Secuencias del gen mitocondrial para identificación de especies de animales

Molecular markers based on mitochondrial DNA (mtDNA) have maternal inheritance and may be used to evaluate phylogenetic and taxonomic issues.The similarity diagnosis of living beings can be performed by genomic analysis of the 16S ribosomal RNA (16S rRNA). The aim of this study was to evaluate the molecular marker in 16S rRNA gene potential to identify different animal species and determine the degree of genetic similarity among individuals. DNA was extracted from 13 animals being 5 sheep, 4 cattle and 4 fish and the samples were amplified, purified and sequenced. The analysis were made with MEGA 5.10 and BLAST programs. The sequences identified on GenBank® showed that the animals belonged to the Ovis aries (sheep), Bos taurus (cattle), Prochilodus lineatus (curimba), Pseudoplatystoma corruscans (pintado), Pseudoplatystoma reticulatum (cachara) and Leporinus obtusidens (piau) species. The genetic distance between the different animal groups was 0,10 cattle/sheep, 0,66 cattle/fish and 0,65 sheep/fish. The construction of the phylogenetic tree has determined two different classes being Mammalia containing the subfamillies: Bovinae and Caprinae, and Actinopterygii containing the orders: Caraciforme (curimba and piau) and Siluriforme (pintado and cachara). The mtDNA molecular markers technology demonstrated the possibility of using it as a tool for species identification and differentiation thereby assisting the work of taxonomists. Thus, the partial sequencing of the 16S rRNA gene was sufficient for the identification of sheep, cattle and different species of fish based on the similarity of that gene in BLAST. Marcadores moleculares basados en el ADN mitocondrial (ADNmt) tienen herencia materna y se pueden utilizar para evaluar relaciones filogenéticas y taxonómicas. El diagnóstico de similaridad en los seres vivos se puede lograr mediante análisis genómico del ARN ribosomal 16S (16S rARN). El objetivo de este estudio fue evaluar el potencial de marcador molecular en el16S ARNr como una metodología para la identificación de las diferentes especies de animales y determinar el grado de similitud genética entre estos individuos. Para ello, el ADN se extrajo a partir de 13 animales, 5 ovinos, 4 bovinos y 4 peces, a continuación se amplificaron, se purificaron y se secuenciaron las muestras. Los análisis de las secuencias se realizaron en los programas MEGA 5.10 y BLAST. Las secuencias identificadas en GenBank® mostraron que los animales pertenecían a las espécies Ovis aries (ovino), Bos taurus (bovino), Prochilodus lineatus (curimba) Pseudoplatystoma corruscans (pintado), Pseudoplatystoma reticulatum (cachara) y Leporinus obtusidens (piau). La distancia genética entre los diferentes grupos de animales fue de 0,10 bovinos/ovinos, 0,66 bovinos/peces y 0,65 ovinos/peces. La construcción del árbol filogenético ha determinado dos clases distintas siendo Mammalia, que contiene las subfamilias: Bovinae y Caprinae, y Actinopterygii que contiene las órdenes: Caraciforme (curimba y piau) y Siluriforme (pintado y cachara). La tecnología de los marcadores moleculares del ADNmt demonstró la posibilidad de utilizarla como herramienta en la identificación y diferenciación de las especies asistiendo a la obra de los taxonomistas. Por lo tanto, la secuenciación parcial de la región del gen 16S rARN fue suficiente para la identificación de ovinos, bovinos y diferentes especies de peces sobre la base de la similitud del gen en programa BLAST

Sequências de gene mitocondrial para identificação de espécies animais

Molecular markers based on mitochondrial DNA (mtDNA) have maternal inheritance and may be used to evaluate phylogenetic and taxonomic issues. The similarity diagnosis of living beings can be performed by genomic analysis of the 16S ribosomal RNA (16S rRNA). The aim of this study was to evaluate the molecular marker in 16S rRNA gene potential to identify different animal species and determine the degree of genetic similarity among individuals. DNA was extracted from 13 animals being 5 sheep, 4 cattle and 4 fish and the samples were amplified, purified and sequenced. The analysis were made with MEGA 5.10 and BLAST programs. The sequences identified on GenBank® showed that the animals belonged to the Ovis aries (sheep), Bos taurus (cattle), Prochilodus lineatus (curimba), Pseudoplatystoma corruscans (pintado), Pseudoplatystoma reticulatum (cachara) and Leporinus obtusidens (piau) species. The genetic distance between the different animal groups was 0,10 cattle/sheep, 0,66 cattle/fish and 0,65 sheep/fish. The construction of the phylogenetic tree has determined two different classes being Mammalia containing the subfamillies: Bovinae and Caprinae, and Actinopterygii containing the orders: Caraciforme (curimba and piau) and Siluriforme (pintado and cachara). The mtDNA molecular markers technology demonstrated the possibility of using it as a tool for species identification and differentiation thereby assisting the work of taxonomists. Thus, the partial sequencing of the 16S rRNA gene was sufficient for the identification of sheep, cattle and different species of fish based on the similarity of that gene in BLAST.Marcadores moleculares baseados em DNA mitocondrial (mtDNA) possuem um padrão de herança materna podendo ser utilizados na avaliação de relações filogenéticas e taxonômicas. O diagnóstico de similaridade entre seres vivos pode ser realizado pela análise genômica do RNA ribossômico 16S (16S rRNA). O objetivo deste trabalho foi avaliar a potencialidade de marcador molecular na região 16S rRNA para identificação de diferentes espécies de animais e determinar o grau de similaridade genética entre estes indivíduos. Para isso, extraiu-se o DNA de 13 animais, sendo 5 ovinos, 4 bovinos e 4 peixes, em seguida as amostras foram amplificadas, purificadas e sequenciadas. As análises das sequências foram realizadas nos programas MEGA 5.10 e BLAST. As análises das sequências no GenBank® demonstraram que os animais pertenciam as espécies de Ovis aries (ovino), Bos taurus (bovino), Prochilodus lineatus (curimba), Pseudoplatystoma corruscans (pintado), Pseudoplatystoma reticulatum (cachara) e Leporinus obtusidens (piau). A distância genética entre os diferentes grupos animais foi de 0,10 bovinos/ovinos, 0,66 bovinos/peixes e de 0,65 ovinos/peixes. A construção da árvore filogenética permitiu determinar duas classes distintas sendo Mammalia, contendo as subfamílias: Bovinae e Caprinae, e Actinopterygii contendo as ordens: Caraciforme (curimba e piau) e Siluriforme (pintado e cachara). A tecnologia de marcadores moleculares de mtDNA demonstrou a possibilidade da sua utilização como ferramenta na identificação e diferenciação de espécies auxiliando o trabalho de taxonomistas. Assim, o sequenciamento parcial da região do gene 16S rRNA foi suficiente para a identificação de ovinos, bovinos e diferentes espécies de peixes com base na similaridade do gene no programa BLAST.Marcadores moleculares basados en el ADN mitocondrial (ADNmt) tienen herencia materna y se pueden utilizar para evaluar relaciones filogenéticas y taxonómicas. El diagnóstico de similaridad en los seres vivos se puede lograr mediante análisis genómico del ARN ribosomal 16S (16S rARN). El objetivo de este estudio fue evaluar el potencial de marcador molecular en el16S ARNr como una metodología para la identificación de las diferentes especies de animales y determinar el grado de similitud genética entre estos individuos. Para ello, el ADN se extrajo a partir de 13 animales, 5 ovinos, 4 bovinos y 4 peces, a continuación se amplificaron, se purificaron y se secuenciaron las muestras. Los análisis de las secuencias se realizaron en los programas MEGA 5.10 y BLAST. Las secuencias identificadas en GenBank® mostraron que los animales pertenecían a las espécies Ovis aries (ovino), Bos taurus (bovino), Prochilodus lineatus (curimba) Pseudoplatystoma corruscans (pintado), Pseudoplatystoma reticulatum (cachara) y Leporinus obtusidens (piau). La distancia genética entre los diferentes grupos de animales fue de 0,10 bovinos/ovinos, 0,66 bovinos/peces y 0,65 ovinos/peces. La construcción del árbol filogenético ha determinado dos clases distintas siendo Mammalia, que contiene las subfamilias: Bovinae y Caprinae, y Actinopterygii que contiene las órdenes: Caraciforme (curimba y piau) y Siluriforme (pintado y cachara). La tecnología de los marcadores moleculares del ADNmt demonstró la posibilidad de utilizarla como herramienta en la identificación y diferenciación de las especies asistiendo a la obra de los taxonomistas. Por lo tanto, la secuenciación parcial de la región del gen 16S rARN fue suficiente para la identificación de ovinos, bovinos y diferentes especies de peces sobre la base de la similitud del gen en programa BLAST

Identificação de contaminação microbiológica e potencial mutagênico de águas superficiais do município de Dourados, MS

The aim of the study was to evaluate the water quality of the Água Boa stream and the lakes of the parks Arnulpho Fioravante and Antenor Martins (Dourados - MS), through physico-chemical parameters, microbiological analysis and mutagenicity test to monitor interference from Antropic activities and water depuration capacity. Initially the water samples were processed for coliform analysis using the Most Probable Number (MPN) technique. For the research of Salmonella spp., it was performed the pre-enrichment and enrichment of the water sample and streaking in Xylose Lysine Deoxicolate (XLD) agar. For the evaluation of the mutagenicity, smear peripheral blood of the specimens Astyanax lacustris were collected from the places studied. The Arnulpho Fioravante Park had higher mean values for Electrical Conductivity (353.5 uS cm-2), Total Dissolved Solids (179.3 mg L-1) and the highest values for total coliforms. Salmonella spp. it was identified in the water in both parks. The highest numbers of micronucleus were observed in Água Boa stream. Considering microbiological contamination and higher micronucleus number, it’s relevant to highlight the effects of anthropic interferences in local biota. Therefore, it’s of great importance to intensify the supervision of the municipal legislation, in order to conserve and preserve these ecosystemsO objetivo do estudo foi avaliar a qualidade da água do córrego Água Boa e lagos dos parques Arnulpho Fioravante e Antenor Martins (Dourados – MS), por meio de parâmetros físico-químicos, análises microbiológicas e teste de mutagenicidade para monitorar a interferência proveniente de atividades antrópicas e a capacidade de depuração da água. Inicialmente as amostras de água foram processadas para análise de coliformes técnica do Número Mais Provável (NMP). Realizou-se o pré enriquecimento e enriquecimento das amostra e estriamento em Agar Desoxicolato Lisina Xilose (XLD) para pesquisa de Salmonella spp. Para avaliação da mutagenicidade foi realizado o esfregaço sanguíneo de espécimes de Astyanax lacustris coletados nos locais de estudo. O parque Arnulpho Fioravante apresentou as maiores médias para Condutividade Elétrica (353,5 uS cm-2), Sólidos Totais Dissolvidos (179,3 mg L-1) e os maiores valores para coliformes totais. Salmonella spp também foram identificadas nas águas provenientes dos Lagos dos parques. Os maiores números de micronúcleo foram observados no córrego Água Boa. Considerando a contaminação microbiológica e o número elevado de micronúcleo, torna-se relevante ressaltar os efeitos das interferências antrópicas na biota local. Sendo de grande importância a intensificação da fiscalização de legislações municipais vigentes para que haja a conservação e preservação desses ecossistemas

Genotoxic and mutagenic effects of polluted surface water in the midwestern region of Brazil using animal and plant bioassays

Abstract This study aimed to evaluate DNA damage in animal and plant cells exposed to water from the Água Boa stream (Dourados, Mato Grosso do Sul, Brazil) by using bioassays, and to identify the chemical compounds in the water to determine the water quality in the area. Through the cytotoxicity bioassay with Allium cepa, using micronucleus test, and comet assay, using Astyanax altiparanae fish, the results indicated that biological samples were genetically altered. Micronuclei were observed in erythrocytes of A. altiparanae after exposure to water from locations close to industrial waste discharge. The highest DNA damage observed with the comet assay in fish occurred with the exposure to water from locations where the presence of metals (Cu, Pb, Cd, Ni) was high, indicating the possibility of genotoxic effects of these compounds. Thus, these results reinforce the importance of conducting genotoxicity tests for developing management plans to improve water quality, and indicate the need for waste management before domestic and industrial effluents are released into the rivers and streams

Genotoxic and mutagenic effects of polluted surface water in the midwestern region of Brazil using animal and plant bioassays

Abstract This study aimed to evaluate DNA damage in animal and plant cells exposed to water from the Água Boa stream (Dourados, Mato Grosso do Sul, Brazil) by using bioassays, and to identify the chemical compounds in the water to determine the water quality in the area. Through the cytotoxicity bioassay with Allium cepa, using micronucleus test, and comet assay, using Astyanax altiparanae fish, the results indicated that biological samples were genetically altered. Micronuclei were observed in erythrocytes of A. altiparanae after exposure to water from locations close to industrial waste discharge. The highest DNA damage observed with the comet assay in fish occurred with the exposure to water from locations where the presence of metals (Cu, Pb, Cd, Ni) was high, indicating the possibility of genotoxic effects of these compounds. Thus, these results reinforce the importance of conducting genotoxicity tests for developing management plans to improve water quality, and indicate the need for waste management before domestic and industrial effluents are released into the rivers and streams