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Previous issue date: 2010-02-23 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior, CAPES, Brasil. / The increasing demand for biological processes alternative, environmentally friendly and
efficient in converting lignocellulosic material, expanding their application potential for
agribusiness, motivates researches worldwide. Thus, organisms isolated in nature, in specific
ecosystems, become increasingly important because of their physiological and metabolic
diversity, which gives them a great potential in the development of biotechnological processes
of interest to society. The aim of this study was to assess the microbial community associated
with the intestinal tract of millipede Trigoniulus corallinus and bioprospecting for
microorganisms with cellulolytic capacity. The millipedes were collected and incubated with
litter in diets of grass (Paspalum notatum) and ?sabia? (Mimosa caesalpinifolia). Sampling
occurred at 15, 30, 45, and 75 days of incubation. The intestinal tract of five individuals was
removed, sectioned the posterior third, processed and stored in ultrasound. DNA from
microbes associated with the intestinal tract, litter and coprolite was extracted, and DGGE
analysis using 16S rDNA, DGGE group actinomycetes, and it was evaluated the presence of
nifH genes. The 16s gene analysis by DGGE revealed a microbial diversity conditioned by the
diet offered to 45 days. After this period, this effect was no longer visible. The community
associated with coprolites and the type of litter was distributed in separate clusters of samples
from the intestinal tract. This effect was not observed in the community assessment of
actinomycetes, where the big difference for division of groups was the diet. The animals fed
on grass litter showed a diverse community, and they were not influenced by time or
compartmentalization. The samples associated with litter and coprolites were 80% similar to
samples from the intestinal tract. In millipedes fed with material form Mimosa caesalpinifolia,
the result was different, the samples of litter and coprolites where 50% similar to the intestinal
tract. All samples had nifH genes detected by polymerase chain reaction. Samples collected at
45 days were also inoculated in mineral minimum medium of Busnell-Hass added carboxymethyl-cellulose (CMC) as sole carbon source. Colonies were evaluated for their ability to
breakdown cellulose enzyme and 15 had an index greater than 1. The isolate that showed the
highest rate (3.65) was subjected to further analysis. The microscope observation suggested
that this was not an isolated but a complex of microorganisms acting on the degradation of
cellulose. There is evidence of BNF in the intestinal tract of the millipede and microorganisms
proliferated in CMC through the proper amplification of nifH genes and proliferation in
medium within nitrogen. The community of prokaryotes was influenced by the diet offered to
the community up to 45 days, and the actinomycetes community was conditioned by the diet.
It was possible to isolate microorganisms and complexes of microorganisms with cellulolytic
capacity, with great potential in the search for environmentally friendly technologies in
generating agrobioenergy. / A crescente demanda por processos biol?gicos alternativos, ambientalmente favor?veis e
eficientes na transforma??o de material ligninocelul?sico, ampliando seu potencial de
aplica??o agroindustrial, estimula pesquisas em todo o mundo. Assim, microrganismos
isolados na natureza, em ecossistemas espec?ficos, tornam-se cada vez mais importantes pela
sua diversidade metab?lica e fisiol?gica, que lhes confere grande potencialidade no
desenvolvimento de processos biotecnol?gicos de interesse ? sociedade. O objetivo deste
trabalho foi avaliar a comunidade microbiana associada ao trato intestinal do dipl?pode
Trigoniulus corallinus e a bioprospec??o de microrganismos com capacidade celulol?tica. Os
dipl?podes foram coletados e incubados em dietas com serrapilheira de grama batatais
(Paspalum notatum) e sabi? (Mimosa caesalpinifolia). As amostragens aconteceram aos 15,
30, 45 e 75 dias de incuba??o. O trato intestinal de cinco indiv?duos foi removido e
seccionado o ter?o posterior tratado em ultrasom e estocado. Procedeu-se a extra??o de DNA
da microbiota associada ao trato intestinal, serrapilheira e copr?lito, com an?lise por DGGE
utilizando o gene 16S rDNA, DGGE para grupo actinomicetos e avalia??o da presen?a de
genes nifH. A an?lise do gene 16s por DGGE revelou diversidade microbiana condicionada
pela dieta oferecida at? os 45 dias. Ap?s este per?odo o efeito n?o foi mais vis?vel. A
comunidade associada aos copr?litos e ao tipo de serrapilheira distribui-se em grupamentos
separados das amostras oriundas do trato intestinal. O mesmo n?o foi observado na avalia??o
da comunidade de actinomicetos, onde o grande diferencial para divis?o de grupos foi a dieta.
Os animais alimentados com serrapilheira de grama mostraram uma comunidade diversa e
n?o influenciada pelo tempo ou compartimentaliza??o. As amostras associadas ? serrapilheira
e aos copr?litos foram 80% similares ?s do trato intestinal. Nos dipl?podes alimentados com
sabi?, o resultado foi diferente, sendo as amostras de serrapilheira e copr?litos 50% similares
?s do trato intestinal. Todas as amostragens tiveram genes nifH detectados via PCR. Amostras
coletadas aos 45 dias foram tamb?m inoculadas em meio mineral m?nimo de Busnell-Hass
adicionado de carboxi-metil-celulose (CMC) como ?nica fonte de carbono. Os microrganismos isolados foram avaliados quanto ? capacidade de degrada??o de celulose e 15
apresentaram ?ndice enzim?tico maior que 1. O isolado com o maior ?ndice (3,65) foi alvo de
outras an?lises. A visualiza??o em microsc?pio sugeriu que n?o se tratava de um isolado e
sim de um complexo de microrganismos atuando na degrada??o da celulose. H? evidencias de
FBN no trato intestinal do dipl?pode e microrganismos proliferados em meio CMC pela boa
amplifica??o de genes nifH e prolifera??o em meio com aus?ncia de nitrog?nio. A
comunidade de procariotos foi influenciada pela dieta oferecida at? os 45 dias e a comunidade
de actinomicetos foi condicionada em fun??o da dieta. Foram isolados microrganismos e
complexos de microrganismos com capacidade celulol?tica, com grande potencial para a
busca de tecnologias ambientalmente sustent?veis na gera??o de agrobioenergia.
| Identifer | oai:union.ndltd.org:IBICT/oai:localhost:jspui/1735 |
| Date | 23 February 2010 |
| Creators | Passos, Samuel Ribeiro |
| Contributors | Xavier, Gustavo Ribeiro, Correia, Maria Elizabeth Fernandes, Anjos, Lucia Helena Cunha dos, Soares, Lu?s Henrique de Barros |
| Publisher | Universidade Federal Rural do Rio de Janeiro, Programa de P?s-Gradua??o em Agronomia e Ci?ncia do Solo, UFRRJ, Brasil, Instituto de Agronomia |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFRRJ, instname:Universidade Federal Rural do Rio de Janeiro, instacron:UFRRJ |
| Rights | info:eu-repo/semantics/openAccess |
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