The graduation theses theme was to determinate changes of initial activity, total activity and Rubisco enzyme content in spinach leaves. Ascertained activities and Rubisco enzyme content were determined from extracts of spinach leaves, which were collected during two distinctively climatic different days. Both activities were determined by spectrometric method and content of enzyme Rubisco by SDS-PAGE method. The findings were compared with findings determined in earlier done works.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-12-13T12:21:07Z No. of bitstreams: 1 isabelmartinelecorrea.pdf: 473111 bytes, checksum: fef90be37cc2eb87e8dcbc88e8d3a4a5 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-12-15T14:08:57Z (GMT) No. of bitstreams: 1 isabelmartinelecorrea.pdf: 473111 bytes, checksum: fef90be37cc2eb87e8dcbc88e8d3a4a5 (MD5) / Made available in DSpace on 2016-12-15T14:08:57Z (GMT). No. of bitstreams: 1 isabelmartinelecorrea.pdf: 473111 bytes, checksum: fef90be37cc2eb87e8dcbc88e8d3a4a5 (MD5) Previous issue date: 2007-02-23 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Os protozoários ciliados do rúmen integram a microbiota ruminal e junto das bactérias, fungos e protozoários flagelados participam da digestão fermentativa nos ruminantes. Entretanto, o papel desempenhado por estes microorganismos no metabolismo do hospedeiro apresenta pontos controversos os quais se devem em parte, à complexidade da dinâmica populacional e às relações estabelecidas com o hospedeiro e demais microorganismos ruminais. Objetivou-se caracterizar as populações de protozoários ciliados no rúmen de ovinos mestiços Santa Inês, verificando os efeitos da estacionalidade e aspectos relacionados às variações diurnas destas populações. Foram utilizados cinco ovinos mestiços Santa Inês, fistulados no rúmen, mantidos em pastagem natural de caatinga, no estado de Pernambuco, Brasil. As coletas foram realizadas em julho de 2005, período correspondente à estação chuvosa, e em janeiro de 2006, durante a estação seca. A composição botânica da dieta dos animais foi determinada através da análise da extrusa. Os horários de coleta das amostras de conteúdo ruminal seguiram o tempo de soltura dos animais, sendo a primeira coleta antes de se soltar os animais (zero hora) e às duas, quatro, seis, oito e dez horas após o animal estar em campo, sendo as amostras fixadas em formalina 18,5% (v/v). A dieta alimentar dos ovinos durante a estação chuvosa foi predominantemente composta por espécies vegetais herbáceas e na estação seca por espécies arbustivas o que retrata o efeito da xvi estacionalidade no hábito alimentar dos ovinos. Foram identificados e quantificados organismos dos gêneros Dasytricha, Diplodinium, Diploplastron, Entodinium, Eodinium, Elytroplastron, Enoploplastron, Epidinium, Eremoplastron, Eudiplodinium, Isotricha, Metadinium e Ophryoscolex, sendo os gêneros Enoploplastron e Eodinium observados apenas na estação chuvosa. Diplodinium, Elytroplastron, Metadinium e Ophryoscolex não apresentaram variações na concentração de suas populações em função da estação do ano. Observou-se a predominância de organismos do gênero Entodinium nas estações chuvosa e seca; entretanto, na estação seca houve significativo decréscimo na concentração deste gênero, e aumento de outros entodiniomorfos e de isotriquídeos. O número total de protozoários foi significativamente reduzido durante a estação seca. Diploplastron e Entodinium apresentaram variações na concentração de suas populações em função do horário de amostragem. O número de ciliados em divisão não apresentou correlação com o horário de amostragem, sendo que a estabilidade das taxas de divisão parece contribuir para a manutenção das concentrações dos ciliados no rúmen. O pH ruminal apresentou correlação negativa com o tempo de amostragem, não estando correlacionado com o número de protozoários, provavelmente porque as variações observadas compreenderam valores considerados ideais para o estabelecimento e manutenção das populações de protozoários no rúmen. Foi constatado o comportamento predatório de Elytroplastron sobre Enoploplastron, Epidinium e Entodinium; além de canibalismo entre organismos do gênero Elytroplastron. Evidências do comportamento alimentar foram obtidas através do registro da ingestão de fibras e formação de grânulos de amido no citoplasma, o que evidencia a atividade celulolítica e amilolítica atribuída a estes microorganismos no ambiente ruminal. / The ciliate protozoan of the rumen are part of the ruminal microbiota, and together with bacteria, fungus and flagellate protozoa, they participate of the fermentative digestion of the ruminants. However, there are controversial aspects about the hole of these microorganisms it the metabolism of the host, in part due to the complexity of the population dynamics, and also to the interrelations between protozoan, the host, and other ruminal microorganisms. The objective of this work was to characterize the ciliate protozoa populations in the rumen of Santa Inez crossbred sheep, verifying the effects of the seasonality and also aspects related to the diurnal changes of these populations. It was used five sheep fistulated in the rumen, maintained in natural pasturage of caatinga (scrubland) vegetation at the state of Pernambuco, Brazil. The samples were collected in july 2005 (rainy season) and january 2006 (dry season). The composition of animals diet was determined through the extrusa analysis. The samples were obtained at intervals of two hours, being the first sample took before the releasing of the animals, in a total of six samples. The samples were fixed in formalin at 18, 5% (v/v). The feed composition of the sheep during the rainy season was predominantly composed of herbaceous species, and at the dry season by species shrub, denoting the effect of the seasonality on the sheep feed habits. It was identified and quantified Dasytricha, Diplodinium, Diploplastron, Entodinium, Eodinium, xviii Elytroplastron, Enoploplastron, Epidinium, Eremoplastron, Eudiplodinium, Isotricha, Metadinium e Ophryoscolex. Eodinium and Enoploplastron genders were observed just at the rainy season. Diplodinium, Elytroplastron, Metadinium and Ophryoscolex didn’t show variations in the population concentration as a function of the season. It was observed the predominance of the gender Entodinium at both seasons; however, at the dry season it was noted significant decrease in the concentration of this gender, and increase in greater ophryoscolecidae and Isotrichidae. The total number of protozoa was significantly reduced during the dry season. Diploplastron and Entodinium showed variations in their populations concentrations, due to the time of sampling. The number ciliates undergoing division was not correlated with the sampling times. Stable division rate appears to contribute to maintenance of ciliates concentrations in the rumen. The ruminal pH was negatively correlated with sampling times, and was not correlated with the number of protozoa, probably because the variations observed included parameters considered ideal for establishment and maintenance protozoa populations of rumen. It was observed predatory behavior of Elytroplastron on Enoploplastron, Epidinium and Entodinium; besides cannibalism between organisms of the Elytroplastron gender. The registration of fiber ingestion and starch granules formation at the cytoplasm indicates cellulolytic and amilollytic activities, attributed to these microorganisms in the ruminal environment.

In this diploma thesis, the diurnal changes of initial and total Rubisco activity and Rubisco enzyme content in beech (Fagus sylvatica) were studied under conditions of ambient (A) CO2 concentration (350 µmol.mol-1) and elevated (E) CO2 concentration (700 µmol.mol-1) during the day. Samples were taken on July 8th (from 10:00 to 21:30), on July 9th (from 04:00 to 12:00) and then on July 22th (from 04:00 to 21:30). The initial and total activity were measured spectrophotometrically and the activation level was calculated from the ratio of initial and total activities. Rubisco enzyme content was determined by SDS-PAGE method and the initial and total specific activity were calculated from the ratio of initial or total activity and Rubisco enzyme content. In our experiment no statistically significant difference was found between Rubisco activities in beeches cultivated under conditions of ambient CO2 concentration and elevated CO2 concentration, so any down-regulation of Rubisco activity did not appear under the influence of ambient CO2 concentration. Diurnal changes of Rubisco activities showed only statistically nonsignificant fluctuation. After daylight no significant increase of total Rubisco activity was observed, which demonstrates the absence or the immaterial effect of CA1P night inhibitor in beech. Intensive fluctuation of the activation level (40–90 %) proves the regulation of photosynthesis during the day via Rubisco enzyme carbamylation. Rubisco enzyme content in beech cultivated under conditions of ambient CO2 concentration was lower than that one in beech cultivated under conditions of elevated CO2 concentration. Then down-regulation of Rubisco enzyme content is presented under conditions of ambient CO2 concentration. Before daylight Rubisco enzyme content showed an indistinctive minimum on the level of statistical significance, during the day only nonrelevant fluctuation was noticed.