Resumo A Coleção de Culturas de Microalgas de Água-Doce (CCMA-UFSCar), localizada na Universidade Federal de São Carlos tem um importante papel na sustentação da pesquisa brasileira em microalgas, provendo material biológico, substratos e treinamento de pessoal para uma grande fração dos projetos atuais nessa área. Entretanto, os recentes esforços para expandir o catálogo de culturas atingiram um limite máximo em praticidade logística para a manutenção de culturas metabolicamente ativas. Buscando reduzir os custos do regime de manutenção das culturas, foi-se iniciado o estabelecimento de um banco criopreservado para a manutenção em longo prazo desses organismos, minimizando os gastos com manuseio do material, visto que culturas congeladas com sucesso podem, em teoria, serem mantidas por prazos indeterminados, ainda com a vantagem da estabilidade das características nas culturas. Testes iniciais foram realizados com 93 linhagens, incluindo organismos de diferentes grupos taxonômicos mantidos na coleção de culturas. As maiores taxas de sucesso foram observadas nos organismos menores, como as pequenas algas verdes, enquanto os organismos maiores e mais complexos obtiveram sucesso limitado. Em geral, para as linhagens testadas, acima de 70% dos organismos apresentaram resposta positiva ao processo.

Abstract The Culture Collection of Freshwater Microalgae (CCMA-UFSCar, Coleção de Culturas de Microalgas de Água-Doce), based at the Universidade Federal de São Carlos, plays an important role in underpinning Brazilian microalgal research, providing biological materials, substrates and training personnel for a large proportion of the past and current projects in this area. However, recent efforts to expand the culture catalogue/holdings have reached a ceiling because of the logistical practicality of maintaining actively growing cultures. In order to reduce the costs associated with the maintenance regime of cultures, efforts were initiated on the establishment of a cryopreserved biobank for long term maintenance of cultures, thus minimizing the efforts associated with handling of material, as successfully frozen cultures, in theory, could be maintained effectively indefinitely, with the advantage of the stability of cells characteristics. Initial tests were performed on 93 strains including exemplar taxa across the different taxonomic groups in the collection catalogue. The highest levels of success were achieved for the smaller taxa, such as the small green algae; while for the larger and more complex organisms limited to no success was obtained. For the strains tested, over 70% had positive good levels of post-thaw viability and regenerated phenotypically normal cultures.

O objetivo deste trabalho foi avaliar as respostas das células de Haematococcus pluvialis ao processo de indução à carotenogênese, sob estresse luminoso e nutricional. As células foram aclimatadas durante 15 dias em meio WC, com aeração com ar atmosférico sintético filtrado e fluxo de 100 mL min-1, intensidade luminosa de 50 µmol fótons m-2 s-1, fotoperíodo de 12 horas e temperatura de 23ºC. Foram comparados dois tratamentos: cultivo nas condições descritas, mas com aumento da intensidade luminosa para 350 µmol fótons m-2 s-1 ; e cultivo nas mesmas condições do tratamento anterior, mas com aeração contendo 4% de CO2. Os tratamentos foram conduzidos em triplicata, durante dez dias. Com a adição de CO2 e o incremento da iluminação, observou-se aumento da razão carotenoides/clorofila e da biomassa celular. As células cessaram a divisão no segundo dia de estresse, quando o nitrato se tornou limitante, e aumentaram significativamente seu biovolume. A excreção de carbono orgânico e a concentração de astaxantina aumentam em resposta à adição de CO2. O estresse por intensidade luminosa, aliado à adição de CO2, otimiza a carotenogênese em H. pluvialis e aumenta a produção de astaxantina.

The objective of this work was to evaluate the responses of Haematococcus pluvialis cells to the carotenogenesis induction process, under light and nutrition stress. Cells were acclimated during 15 days in WC medium, with aeration with synthetic, filtered atmospheric air and flow rate of 100 mL min-1, light intensity of 50 µmol photons m-2 s-1, photoperiod of 12 hours, and temperature of 23ºC. The following two treatments were compared: cultivation under the described conditions, but with increase of light intensity up to 350 µmol photons m-2 s-1 ; and cultivation under the same conditions as the previous treatment, but with aeration containing 4% CO2. The treatments were done in triplicate, during ten days. With the addition of CO2 and the increment in lighting, an increase was observed in the carotenoids/chlorophyll ratio and cell biomass. Cells stopped dividing on the second day of stress, when nitrate became limiting, and significantly increased their biovolume. The excretion of organic carbon and the concentration of astaxanthin increase in response to the addition of CO2. Stress by light intensity combined with CO2 addition optimizes carotenogenesis in H. pluvialis and increases astaxanthin production.

In this work, the fatty acid quantity and composition of six freshwater microalgae and soybean grains was determined by direct transesterification and gas chromatography analysis. The results showed that all the freshwater microalgae species presented a higher quantity of fatty acid than soybean grain. Choricystis sp. (A) provides 115% more fatty acids per gram of biomass than soybean grain. With regard to the fatty acid composition, Choricystis sp. (A) showed an adequate proportion of saturated and unsaturated fatty acids, with lower quantity of polyunsaturated fatty acids and, akin to some marine microalgae, constitutes an alternative raw material for biodiesel production.

Glycolytic activities of eight enzymes in size-fractionated water samples from a eutrophic tropical reservoir are presented in this study, including enzymes assayed for the first time in a freshwater environment. Among these enzymes, rhamnosidase, arabinosidase and fucosidase presented high activity in the free-living fraction, while glucosidase, mannosidase and galactosidase exhibited high activity in the attached fraction. The low activity registered for rhamnosidase, arabinosidase and fucosidase in the attached fraction seemed contribute to the integrity of the aggregate and based on this fact, a protective role for these structures was proposed. The presented enzyme profiles and the differences in the relative activities probably reflected the organic matter composition as well as the metabolic requirements of the bacterial community, suggesting that bacteria attached to particulate matter had phenotypic traits distinct from those of free-living bacteria.

O estudo dos carboidratos dissolvidos e sua relação com as algas abundantes no reservatório de Barra Bonita, foi realizado em coletas mensais (junho de 2002 a janeiro de 2004), em seis profundidades na coluna d’água, cujos valores variaram dependendo da estação do ano e do nível do reservatório, num ponto fixo, localizado 3 km a montante da barragem. Aproximadamente 90% dos carboidratos dissolvidos neste reservatório foram constituídos por carboidratos poliméricos hidrolisáveis (polissacarídeos ou oligossacarídeos). A análise de variância detectou variabilidade entre as concentrações dos carboidratos dissolvidos apenas em escala temporal (p < 0,05), sendo que no eixo vertical, as concentrações destes açúcares apresentaram-se homogêneas (p > 0,05). A composição monossacarídica dos açúcares encontrados no reservatório foi galactose, ramnose, glicose e fucose. Na maioria dos meses amostrados, as concentrações de carboidratos dissolvidos, a maioria deles poliméricos, ocorreram concomitantemente ao registro de florescimentos das espécies abundantes, principalmente cianofíceas, que provavelmente foram as principais fontes de polissacarídeos extracelulares para o sistema.

Dissolved carbohydrates and abundant algae of Barra Bonita Reservoir were studied monthly from June 2002 to January 2004 in six depths whose values vary depending on the seasons and the water level at a sampling site located 3 km upstream the Reservoir dam. Nearly 90% of the dissolved carbohydrates were constituted of polymeric hydrolysable carbohydrates (polysaccharides or oligosaccharides). Analysis of variance detected variability among concentrations of dissolved carbohydrates only in temporal scale (p < 0.05), whereas in vertical axis the concentrations of these sugars were uniform (p > 0.05). The monosaccharide composition of the sugars found in the reservoir was galactose, rhamnose, glucose e fucose. During the sampling period, higher concentrations of dissolved carbohydrates, mostly polymeric, occurred together with large phytoplanktonic blooms, predominantly cyanobacteria, which probably were the main source of extracellular polysaccharides in the system.

O objetivo deste trabalho foi investigar o efeito caotrópico do íon Li+ na cápsula polissacarídica da microalga colonial Ankistrodesmus gracilis. Esta microalga possui uma cápsula extensa e contínua que envolve completamente as células. Neste estudo, foram utilizados a técnica de Ressonância Paramagnética Eletrônica (RPE) e o marcador de spin - Tempo (2,2,6,6-tetrametilpiperidine-1-oxil), de natureza não reativa. A técnica de RPE é utilizada para monitorar o tempo de difusão dos marcadores de spin através da cápsula polissacarídica, parede e membrana celular, até o interior da célula, onde eles perdem suas propriedades paramagnéticas. O íon Li+ utilizado tem a propriedade de provocar a desorganização estrutural da cápsula e, em conseqüência, a modificação do tempo de difusão do marcador de spin. Observou-se que os tempos de decaimento do sinal paramagnético nas células tratadas com Li+ são sempre menores do que nas células não expostas ao Li+ e que os tempos de difusão diminuem com o aumento da concentração de LiCl . As fotomicrografias das algas mostram que a partir da concentração de LiCl 1,0 M, as cápsulas são totalmente retiradas, resultando em células que se aglutinam formando grumos. Também foi constatada a morte das células acima quando expostas a concentração de LiCl acima de 1,0 M.

The aim of this study was to investigate the role of chaotropic effect of Li+ on the polysaccharide sheath of the microalga Ankistrodesmus gracilis (Chlorophyceae, Chlorococales) in the selective permeability and transport of molecules in the interior of the cell. The colonial microalga Ankistrodesmus gracilis has a relatively large and continuous mucilaginous sheath. The Electron Paramagnetic Resonance (EPR) was applied using the spin label- Tempo 2,2,6,6-tetramethylpiperidine-1-oxyl of non-reactive nature to study its diffusion coefficients across the polysaccharide envelope. The EPR technique is used to monitor the spin labels diffusion time across the polysaccharide capsule, cell wall and membrane into the interior of the cell, where they lose their paramagnetic properties. The lithium ion (Li+) has the property to structurally disorganize the polysaccharide capsule and the spin label diffusion changes producing different spin label time decay. The time decays of cells subjected to Li+ treatment are faster than the cells without this ion usage and the decay time decreases with increasing Li+ concentration. The photomicrographies show the complete removal of the cell capsule from LiCl 1,0 M upward and as a result, the algal cells agglutinate producing cell clumps, and the algal cells are clearly died.

Foram efetuadas várias medidas de excreção de matéria orgânica dissolvida (MOD), utilizando-se populações naturais de fitoplâncton, provenientes da Enseada do Flamengo - litoral Norte do Estado de São Paulo - e enriquecidas artificialmente com nutrientes. A quantidade absoluta da MOD excretada, de um modo geral, acompanhou a quantidade de carbono total assimilado. Entretanto, nas populações com deficiência de nitrogênio, ocorreu um aumento das taxas relativas de excreção, não só devido à diminuição da assimilação total de carbono, mas, efetivamente, tambem ao aumento da quantidade absoluta de MOD excretada.

Measurements of excreted dissolved organic matter (DOM) were carried out twice (in simmer and winter) using natural phytoplankton populations from "Enseada do Flamengo" (Lot. 23° 30'S-Long. 045°069'W) and enriched artificially with nutrients. The bioassays were carried out inoculating samples of an integral sea water in ten liters flasks, incubated "in situ". Measures were taken in populations growing in: integral " f " media, "f" - N, "f" - P, "f" - (H + P), "f" - (Vit. + Metal traces), and in a control without enrichment. The absolute amount of excreted DOM generally followed the quantity of total carbon assimilated. In the meantime, with the populations with nitrogen deficiency occurred an increase of relatives rates of excretion, not only due to the total carbon assimilation diminution, but effectively also to the absolute increase of the quantity of excreted DOM.