Dissertações em Biotecnologia (Mestrado) - PPGBIOTEC/ICB

URI Permanente para esta coleçãohttps://repositorio.ufpa.br/handle/2011/6090

O Mestrado em Biotecnologia teve início em 2011 e funciona no Programa de Pós-Graduação em Biotecnologia (PPGBIOTEC) do Instituto de Ciências Biológicas (ICB) da Universidade Federal do Pará (UFPA).

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Acesso aberto (Open Access)
Acúmulo de colesterol por Mycobacterium smegmatis como possível modulador da biossíntese de lipomanana e lipoarabinomanana
(Universidade Federal do Pará, 2015-02-26) SANTOS, Ana Cristina Doria dos; SENA, Chubert Bernardo Castro de; http://lattes.cnpq.br/8620752020290438
Mycobacterial cell wall is a hallmark of Mycobacterium genus, constituted of bioactive lipids and glycoconjugates: phosphatidylinositol mannoside (PIMs), lipomannan (LM) and lipoarabinomannan (LAM). The chronic infection inside lung macrophages is related with cholesterol accumulation into host cells as an alternative source of carbon and energy to maintain the bacilli with its physiological functions. To understand the activity of these immunomodulatory glycoconjugates during adaptation under the unusual environment inside the host infected cells, the present work propose to investigate the possible modulation of LM/LAM biosynthesis in Mycobacterium smegmatis (saprophytic) after culture in minimal medium (MM), supplemented with glycerol and/or cholesterol. Our results showed that saprophytic bacilli are able to accumulate the cholesterol and change the bacterial physiology due to slow growth and restrict cell density. Furthermore, the cholesterol consumption decreased the accumulation of PIMs and promoted morphological changes and bacterial aggregates, even maintaining the cell wall with its specific physic-chemistry characteristic (alcohol-acid resistance). The most impressive change after cholesterol consumption was the LAM biosynthesis, which showed distinct electrophoresis migration, compatible to high molecular weight of LAM from non-saprophytic bacilli (from 25 – 30 KDa to 30 – 50 KDa). These results showed that cholesterol consumption, when utilized as principal alternative of carbon and energy source, is able to induce physiological changes in mycobacteria, mainly in LAM biosynthesis, one of the most immunoregulatory molecules of cell wall. Ours data suggest that similar changes in mycobacteria may occur inside the granuloma, and that changes may help the evolution of tuberculosis to chronic and multibacillary form, hallmarked by immunodeficient aspect against the bacilli.
Acesso aberto (Open Access)
Análise da expressão gênica de pacientes com adenocarcinoma gástrico associado ao vírus Epstein-Barr
(Universidade Federal do Pará, 2024-08) SILVA, Valéria Cristiane Santos da; MOREIRA, Fabiano Cordeiro; http://lattes.cnpq.br/5745396559731337
Infection with the Epstein-Barr virus (EBV) is one of the risk factors for gastric cancer (GC). Epstein-Barr is a virus with oncogenic activity and was the first to be associated with malignant diseases such as lymphomas and carcinomas. Thus, EBV detection can be performed both by in situ hybridization and, currently, by using RNA sequencing techniques to identify the presence of viral genes in samples. In this context, aiming to better understand the EBV-positive subtype, this study proposed a molecular classification based on RNA sequencing of 76 tumor samples from patients diagnosed with GC. RNA sequencing of the samples was performed, and molecular classification was done using the Kraken2 software. Of the 76 samples, 8 were considered positive according to the adopted method. Subsequently, to understand the different mechanisms by which EBV might be acting in gastric cancer, we analyzed the patterns of human gene expression in samples classified as positive and negative. In our study, there are approximately 834 differentially expressed genes, of which 92 have an AUC greater than 0.85. These genes are associated with tumor progression, cellular metabolism, and innate and adaptive immunity. Additionally, viral genes expressed in the positive samples were evaluated, and we found manifestations of both lytic phase and latent phase genes. Finally, our study presents an efficient strategy for the molecular classification of EBV-positive
Acesso aberto (Open Access)
Análise e caracterização do genoma mitocondrial: um estudo de caso de Cyprinodon sp. e Arapaima gigas
(Universidade Federal do Pará, 2025-05) MACEDO, Daralyns Borges; RAMOS, Rommel Thiago Juca; http://lattes.cnpq.br/1274395392752454; https://orcid.org/0000-0002-8032-1474
The advancement of next-generation sequencing technologies has enabled large-scale characterization of mitochondrial genomes across various species. However, even among fish — the group with the highest number of sequenced mitochondrial genomes — only 9.7% of valid species are represented in genetic databases. This gap limits conservation programs, which rely on genetic data to guide management strategies. The genus Cyprinodon, composed of several threatened species restricted to isolated habitats, is particularly vulnerable to habitat loss and interspecific hybridization. In this context, the present study aimed to sequence and characterize the mitochondrial genome of Cyprinodon sp., in order to clarify phylogenetic relationships and evolutionary aspects of the Cyprinodontidae family. The sample was sequenced using the Illumina HiSeq platform, assembled with Megahit software, and analyzed using tools such as MitoZ, Circos, ClustalW, MrBayes, and Figtree. The complete mitogenome comprises 16,500 bp and contains 37 genes. Concurrently, Arapaima gigas, an iconic Amazonian species historically important to fisheries, is currently threatened with extinction and classified as “Data Deficient” by the IUCN. In the Tocantins River, heavily impacted by human activities, fishers from 25 communities reported the near disappearance of the species, with only one recent sighting on Jaracuera Island. The disappearance of A. gigas has severe economic consequences for local communities. Without collaboration between fishery managers and local populations, the species faces an imminent risk of regional extinction, further exacerbated by the lack of data on its fishing dynamics in northern Brazil.
Acesso aberto (Open Access)
Análise morfológica e molecular dos filamentos das nadadeiras pélvicas do peixe pulmonado Lepidosiren paradoxa
(Universidade Federal do Pará, 2015-12-18) LIMA, Sérgio Queirós; SCHNEIDER, Igor; http://lattes.cnpq.br/4453558334183860
The Lepidosiren paradoxa species belongs to Dipnoi order, along with two genre and are considered true lungfishes. Adult males of L. paradoxa differ from females by the presence of filaments in the pelvic fin. These filaments resemble those found in gills of fish and neotenic salamanders. These filaments are grow and become vascularized during the reproduction period. In this work we tested the hypothesis that the pelvic filament of L. paradoxa share morphological and molecular characteristics with gill filaments. Thus, we performed morphological and molecular analyses of the filaments of the pelvic fins between the dry and rainy seasons. Morphological analysis was perfomed through hematoxylin and eosin (HE) and scanning electron microscopy (SEM). Finally, we quantified gene expression of gill markers by RT-PCR using pectoral fin as reference. The mean length and standard deviation of the filaments during rainy and dry season were 4,31mm of ± 0.186 and 1,63mm ± 0.104, respectively. In the SEM images, we observed with a few cell microvilli and/or microplicae and some smaller cells. In the HE analyzes we found that the filaments have uniform epithelium formed with four layers of cells being filled by connective tissue and finally become more vascularized the rainy season. Molecular analysis through RT-PCR did not show any change when compared to the pectoral fin and between the stations. In conclusion, despite morphological and molecular similarities between gill filaments and pelvic fin filaments, our findings do not support a role for L. paradoxa pelvic filaments in gas exchange.
Acesso aberto (Open Access)
Análise proteômica de cultura macrófagica tratada com ácido kójico
(Universidade Federal do Pará, 2023-07) ALMEIDA, Heyder Coutinho; SANTOS, Agenor Valadares; http://lattes.cnpq.br/9530734927662735; https://orcid.org/0000-0002-2690-2841
Kojic acid (AK) is a natural fungal secondary metabolite, produced by some species of Aspergillus, Penicillium and Acetobacter, through the direct biotransformation of carbon substrates such as glucose, sucralose, ethanol, among others. AK was originally isolated in Japan, in 1907, from Aspergillus oryzae mycelia, it is used in the cosmetic industry in the form of gels and soap and is used to obtain the effect of whitening the skin by inhibiting the enzyme tyrosinase, reducing the production of formation of melanin, it also blocks the formation of pigmentation and evens out the skin tone. It is also applied as a food additive for the prevention of enzymatic browning, as it is a metal chelator and a potent antioxidant. Although AK has numerous biological functions, only recently studies on in vitro proliferation and cytotoxicity have been reported, but still limitedly regarding its effect on immune cells. Macrophages are among the most important defense cells that specifically recognize and respond to foreign bodies, apoptosis cells and pathogens. Through the activation process, there is an increased proliferation of macrophages, which undergo various morphological changes, such as an increase in dissemination and adhesion abilities, phagocytosis activity, ROS generation, antigen presentation and cytokine production. According to the classification of biological functions, our results revealed that in both Kojic acid concentrations, the 14 expressed proteins have a predicted function related to cell cycle and redox processes.
Acesso aberto (Open Access)
AutoAssemblyD software para submissão e gerenciamento de montagem de genomas a partir de modelos XML
(Universidade Federal do Pará, 2014-01-24) VERAS, Adonney Allan de Oliveira; SILVA, Artur Luiz da Costa da; http://lattes.cnpq.br/7642043789034070
Technologies for second-generation sequencing provided a major breakthrough of the genome, making its use a landmark that has revolutionized biology. These platforms are characterized by a reduction in sequencing time, high data production and low cost per base sequenced, however, these devices produce data mostly consist of short readings which represents a major challenge for reconstruction of the genome due to this new feature readings of computational tools had to be developed to accomplish the task of assembling their example we Velvet, AllPaths, Abyss, SOAPdenovo2, Edena. However, most of these applications are executed through command lines extended and composed of several parameters must follow the standard syntax to use, because in case of errors in the syntax is the possibility of not obtaining the best result, with the aim of solve this problem we present the AutoAssemblyD that besides providing the use of these assemblers through a graphical interface also enables the management of these executions remotely.
Acesso aberto (Open Access)
Avaliação do viés GC em plataformas de sequenciamento de nova geração
(Universidade Federal do Pará, 2015-03-05) PINHEIRO, Kenny da Costa; RAMOS, Rommel Thiago Jucá; http://lattes.cnpq.br/1274395392752454
The emergence of high throughput sequencing (HTS) platforms increased the amount of data making feasible to obtaining complete genomes. Despite the advantages and the throughput produced by these platforms, the high or low genomic coverage in the regions of the genome can be related to GC content. This GC bias may affect genomic analyzes and the genomic/transcriptomic analysis based on de novo and reference approach. In addition, the ways to evaluate the GC bias should be fit to data with different profiles of the GC vs coverage relationship, such as linear and quadratic. Thus, this work proposes the use of Pearson's Correlation Coefficient (r) to analyze the correlation between GC content and coverage, allowing to identify the strength of linear correlation and detect nonlinear associations, beyond identify a relationship between GC bias and sequencing platforms. The positive and negative signs of r also allow us to infer directly and inversely proportional relationships, respectively. To evaluate the bias, we used the data of Corynebacterium pseudotuberculosis obtained from different sequencing technologies to identify if the CG bias is related to used platforms.
Acesso aberto (Open Access)
Bioprospecção de leveduras com potencial para a produção de carotenoides presentes em uma microrregião do bioma Amazônia
(Universidade Federal do Pará, 2024-11) LUCAS, David Cristian Rodrigues; CHISTÉ, Renan Campos; http://lattes.cnpq.br/0583058299891937; https://orcid.org/0000-0002-4549-3297
Carotenoids are bioactive compounds used as natural pigments and are found in plants, animals, algae and microorganisms. Some of these compounds are precursors of vitamin A and have benefits to human health such as strengthening the immune system and reducing the risk of chronic degenerative diseases. In industry, the production of carotenoids is obtained mainly by chemical synthesis or through algae and plant extracts. However, with the growing concern about utilization of chemical additives in food by many consumers, the interest in obtaining them through biotechnological processes and diverse microorganisms may synthesize them, such as yeasts. In this work, an exploration study was carried out focused on molecular identification of carotenoid-producing yeasts available in a microregion of Amazonia biome, with the aim of highlighting promising microorganisms for future industrial applications. Samples were collected from leaves, flowers, soil, and bark trees, among them, 4 promising strains were isolated and identified from phyllosphere of philodendron (Philodendron hederaceum): Rhodosporidiobolus ruineniae (PH-18(I)), Rhynchogastrema noutii or Hannaella pagnoccae (dPH-8(I)), Leucosporidium egoroviorum or Lyomyces sp. (dPH-8(II)), and Rhodotorula diobovata or Rhodotorula glutinis (PH-22(II)). The strain PH-18(I) showed a biomass yield of 0.26% to 0.76% and total carotenoid content of 139.70 μg/g of dry biomass. The strain dPH-8(I) showed a biomass yield of 0.43% to 0.96 % and total carotenoid content of 54.26 μg/g. The strain dPH-8(II) demonstrated a biomass yield of 0.35% to 1.12% and total carotenoid content of 52.71 μg/g, while the strain PH-20(II) demonstrated a biomass yield of 11.63 % to 23.67 % and total carotenoid content of 44.98 μg/g. The carotenoids identified in this study were 13Z-β-carotene, (all-E)-β-carotene and 9Z-β-carotene, being (all-E)-β-carotene the majority. Results found in this study highlight a promising perspective for future applications of carotenoid-producing yeasts in the Amazonia biome, being an excellent alternative to replace the process of obtaining carotenoids by synthetic means or by exploitation of vegetal sources.
Acesso aberto (Open Access)
Busca e classificação sistemática das proteínas oxigenases com ferro não hêmico em plantas
(Universidade Federal do Pará, 2015-03-06) SOUSA, Kellen Rayanne Matos de; DARNET, Sylvain Henri; http://lattes.cnpq.br/4586614214029929
Nonheme iron oxygenase proteins shares a conserved domain consisting of eight histidines, and they can be found in eukaryotes and prokaryotes organisms and participate in important pathways of lipid biosynthesis. To understand the evolutionary relationship among these proteins, we performed comparative and phylogenetic analyzes in prokaryotes and eukaryotes that allowed a classification of this family, nonexistent until now. The search of sequences resulted in a collection of 448 proteins, belonging to 58 organisms previously selected. The multiple alignment made with MAFFT (BLOSUM 62; L-INS-i) showed the presence of three histidine-rich motifs, with conserved spacing among them. The classification made with CLANS software has generated 28 clusters through of similarity among sequences. Two clusters contain sequences that had no similarity to proteins already characterized, and 48 sequences were not assigned to any of the 28 clusters. In the collection, 119 sequences are derived from plants, distributed in 7 clusters corresponding to C4 methysterol monooxygenase, C5 sterol desaturase, fatty acid hydroxylase, sphingolipid C4 monooxygenase, aldehyde decarbonylase, β-carotene hydroxylase and Acyl-ACP desaturase functions. Phylogenetic analysis using maximum likelihood method with PhyML tool showed the formation of well-defined groups that were similar to generated by CLANS. These results start to fill a gap existing so far about the evolutionary relationship and classification of nonheme iron oxygenases. Also, suggest that within this family there are still proteins with unknown functions, reinforcing the need for more studies of functional characterization.
Acesso aberto (Open Access)
Caracterização antigênica, físico-química e biológica do vírus BE AR 701405, obtido a partir de mosquitos da espécie Psorophora (Jan) ferox, capturados em Altamira – Pará
(Universidade Federal do Pará, 2014-03-26) ARAÚJO, João Batista dos Santos; LEAL, Élcio de Souza; http://lattes.cnpq.br/1158983666415285
The BE AR 701405 virus was isolated from a pool of Psorophora (Jan.) ferox mosquitoes captured in the municipality of Altamira, Para state, Northern Brazil, in 2006. The objective of this study was the physicochemical, biological and antigenic characterization of the BE AR 701405 virus in order to provide data for it taxonomic classification. Newborn mice inoculated by intracerebral route showed evidences of neurological manifestations, such as chill and motor disorders after the infection with the virus BE AR 701405. Cytophatic effect (CPE) was in infected Aedes albopictus C6/36 cells was not clearly observed, however in VERO cells CPE was observed after 48 hors post infection. The virus load was determined in 10-4,1 LD50/ 0.02 mL and the titer after DCE analysis was calculated in 10−2,6 LD50/0,02 mL. The BE AR 701405 virus reacted antigenically with Pixuna virus hiperimmune serum. In conclusion, the studied vírus was sensible to the DCA solvent suggesting that it is an enveloped virus. Furthermore, by serology this virus was identified as a member of the group A, genus Alphavirus, family Togaviridae, more closely related to Pixuna virus. In addition, newborn mice, as well as C6/36 and VERO cells demonstrated to be sensible to the infection by the BE AR 701405 vírus. Further studies are needed to better understand the antigenic relationship between the BE AR 701405 virus and Pixuna virus.
Acesso aberto (Open Access)
Cianotoxinas em ostras e em águas de cultivo da costa norte do Brasil
(Universidade Federal do Pará, 2016-08-31) CABRAL, Jardecicléia Patrícia da Silva; SCHNEIDER, Maria Paula Cruz; http://lattes.cnpq.br/3901112943859155
Cyanobacteria are photosynthetic prokaryotic organisms and under conditions favorable to their growth can form blooms. They are also capable of producing cyanotoxins, secondary metabolites that have a toxic effect on eukaryotic organisms including man. As part of phytoplankton the cyanobacteria participate in the food chains, thus the ingestion of shellfish, that have been continuously fed toxic cyanobacteria and accumulated the toxins in their tissues generates the possibility of transfer to the man through their consumption bringing serious risks to their health. There is still very little research on the contamination (by cyanotoxins) of shellfish intended for human consumption. Therefore, this study was to verify the occurrence of cyanotoxins in culture waters and in the tissue of oysters destined for human consumption. The water and oyster samples were collected from two municipalities located in the northeast of Pará. The cyanotoxins research was performed using High Performance Liquid Chromatography (HPLC). The cyanobacteria present in the water, isolated and cultured from oyster shells were identified. No microcystins and saxitoxins were detected in water samples and oyster extracts from both growing sites. However, three genera of cyanobacteria - Aphanizomenon, Oscillatoria and Phormidium were identified in Curuçá and three genera of cyanobacteria in São Caetano de Odivelas - Aphanothece, Oscillatoria and Phormidium, all known to contain toxic species. The presence of saxitoxins in extracts from cyanobacterial cultures was detected from the oyster shells of both growing sites, indicating the presence of toxic species despite the absence of bloom. In this study, it was verified that the cultivation of oyster sites are suitable for consumption, free of contamination by microcystins and saxitoxins, however, it is necessary to implement specific legislation and the monitoring of these crops in order to ensure consumer health.
Acesso aberto (Open Access)
Co-cultivo de embriões bovinos com células-tronco adultas derivadas de tecido adiposo
(Universidade Federal do Pará, 2014-09-05) NASCIMENTO, Hamilton Silva do; MIRANDA, Moysés dos Santos; http://lattes.cnpq.br/3354029928888919
The in vitro embryo production (IVP) is a biotechnology used to increase the reproductive potential of genetically superior animals, embryos produced in vitro are inferior in quality when compared in vivo. Techniques trying improve the rates of in vitro produced embryos. One technique is the co-culture system using somatic cells that remove toxic metabolites and protect against oxidative stress. Mesenchymal stem cells derived from adipose tissue (CTA) are multipotent cells that secrete growth factors and cytokines. The stem cells were used in in vitro co-culture of bovine embryos at different concentrations in order to improve the protocol PIVE. CTAs underwent differentiation into three mesenchymal lineages, and was performed immunophenotyping of specific membrane markers of MSCs. The cleavage rate was assessed on the second day after fertilization and blastocyst rate on the seventh day when they were stored for counting the total number of cells and gene expression. The results were analyzed by ANOVA, T-test and post-test of Fisher, adopting a significance level of 5%. Treatment of co-cultivation with CTAs significantly influenced the formation of the blastocyst, the total number of cells in embryos and correlated gene expression pluripotency and carbohydrate metabolism. These results showed increased production rate and quality of embryos produced in vitro co-culture with CTAs compared to co-culture with granulosa cells. These results also indicate that the constant presence of CTAs in co-culture is superior to conditioning with CTAs. Checked the effects of CTAs can occur through soluble factors or via exosomes secreted by CTAs. Future studies are needed to clarify the possible cause for positive effects observed in this work by the CTAs in co-cultivation.
Acesso aberto (Open Access)
Detecção de potenciais corticais antecipatórios em sinais de eletroencefalografia (EEG) durante a condução de carros
(Universidade Federal do Pará, 2015-03-16) SANTOS, Fredson Carmo dos; CARVALHO, Schubert Ribeiro de; http://lattes.cnpq.br/1496976331707751; GOMES, Bruno Duarte; http://lattes.cnpq.br/4932238030330851
The recognition of the driver’s intention from electroencephalographic signals (EEG) may be useful in the development of brain computer interface (BCI) to be used in synergy with intelligent vehicles. This can be beneficial to improve the quality of interaction between the driver and the car, for example, providing a response from the smart car aligned with the intention of the driver. In this study, the anticipation is considered as the cognitive state that leads to specific actions while driving a car. Therefore, we propose to investigate the presence of anticipatory patterns in EEG signals while driving vehicles to determine two specific actions (1) left and (2) turn right, a few milliseconds before such actions take place. An experimental protocol was proposed to record EEG signals of 5 individuals as they operate a virtual reality simulator non-invasive - it was designed for this experiment - which simulates driving a virtual car. The experimental protocol is a variant of the paradigm of contingent negative variation (CNV) with Go and Nogo conditions in virtual reality training system. The results of this study indicate the presence of anticipatory patterns observed in slow cortical potentials in the time domain (medium EEG signal) and the frequency (Power Spectra and phase coherence). This opens a range of possibilities in the development of BCI systems - based on anticipatory signals - that connect the driver to the intelligent vehicle favoring a decision-making to assess the intentions of drivers may eventually prevent accidents while driving.
Acesso aberto (Open Access)
Efeito do meio condicionado por células tronco mesenquimais derivadas de tecido adiposo na maturação de oócitos bovinos e posterior desenvolvimento embrionário in vitro
(Universidade Federal do Pará, 2016-07-29) VALENTE, Juliana Vieitas; MIRANDA, Moysés dos Santos; http://lattes.cnpq.br/3354029928888919
Stem cells are known for their property of self-renewal, differentiation into various cell lineages and immunomodulatory capabilities, in addition to express a large number of adhesion molecules, extracellular matrix proteins, cytokines and growth factor recptors, allowing interactions with other cells. They can be isolated from various tissues, but adult stem cells derived from adipose tissue stroma (MSC-d) have the advantage of being easely isolated, high yield and low morbidity. Based on this, the study hypothesis is to verify the effect of MSC-ad conditioned medium use in in vitro maturation on development rates of bovine embryos. To this end, bovine MSC ad were isolated then cultivated until the passage three (P3) in DMEM culture medium, supplemented with sodium bicarbonate, 10% fetal bovine serum (FBS), 50mL / mL gentamicin. When cultures reached 70% confluency, the medium was replaced with TCM199 supplemented with 0.2 mM pyruvate, 50 ul / ml gentamicin and 10% FBS (IVM medium). The MSC-ad were conditioned with medium TCM199 supplemented for a period of 0 (Control Group), 24 (COND-24 group), 48 (COND-48h group) and 72 hours (COND group-72h), wherein the end of each period, the supernatant medium was removed, filtered, aliquoted and frozen at -20 0 C. The medium was thawed only on MIV day, and supplemented with 0.5 uL of PMSG (concentration 7 IU / ml) and 0.5 uL of HCG. Slaughterhouse ovaries were punctured to obtain oophurus cumulus complexes (COCs) were matured for 24 hours in microdrops of conditioned mediums under mineral oil them incubated in an incubator at 5% CO 2 at 38.5 ° C. Nuclear maturation assessment was performed with 22 hours, then held 15 parthenogenetic activation of only oocytes that exhibited polar body concentration of at least 50 uM ionomycin for 5 minutes, inactivation with TCM199-Hepes 0.03g saturated with BSA, followed by incubation for 3 hours microdroplets 2mM 6-DMAP. After this period, the parthenogenesis were transferred as microdrops to a culture medium in vitro. development rates were evaluated in the second and seventh days after activation. The results were analyzed by ANOVA and Fisher's post-test adopting a significance level of 5%. For groups: Experimental CONTROL, COND-24, COND-48h and 72h-COND, we obtained the following results respectively: 83.7, 77.7, 81.4 and 76.1% of nuclear maturation; 87.5, 86.9, 74 and 80.3 and 23.8% cleavage, 27.5, 18 and 19.6% of blastocyst formation. There was no statistical difference between condinioned experimental groups (p<0.05). However, blastocyst rates were lower when compared with fresh IVM medium (42.1%) (p<0.05). This suggests that the efficiency was affected by the conditioning of the medium or freezing. Future studies should be conducted to assess the levels of growth factors, cytokines and chemokines secreted in IVM medium.
Acesso aberto (Open Access)
Estudo da combinação de preparações enzimáticas no rendimento de extração aquosa do óleo da polpa de Euterpe oleracea Martius
(Universidade Federal do Pará, 2016-03-23) FERREIRA, Erika de Souza; HERMAN, Christelle Anne Nicole Paule; http://lattes.cnpq.br/8386417434654533
The oil of the açai palm tree (Euterpe oleracea) fruits has a beneficial lipid profile for health with a high content of mono and polyunsaturated fatty acids, besides antioxidant substances, which provides its appreciation on the industry. The aqueous extraction of vegetable oils supported by enzymes has been widely used because of its benefits compared to the traditional processes. In this context, this work aims to identify the best combination of enzymatic preparations, which would maximize the aqueous extraction yield of the oil of the açai palm tree fruits from the pulp. The açai pulp was obtained from fruits collected in Abaetetuba during the 2015 crop. Four commercial enzymatic preparations (Novozymes) were used: Celluclast 1,5L (cellulase); Viscozyme L (endo 1,3(4) β-glucanase, xylanase, cellulase and hemicellulase); Ultrazym AFP (pectin lyase, cellulase and polygalacturonase); and Shearzyme 500L (endo 1,4-xylanase). The tests of açai oil extraction were performed by a process developed in this work with the following parameters: mean granulometry of the açai pulp of 0,75 cm; global dry matter of the aqueous mixtures of the pulp of 15%; individual enzymatic concentration of 1% (w:w) on pulp basis; constant orbital agitation of 100 rpm and vigorous agitation by hand every 30 minutes; incubation time and temperature of 4 hours and 50°C, respectively. The enzymatic preparations were tested individually (1x1) in quadruplicate, or combined (2x2; 3x3 and 4x4) in triplicate, totaling 89 tests. The negative control presented a mean yield of extraction of 34,91±11,81%, the lowest of all the tests. The tests performed with the isolated enzymatic preparations have enabled an increase of the extraction yield up to 36,66% compared to the negative control, confirming the important role of the enzymes in the aqueous extraction process. The tests performed with the enzymatic preparations combined 3x3 and 4x4 have increased the extraction yield up to 99,05% compared to the negative control, demonstrating the importance of using the combinations of enzymatic preparations. Among the different tests performed, the enzymatic preparations that stood out are Celluclast 1,5 L, Viscozyme L and Ultrazym AFP by facilitating the oil recovery with a reduce standard deviation (±3,01%). Furthermore, the extraction yield using this combination of three enzymatic preparations is statistically equal to the extraction yield using the four enzymatic preparations, which becomes interesting to the industry due to the cost reduction in the process.
Acesso aberto (Open Access)
Estudo de genômica comparativa de corynebacterium pseudotuberculosis linhagem 226 (biovar ovis)
(Universidade Federal do Pará, 2015-03-10) DIAS, Larissa Maranhão; CARNEIRO, Adriana Ribeiro; http://lattes.cnpq.br/7533716053525477; http://lattes.cnpq.br/7533716053525477
Corynebacterium pseudotuberculosis is a gram-positive, facultative intracellular, non-sporulating and non-encapsulated bacterium, it is non-motile although it has fimbriae, and can assume coccoid or filamentous forms (pleomorphic). Its optimum growth temperature is 37°C. This pathogen has two biovars: ovis, which usually affects small ruminants and causes caseous lymphadenitis, and biovar equi, more common in equines, bovines, camelids and bubalines, causing ulcerative lymphangitis. Its infection can lead to carcass condemnation and reduction in wool production (in ovines and caprines), milk production and meat production and, consequently, economic losses for the agricultural industry worldwide. Currently there is no effective vaccine against those illnesses. To obtain a better understanding of these species biologically, the main objective of this work is to analyze, using comparative genomics, the strain C. pseudotuberculosis 226 biovar ovis, isolated from a caprine in California, comparing it to other strains from biovars ovis and equi. The synteny analysis revealed highly conserved gene order between strain 226 and other biovar ovis strains. Phylogenomic analyses showed that the strains I19 and 267 are, respectively, the closest and the more distant phylogenetically from strain 226. Among biovar equi strains, the one with the greater phylogenomic proximity to strain 226 was strain 1/06-A. Eight pathogenicity islands were predicted, with C. pseudotuberculosis best characterized virulence genes in literature being present in island 1. No new regions related to virulence genes could be found compared to other strains. 248 orthologous genes could be found between strains I19, 267 and 226, while 282 orthologous genes could be found between strains 258, 1/06-A and 226. Based in this study it is possible to assume that strains from biovar ovis have a little varied gene repertory and strains from biovar equi have less genes shared with strain 226, reinforcing the genetic diversity between these biovars.
Acesso aberto (Open Access)
Estudo do crescimento bacteriano na presença de óleos essenciais de Dysphania ambrosioides l. e Ocimum campechianum mill. para avaliar seus potenciais como antissépticos bucais
(Universidade Federal do Pará, 2015-08-31) MOURA, Luiziana Barbosa; SANTOS, Alberdan Silva; http://lattes.cnpq.br/5976702134131016
The aromatic plants like Dysphania ambrosioides (mastruz) and Ocimum campechianum (Alfavaca), that are part of folk medicine in Brazil, have phenylpropanoids and terpenoids compounds in their essential oils as result of secondary metabolism that influence the adaptation and defense of these species in environment; particularly, defense against microrganisms. The objective of this study was to evaluate the antimicrobial activity of essential oils of D. ambrosioides and O. campechianum against pathogenic bacteria mouth. For this they used the disk diffusion method on agar and broth microdilution adapted. The plants were obtained in Santa Izabel do Pará, their leaves were washed and weighed; the essential oil was obtained by hydrodistillation. After, the components of essential oils were identified by gas chromatography-mass spectrometry. Bacteria strains used were: Lactobacillus casei (ATCC 7469), Lactobacillus fermentum (ATCC 9338), Streptococcus mutans (ATCC 25175), Streptococcus oralis (ATCC 10557) and Agregatibacter actinomycetencomitans (ATCC 29522). Bacterial suspensions were prepared for testing and 0.5 McFarland standard. As a positive control we adopted the digluconate of chlorhexidine 0.12%. Inhibition halos were observed for all samples in different concentrations of each essential oil (1%, 5%, 10%, 25%, 50% and 75%). The largest halos were found to A. actinomycetencomitans. In the microdilution test the basil oil inhibited the bacteria S. mutans, the main etiological factor for caries in concentration of 1%; mastruz the oil inhibited L. casei, microorganism that enhances the process of decay in concentrations from 10%; Both oils inhibited the growth of A. actinomycetemcomitans, and can be effective against periodontal disease caused by that pathogen. Plant species of this study produce secondary metabolites classes with potential applications in the development of dental products.
Acesso aberto (Open Access)
Identificação de polimorfismos de nucleotídeo único com efeitos deletérios em transcriptomas de câncer gástrico
(Universidade Federal do Pará, 2015-09-29) SILVA, Viviane Santos da; DARNET, Sylvain Henri; http://lattes.cnpq.br/4586614214029929
Gastric cancer is one of the leading causes of cancer mortality in the world. Its development is associated with factors related to lifestyle and genetic alterations that can modify genes and important biosynthetic and metabolic pathways that help maintaining cellular and tissue integrity. One of the main cancer research objectives is identify genes and mutations that may be used as genetic markers and potential therapeutic targets. Studies with genes related to the pathway of steroid hormones proteins have already identified polymorphisms that may be related to the risk of cancer. Because it is an important route for physiological and pathological processes identification of genetic polymorphisms in this and cholesterol biosynthesis pathway may contribute to the understanding of gastric carcinogenesis. Therefore, we performed a comprehensive bioinformatics analysis of transcriptome datasets from gastric tissues with and without cancer, in order to identify SNPs in genes associated to enzymes of biosynthetic pathways of steroid hormones, cholesterol and progesterone receptor that may be related to gastric cancer. The analysis was performed using the Galaxy Platform, the Bowtie alignment tool and the Provean software for functional analysis of variations. Deleterious mutations have been identified in CYP51A1, DHCR24 and SQLE genes in the steroid hormone biosynthesis pathway, and in CYP3A5, HSD17B12, UGT1A1, UGT1A5, UGT1A6 and AKR1C3 genes in the cholesterol biosynthetic pathway. The CYP3A5 gene had the highest number of deleterious SNPs. These results indicate a possible participation of genes analyzed in the molecular mechanisms involved in the development of gastric cancer.
Acesso aberto (Open Access)
Influência dos metabólitos secundários de Piper divaricatum da região amazônica no controle do Fusarium solani f. sp. piperis causador da fusariose em pimenta do reino
(Universidade Federal do Pará, 2014-09-18) MEIRELES, Erisléia das Neves; RAMOS, Alessandra de Rezende; http://lattes.cnpq.br/1279694874191138; SILVA, Joyce Kelly do Rosário da; http://lattes.cnpq.br/2278686174214080
The culture of black pepper (Piper nigrum L.) is one of the main agricultural activities in Pará State and it suffers serious damages caused by fusariosis, disease restrict in Brazil. This paper evaluated the antifungal activity in vitro of essential oils of species of Piper rich in phenylpropanoid before the Fusarium solani f. sp. piperis, which causes the fusariosis. The inhibition of mycelial growth by the method of agar dissemination in concentration of 5 mg.mL-1 was considered: decrease to P. aduncum (20.3%) and P. Krukoffii (31.4%); moderated to P. callosum (55.7%) and P. marginatum (70.3%) and high to P. divaricatum (93.3%). The major components identified by CG-EM were dilapiol (92.0%), safrole (78.0%), methyleugenol (75.2%) and eugenol (7.9%), apiol (80.0%), Z-isoosmorizol (44.0%) and E-anethole (22.0%), respectively. The oil of P. divaricatum and its major compounds present CIM values of 0.75 mg.mL-1. The evaluation of the effects combined of eugenol and methyl eugenol pointed eugenol as the main responsible by the activity.
Acesso aberto (Open Access)
Método de identificação de genes taxonomicamente restritos em dados de RNA-seq em organismo não modelo
(Universidade Federal do Pará, 2015-09-28) OLIVEIRA, Lorena Silva de; DARNET, Sylvain Henri; http://lattes.cnpq.br/4586614214029929
Black pepper (Piper nigrum) is a biotechnologically interesting plant, research target for both metabolism exploration and improvement related to phytopathological problems, in addition to understanding the evolution of basal angiosperms, ancestral group to which it belongs. With the technological revolution, the next generation sequencing offered access to genetic heritage of non model plants enabling the opening of new biotechnological perspective. The identification of non homologous genes restricted to certain species, called taxonomically restricted genes (TRGs), is a primary biotechnological target, especially in species and groups that are divergent and ancestral. This study aims to establish a method for TRGs identification from RNA-seq data and to validate the approach a dataset for black pepper. The method consists in filtering the transcripts in several stages, so that the annotated transcripts and false positives are removed, and the remaining data without molecular information are classified as potential TRGs. The application of this approach to a black pepper transcriptome dataset (35,631 transcripts) resulted in 22,661 transcripts annotated by similarity. The transcripts that were not annotated in this first analysis were processed in the TRAPID tool, resulting in 12,895 transcripts not annotated. The evaluation of transcripts for false positive detection resulted in 245 true transcripts that were analyzed for the presence of non-coding RNA, resulting in 204 unidentified transcripts. At the end of the method application 71 non annotated transcripts remained with coding regions of protein, indicating potential TRGs. The characterization of these potential TRGs in black pepper can provide new information about the molecular mechanism of this specie and perhaps elucidate pathways for the establishment of cultivars tolerant to disease.