Submitted by Irene Nascimento (irene.kessia@ufpe.br) on 2016-06-30T16:59:18Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Dissertação- Raphael Santana.pdf: 1198352 bytes, checksum: 2ddd13ab9df70bdaf76d80dd5d481f49 (MD5) / Made available in DSpace on 2016-06-30T16:59:19Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Dissertação- Raphael Santana.pdf: 1198352 bytes, checksum: 2ddd13ab9df70bdaf76d80dd5d481f49 (MD5) Previous issue date: 2015-02-24 / FACEPE / Actinobactérias são bactérias Gram-positivas formadoras de filamentos ramificados que se destacam pela produção de metabólitos secundários, como antimicrobianos, antitumorais, fitohormônios e corantes naturais. Diante disso, este trabalho teve o objetivo de determinar o potencial biotecnológico de actinobactérias da coleção UFPEDA contra Candida spp. Para avaliação da atividade biotecnológica, 32 actinobactérias foram testadas contra sete isolados clínicos de Candida spp. utilizando diferentes meios de cultura para o crescimento (ALA, ISP-4, AY, ISP-3) e temperaturas (37 ºC ou 45 ºC). No ensaio primário, foi evidenciada atividade apenas quando as actinobactérias foram cultivadas em meio ISP-3 a temperatura de 37 ºC. Dessas, apenas 2 linhagens (6,25 %) apresentaram atividade antifúngica com halo de inibição variando entre 11 mm e 18 mm. As duas linhagens produtoras de metabólitos secundários com atividades antifúngicas não apresentaram diferença estatística, por isso foi selecionada a linhagem G24 por apresentar um pigmento na cor vermelha para dar seguimento às análises. A fermentação da linhagem (G24) foi realizada para avaliar atividade antifúngica, biomassa e pH, durante 5 dias, utilizando diferentes meios (MPE, M1 e ISP-3),sendo tais parâmetros monitorados a cada 24 horas. O melhor meio para produção de metabólitos secundários foi ISP-3 fermentado durante 48 h em pH 7.0, sendo evidenciados halos de inibição de 13 a 18 mm e biomassa de 0,1 g/mL de meio. Estabelecidas às condições, foi realizada a extração do princípio bioativo, sendo evidenciada atividade apenas para biomassa quando extraído em acetato de etila. A concentração mínima inibitória (CMI) do extrato bruto variou de 125 μg/mL a 62,5 μg/mL para Candida spp testadas. Análise Cromatográfica do Extrato Bruto as frações semi-purificadas foram analisadas por cromatografia em camada delgada (CCD) sendo evidenciadas duas frações, uma com atividade antifúngica e outra um corante natural. As características morfológicas do isolado G24 foram analisadas por microscopia óptica, sendo observados esporos verticilados pertencente ao gênero Streptomyces. O gene 16S rDNA foi amplificado e enviado para sequenciamento, sendo identificada como Streptomyces sp. Diante destes resultados, podemos concluir que a linhagem (Streptomyces sp), isolado da rizosfera de Caesalpinia pyramidalis tul, do bioma Caatinga, apresenta uma significativa atividade antifúngica e necessita de estudos espectroscópios para caracterização do composto bioativo e do corante. / Actinobacteria are forming Gram-positive bacteria of branched filaments that stand out for the production of secondary metabolites such as antibiotics, antitumor, phytohormones and naturias dyes. Given this, this study aimed to determine the biotechnological potential of actinomycetes of UFPEDA collection against Candida spp. To evaluate the biotechnological activity, 32 actinomycetes were tested against seven clinical isolates of Candida spp., Using different culture media (ALA, ISP-4, AY, ISP-3) and temperatures (37 ° C and 45 ° C). In the primary test, activity was detected only when the actinomycetes were grown in ISP-medium 3 to a temperature of 37 ° C. Of these, only 2 strains (6.25%) showed antifungal activity with inhibition zone ranging between 11 mm and 18 mm. Fermentation of strain (G24) was used to evaluate antifungal activity, biomass and pH for 5 days, using different media (MPE M1 and ISP-3), these parameters being monitored every 24 hours. The best medium for production of secondary metabolites ISP-3 was fermented for 48 h at pH 7.0, being evidenced inhibition zones 13 to 18 mm and biomass of 0.1 g / ml medium. Established conditions, the extraction of bioactive principle has been performed and demonstrated activity only when biomass extracted into ethyl acetate. The minimum inhibitory concentration (MIC) of the crude extract ranged from 125 mg / mL to 62.5 mg / mL for Candida spp tested. In chemical prospecting, semi-purified fractions were analyzed by thin layer chromatography (TLC) was evidenced two fractions, one with antifungal activity and one with a natural dye. The morphological characteristics of isolated G24 were analyzed by optical microscopy, observed verticilados spores belonging to the genus Streptomyces. The 16S rDNA gene was amplified and sent to sequencing, being identified as Streptomyces sp. Given these results, we can conclude that the line (G24) (Streptomyces sp), isolated from the rhizosphere of Caesalpinia pyramidalis tul, the Caatinga biome, presents a significant antifungal activity and requires spectroscopes studies to characterize the bioactive compound and the dye.

Isopods in the genus Tylos are distributed in tropical and subtropical sandy intertidal beaches throughout the world. These isopods have biological characteristics that are expected to severely restrict their long-distance dispersal potential: (1) they are direct developers (i.e., as all peracarids, they lack a planktonic stage); (2) they cannot survive in the sea for long periods of immersion (i.e., only a few hours); (3) they actively avoid entering the water; and (4) they are restricted to the sandy intertidal portion that is wet, but not covered by water. Because of these traits, high levels of genetic differentiation are anticipated among allopatric populations of Tylos. We studied the phylogeographic patterns of Tylos in the northern East Pacific region between southern California and central Mexico, including the Gulf of California. We discovered high levels of cryptic biodiversity for this isopod, consistent with expectations from its biology. We interpreted the phylogeographic patterns of Tylos in relation to past geological events in the region, and compared them with those of Ligia, a co-distributed non-vagile coastal isopod. Furthermore, we assessed the usefulness of the shape of the ventral plates of the fifth pleonite for distinguishing genetically divergent lineages of Tylos in the study area. Finally, mitochondrial phylogenenetic analyses to identify the most appropriate outgroup taxa for Tylos in the study area, which included 17 of the 21 currently recognized species, provided important insights on the evolutionary history of this genus.