources. The use of microsatellite markers allows allelic differentiation, which in turn allows us to infer the genetic variability of sample populations. The study aimed to test the sensitivity and efficiency of fluorescent capillary electrophoresis, using microsatellite primers, for allelic discrimination of the Crioulo breed from Pantanal sul-matogrossense, as well as verify the possibility of using the products of sequencing in genetic variability analysis. For this test, were used blood samples from Pantaneira breed sheep. The allelic discrimination of eight microsatellites was determined by capillary electrophoresis in automatic sequencer and the results analyses were performed on the programs CERVUS and Dendro-UPGMA. The results indicated the possibility of using this technique for the individual genotyping of all loci tested in electrophoretic analysis and its potential to allelic discrimination even in case of difference between two pairs of bases between the alleles. The resulting dendrogram based on the distance matrix by the UPGMA assembly method, indicated medium similarity coefficient of 0.72 in the group of animals. It was concluded that there is the viability and efficiency of the microsatellite molecular markers technique using capillary electrophoresis for allelic discrimination and the utility of results for studies of genetic variability, paternity diagnosis and characterization of the Crioulo sheep herd from Pantanal sul-matogrossense.As técnicas de biologia molecular que são utilizadas na discriminação alélica para caracterização individual e de raças de ovinos são ferramentas importantes nos programas de melhoramento genético e de conservação de recursos genéticos. A utilização de marcadores moleculares microssatélites possibilita a diferenciação alélica, que por sua vez permite inferir a variabilidade genética de populações amostrais. O estudo teve como objetivo testar a sensibilidade e a eficiência da eletroforese capilar fluorescente, utilizando oligonucleotídeos iniciadores de microssátelites, para discriminação alélica da raça crioula do pantanal sul-matogrossense, bem como verificar a possibilidade de utilização dos produtos do sequenciamento em análises de variabilidade genética. Para o referido teste foram utilizadas amostras de sangue provenientes de ovinos da raça pantaneira. A discriminação alélica dos oito microssatélites foi determinada por meio de eletroforese capilar em sequenciador automático e as análises dos resultados foram realizadas nos programas CERVUS e Dendro-UPGMA. Os resultados indicaram a possibilidade da utilização da técnica para a genotipagem individual de todos os loci testados numa corrida eletroforética e a sua potencialidade para discriminação alélica mesmo em situações de diferença de dois pares de bases entre os alelos. O dendrograma resultante baseado na matriz de distância pelo método de agrupamento UPGMA, indicou coeficiente de similaridade médio de 0.72 no grupo de animais. Foi possível concluir que existe viabilidade e eficiência da técnica de marcadores moleculares microssatélites utilizando eletroforese capilar para discriminação alélica e a utilidade dos resultados para estudos de variabilidade genética, diagnóstico de paternidade e caracterização do rebanho de ovinos crioulos do Pantanal Sul-mato-grossense

Evaluation protocols for the extraction of genomic DNA from Bovine blood

Basic studies on DNA extraction techniques are very important for the success of scientific papers in the field of molecular biology. The extraction and purification of nucleic acids are critical steps for establishing further genetic analysis. The objective of this study was to evaluate the efficacy of different protocols for DNA extraction by determining the quantity and quality of extracted genetic material and the possibility of amplification by PCR. We did DNA extraction and PCR of ten bovine blood samples. The test results obtained by spectrophotometry indicated that the quantity and quality of genomic DNA were considered satisfactory in all protocols for PCR. However, there was a statistically significant difference between the parameters measured, both in quantity and in quality (p <0.01). The extraction protocol using whole blood was more efficient in terms of time and quality; there was no degradation in all processes of extraction. It was also demonstrated that the possibility of amplification of the region of exon 2 of the leptin gene in extracted DNA exists