Diversidade bacteriana ruminal em bovinos Nelore
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Data
2014-07-01
Autores
Jesus, Raphael Barbetta de [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
O rúmen é um ecossistema aberto, no qual o alimento consumido pelos ruminantes é fermentado a ácidos graxos de cadeia curta e proteína microbiana, os quais servem respectivamente como fonte de energia e de proteína para o animal. Espécies de micro-organismos têm desenvolvido no rúmen uma série de interações complexas, o qual é um dos melhores exemplos de simbiose entre micro-organismos na natureza. Os métodos convencionais para a taxonomia baseada em técnicas de cultivo vêm sendo substituídas por técnicas moleculares, que são rápidas e mais precisas. O fundamento das técnicas moleculares está na sequência do 16S rRNA que fornece a classificação filogenética usadas na identificação e quantificação da comunidade bacteriana. Para avaliar a diversidade bacteriana ruminal neste estudo, foram utilizados 3 bovinos da raça Nelore, canulados no rúmen. As frações líquida e sólida do conteúdo ruminal foram processadas para extração de DNA metagenômico, após a extração foi verificado a quantidade e integridade das amostras. Em seguida, foi feita a PCR baseada nas regiões hipervariáveis V1 e V2 do 16S rRNA, e em sequência procedeu-se a construção da biblioteca e sequenciamento utilizando a plataforma Illumina. Os dados foram analisados pelos softwares MG-RAST e MOTHUR para filiações bacterianas. Aproximadamente 11.407.000 reads foram geradas com qualidade e 812 e 752 UTOs foram encontrados no nível de espécie e gênero respectivamente. Foram identificados 27 filos no conteúdo ruminal de bovinos Nelore através do sequenciamento do gene 16S rRNA pela plataforma Illumina. Os conhecimentos gerados a partir do presente estudo são informações primárias e primordiais para o entendimento da composição bacteriana ruminal. Assim, nos proporciona vislumbrar futuro promissor no desenvolvimento de novos métodos e tecnologias aplicáveis na nutrição animal
The rumen is an open ecosystem in which the food consumed by ruminant is fermented to yield short-chain fatty acids and microbial protein, that in turn serve as a source of energy and protein for the animal, respectively. Species of microorganisms into rumen have developed a series of complex interactions that become one of the best examples of symbiosis between microorganisms in nature. Conventional methods for microbial taxonomy based on culture techniques have been replaced by molecular techniques which are quickly and more accurate. The majority of phylogenetic molecular tools for bacteria classification are based on ribosomal 16S rRNA gene that provides resources for identification and also quantification of bacterial communities. In order to evaluate the ruminal bacterial diversity present in three Nelore cannulated cattle, the rumen content was fractionated in liquid and solid samples after its collect. Both parts were processed for metagenomic DNA extraction which in turn was evaluated according quantity and integrity parameters. Next stage consisted in PCR technique based on V1 and V2 hypervariable 16S rRNA regions to generate amplicons used for DNA sequencing performed at Illumina platform. Data were processed by MG-RAST and MOTHUR softwares to deduce bacterial affiliations. Approximately 11,407,000 reads were generated showing quality for indication of 812 and 752 OTUs at the level of species and genus, respectively. Twenty-seven phyla were successfully identified in Nellore ruminal contents by 16S rRNA sequencing in the powerful Illumina platform. The knowledge generated from this study are primary and essential for the wide understanding of rumen bacterial composition information. Thus, it provides us resources to be explored in a promising future focusing the development of new methods and technologies applied to animal nutrition
The rumen is an open ecosystem in which the food consumed by ruminant is fermented to yield short-chain fatty acids and microbial protein, that in turn serve as a source of energy and protein for the animal, respectively. Species of microorganisms into rumen have developed a series of complex interactions that become one of the best examples of symbiosis between microorganisms in nature. Conventional methods for microbial taxonomy based on culture techniques have been replaced by molecular techniques which are quickly and more accurate. The majority of phylogenetic molecular tools for bacteria classification are based on ribosomal 16S rRNA gene that provides resources for identification and also quantification of bacterial communities. In order to evaluate the ruminal bacterial diversity present in three Nelore cannulated cattle, the rumen content was fractionated in liquid and solid samples after its collect. Both parts were processed for metagenomic DNA extraction which in turn was evaluated according quantity and integrity parameters. Next stage consisted in PCR technique based on V1 and V2 hypervariable 16S rRNA regions to generate amplicons used for DNA sequencing performed at Illumina platform. Data were processed by MG-RAST and MOTHUR softwares to deduce bacterial affiliations. Approximately 11,407,000 reads were generated showing quality for indication of 812 and 752 OTUs at the level of species and genus, respectively. Twenty-seven phyla were successfully identified in Nellore ruminal contents by 16S rRNA sequencing in the powerful Illumina platform. The knowledge generated from this study are primary and essential for the wide understanding of rumen bacterial composition information. Thus, it provides us resources to be explored in a promising future focusing the development of new methods and technologies applied to animal nutrition
Descrição
Palavras-chave
Bovino, Nelore (Zebu), Bacterias, Rúmen, Rumen - Microbiologia
Como citar
JESUS, Raphael Barbetta de. Diversidade bacteriana ruminal em bovinos Nelore. 2014. iv, 33 p. Dissertação (mestrado) - Universidade Estadual Paulista Júlio de Mesquita Filho, Faculdade de Ciências Agrárias e Veterinárias, 2014.