01.1 - Graduação (Sede)
URI permanente desta comunidadehttps://arandu.ufrpe.br/handle/123456789/2
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Resultados da Pesquisa
Item Avaliação do fluxo sanguíneo do folículo ovulatório e corpo lúteo de fêmeas bubalinas por ultrassonografia Doppler(2024-09-27) Farias Júnior, Francisco Timbó; Batista, André Mariano; http://lattes.cnpq.br/5615914349535394; http://lattes.cnpq.br/7562129132084802The Mandatory Supervised Internship (ESO) is an essential part of the training of a Veterinary Doctor, being a mandatory requirement in the Veterinary Medicine course at UFRPE. With a workload of 420 hours, the internship was conducted at the Laboratory of Biotechniques Applied to Reproduction (LBR), between 04/01/2024 and 06/14/2024. The focus was on improving reproductive biotechnology, with emphasis on the in vitro maturation of oocytes and the development of an article based on research carried out with Murrah buffaloes. The study evaluated the blood flow of the ovulatory follicle and corpus luteum using Doppler ultrasound, correlating these data with reproductive efficiency. In the laboratory, activities such as in vitro maturation of bovine oocytes (IVM) and evaluation of meiotic progression using epifluorescence microscopy were carried out. Under the guidance of Professor Dr. André Mariano Batista and supervision of Professor Dr. Cláudio Coutinho Bartolomeu, the internship provided a practical insights into reproductive biotechnologies and their impact on reproductive efficiency in buffaloes.Item Relatório de Estágio Supervisionado Obrigatório (ESO) realizado na Empresa Brasileira de Pesquisa Agropecuária - EMBRAPA Gado de Leite, Juiz de Fora e no Laboratório de Biotécnicas Aplicadas à Reprodução do Departamento de Medicina Veterinária da Universidade Federal Rural de Pernambuco: efeito da adição de nanoemulsão à base de óleo essencial rico em limoneno ao diluente de refrigeração de sêmen caprino(2024-02-15) Nottar, Luiz Henrique Todescatt; Batista, André Mariano; http://lattes.cnpq.br/5615914349535394; http://lattes.cnpq.br/3747311074315408The present study aims to report the activities carried out in compliance with the discipline 08525 - Mandatory Supervised Internship (MSI), part of the Bachelor's Degree in Veterinary Medicine, conducted between October 2, 2023, and November 28, 2023, at the Brazilian Agricultural Research Corporation (Embrapa), in the Dairy Cattle unit in the municipality of Juiz de Fora - MG, and between January 2 and January 19, 2024, at the Laboratory of Biotechniques Applied to Reproduction (LBR) of the Department of Veterinary Medicine (DMV) of the Federal Rural University of Pernambuco (UFRPE). The activities carried out provided in-depth training in in vitro embryo production (IVP) techniques for research purposes, including follicular aspiration of ovaries from abattoirs, in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro embryo culture (IVC). Activities in the field of molecular biology involving DNA extraction and amplification through the Polymerase Chain Reaction (PCR) technique were also performed. In the second chapter of this study, a research was conducted to evaluate the effect of adding nanoemulsion based on limonene-rich essential oil extracted from wild orange peel (NELS) to the semen extender of goat breeders. Seminal samples from five Saanen breed goats were used for the research. Each seminal pool was diluted in Tris-egg yolk supplemented with NELS at concentrations of 1.25, 2.5, and 5% at a concentration of 200 × 106 sperm/mL. The samples were placed in Eppendorf tubes (1.5 mL) and stored in refrigeration boxes (botuflex®) for cooling curve determination up to 5 ºC, and after 2 hours, they were transferred to the refrigerator, also at a controlled temperature of 5 ºC. For each analysis, part of each sample was aliquoted and transferred to other Eppendorf tubes (1.5 mL) in a dry bath at 37 ºC and subsequently evaluated in the Computer-Assisted Sperm Analysis System (CASA) for sperm kinetics, and in epifluorescence microscopy for Mitochondrial Membrane Potential (MMP), Plasma Membrane Integrity (PMI), and Acrosomal Membrane Integrity (AMI). The addition of wild orange nanoemulsion to the Tris-egg yolk extender was not able to improve sperm motility and kinetic parameters, besides significantly reducing MMP after 24 hours of refrigeration.Item Caracterização de leveduras presentes na microbiota do melão amarelo (Cucumis melo L.), quanto a capacidade de produção e manutenção de biofilme(2021-02-09) Ramos, Sérgio Batista; Laranjeira, Delson; Coelho, Iwanne Lima; http://lattes.cnpq.br/0302261951754304; http://lattes.cnpq.br/1262204427401043; http://lattes.cnpq.br/4051133932371862Item Caracterização eletroquímica das interaçôes da lectina de sementes de Cratylia mollis (Cramoll) com glicose e eritrócitos de coelho em biossensor de MOF de [Cu3(BTC)2.(H2O)3]n(2022-05-31) Carvalho, Maryana Hermínio de; Coelho, Luana Cassandra Breitenbach Barroso; Freitas, Kátia Cristina Silva de; http://lattes.cnpq.br/5859266863241551; http://lattes.cnpq.br/2944428818449047; http://lattes.cnpq.br/0134945103585503Biosensors are small devices known for their selectivity, high specificity and sensitivity with the respective analyte even at low concentrations. Thus, we aimed to evaluate the charge transfer and electrochemical potential of the plant popularly known as camaratu bean, a forage native to the Caatinga biome, rich in shrub legumes, from which Cramoll lectin is extracted from Cratylia mollis Mart. electrochemical biosensor using the crystalline polymer MOF- [Cu3(BTC)2.(H2O)3]n.. to immobilize and characterize the physical and chemical properties of C. mollis (Cramoll) seed lectin and its interaction with free carbohydrates (glu-cose) and carbohydrates on the surface of rabbit erythrocytes. Lectin immobilization in MOF was effective for the construction of the electrochemical system, showing itself to be sensi-tive to conformational changes, being able to detect changes in the electrochemical poten-tials of Cramoll promoted in a buffered medium (200 mM potassium phosphate buffer, pH 7.0) with different glucose concentrations, highlighting the concentrations of 10 mM (96 - 121 mV), 15 mM (110 - 126 mV) and 20 mM (107 - 142 mV) in the time of 5 to 30 min. In the cyclic voltammetry, the interaction in the potential range from -0.2 to 0.0 V stood out; with the current reaching 0.0003 A and at peak 2 in the potential range of 0.4 to 0.5 V; with the current reaching 0.0007 A. The interaction of the carbohydrate present on the surface of the rabbit erythrocyte was also monitored by the potentiometric method, detecting changes in the Cramoll-glucose potentials, even in the presence of rabbit erythrocytes. Through SEM, it was possible to observe Cramoll immobilized on the MOF surface, proving the specificity of the ligand (glucose-lectin) through the morphological changes of this protein in this process. This electrochemical model, Cramoll/MOF biosensor, is effective for evalu-ating free lectin/carbohydrate or in the erythrocyte membrane.Item CRISPR-Cas9 no setor florestal: uma abordagem cienciométrica(2021-12-09) Braz, Luísa Silva; Gallo, Ricardo; http://lattes.cnpq.br/5160912065817980; http://lattes.cnpq.br/2573015676942351CRISPR-Cas9 is a genetic editing technique that works like a pair of scissors, adding new information to the DNA after “cutting” it into a small, previously programmed part. This technological innovation brings great potential for the genetic improvement of plants, as it has a low cost and is simple to implement, when compared to other biotechnological techniques. Therefore, this work aimed to carry out a scientometric survey of research on CRISPR-Cas9 applied in the forest sector. Using the bibliometric method, the term “CRISPR-Cas9” combined with the following words was searched in the Web of Science and Scopus databases: "plant breeding", "tree", "forestry", "tree breeding"; “forest”. The articles obtained were analyzed on bibliometrix, generating data for scientometric analysis. A huge growth in the number of publications on the topic was observed, as of 2014. It was also found that 10 journals form the core of publications on the topic, according to Bradford's Law; and that the main publishers are: International Journal of Molecular Science, Frontiers in Plant Science and Plant Biotechnology Journal. The magazines with the greatest impact were: Plant Biotechnology Journal and Nature Communications. The most prominent authors were: Zhang Y, with 24 articles published, Li Y, with 19, Wang Y, with 17, Wang X, with 15 and Liu Y, with 11. In the analysis of Lotka's Law, we could see that 79 .3% of authors wrote only 1 article each on this topic, while few authors had a high number of publications. The survey also provided the researchers' affiliations. The University of California, South China Agricultural University, and University of Chinese Academy of Sciences are standind out with a high number of publications. The theme is being researched mainly in institutions in the United States and Asia, with research being carried out in Brazil. The words (Keywords Plus): "gene editing", "genetics", "plant breeding", "article" and "nonhuman" were the most cited in the articles, as for the keywords of the authors of the articles: tomato, potato, rice , Populus and Bombyx mori L., presented high frequency. Research involving CRISPR-Cas9 in forestry production is still in its beginning, noting that species of the genus Populus stood out in several studies, however it is possible to observe a considerable advance for agricultural crops, such as rice, tomatoes and potatoes. Forest species are still in the initial stages of using this technique, but they have great potential for improvement in clones and hybrids of species of commercial interest.Item Ciências “ômicas” aplicadas à análise de sementes(2020-11-05) Santana, Bárbara Jossany Gomes de; Santos, Marcone Moreira; Silva, Joselane Príscila Gomes da; http://lattes.cnpq.br/8028063388524508; http://lattes.cnpq.br/3284707164193715; http://lattes.cnpq.br/7128880355985474Seed analysis consists of the evaluation of seed quality through the use of different techniques for the most varied purposes. Many tools can be used, among them the “omics” sciences. This set of approaches can be considered a substantial technological advance for seed analysis. Within the techniques that are part of this set of science, the following stand out: transcriptomics (study of genetic transcripts, RNA‟s), proteomics (study of proteins), metabolomics (study of metabolites) and lipidomics (study of lipids). However, in relation to the forest seed analysis, these techniques are still not widespread. Thus, the objective of this work was to develop an exploratory study on the use of “omics” sciences for seed analysis, highlighting their objectives and technologies used. To this end, a database was created using Google academic as a search tool. Search terms were established in Portuguese and in English, which were: transcriptoma, proteoma, metaboloma, lipidoma para análise de sementes e para sementes florestais, transcriptomic, proteomic, metabolomic, lipidomic for seeds, for forest seeds analysis, for seeds analysis. The selected studies were grouped in tables according to the technique used: Transcriptomics, Proteomics, Metabolomics and Lipidomics. Species, purpose of the study, classification regarding use (Agricultural, Forestry, Medicinal and Others), origin of the studies (national or international) and year of publication were also identified. A total of 105 studies were analyzed, in which 64 corresponded to agricultural species, 18 forestry, 13 others and 10 were classified as medicinal. Between all the studies analyzed, 77% corresponded to international studies. Among the national studies, the most expressive techniques were: Transcriptomics and Proteomics with 38% and 28%, respectively. Regarding the year of publication, the oldest was Proteomics, in 2001. Regarding the classification of use for the species studied, the largest number of studies was developed with agricultural species, due to the greater development and investments of the sector, which has more resources for investment in advanced technologies. However, even if forest species are less studied, it is observed that the field of omics is a promising and developing area, and may have several applications in the future. Therefore, the importance of expanding investments in research in the field of forest seeds is emphasized, especially for native species, whose knowledge is incipient in view of the diversity and importance of species.Item Potencial biotecnológico do inibidor de tripsina de flores de Moringa oleifera: uma revisão(2021-12-16) Medeiros, Êsdras da Silva; Pontual, Emmanuel Viana; http://lattes.cnpq.br/1777060469196142; http://lattes.cnpq.br/8379001129790219This work provides a review on the applications of the Moringa oleifera flower trypsin inhibitor (MoFTI) as an insecticidal, antiparasitic, antitumor, immunomodulatory and anti-inflammatory agent. MoFTI is a 18.2-kDa protein (Ki against trypsin of 2.4 μM) that killed (LC50 of 0.3 mg/mL) or delayed the development of newly hatched Aedes aegypti larvae and impaired the growth and viability of the larval gut microbiota (MIC of 0.031 mg/mL; MBC of 1.0 mg/mL). It was also reported that MoFTI caused lysis of Trypanosoma cruzi trypomastigotes (LC50 of 41.2 μg/mL). The inhibitor stimulated the release of the inflammatory cytokines TNF-α and INF-γ, the anti-inflammatory cytokine IL-10, and NO by T. cruzi-infected human peripheral blood mononuclear cells (PBMCs) with the advantage of exerts low toxicity to murine peritoneal macrophages and Vero cells, and no toxicity to human PBMCs. It is known that MoFTI antitumor activity involves the reduction of sarcoma 180 tumor weight in mice and the impairment of the vascularization in tumor microenvironment without exerting toxicity to the animals. Also the inhibitor is an anti-inflammatory and immunomodulatory agent because it reduced paddle edema and leukocyte migration to the peritoneal cavity in mice, and modulated levels of NO, pro-inflammatory (TNF-α, IL-6 and IL-17A) and anti-inflammatory (IL-4 and IL-10) cytokines, respectively. The reviewed data implicate MoFTI as an interesting molecule for biotechnological applications, whether due to its insecticidal potential or pharmacological value.Item Produção de proteases por Aspergillus ochraceus URM 604 obtidas por fermentação em estado sólido utilizando farelo de trigo e resíduo de café como substrato(2021-03-03) Santos, Amanda Lucena dos; Porto, Ana Lúcia Figueiredo; Cardoso, Kethylen Barbara Barbosa; http://lattes.cnpq.br/4784303425329040; http://lattes.cnpq.br/4989617783837981; http://lattes.cnpq.br/3697084385877618Proteases are enzymes of great commercial interest, since it has several industrial applications, such as the pharmaceutical, food, beverages and cleaning products. Among the organisms capable of producing these enzymes are filamentous fungi, having as advantages the possibility of secreting enzymes in the extracellular medium and growing on low-cost substrates. The Solid State Fermentation (SSF) is one of the recommended techniques in the cultivation of filamentous fungi, especially because it simulates its natural habitat, favoring their growth. Aiming the importance of proteases and the growing demand of the global market needs, it is necessary to search for new sources and better production methods. Thus, the objective of this research was to produce proteases by the filamentous fungus Aspergillus ochraceus URM 604 under SSF using substrates derived from the agribusiness, in 2³ factorial design. It was investigated the influence of the type of substrate (coffee residue, wheat bran and 1: 1 coffee + wheat bran), amount of substrate (3g, 5g and 7g ) and humidity (20, 40 and 60%) to determine the ideal conditions for protease production. Fermentation took place for 7 days at 30 °C and the metabolic extract was used for further analysis. For biochemical and physical analysis the protein activity, total proteins, temperature and optimal pH of the obtained enzyme were determined. When analyzing the influence of the variables adopted in the 2³ factorial design, only the type of substrate was a significant parameter. The best substrate was wheat bran, which showed a specific enzymatic activity of 218.27 U / mg under 3g of substrate and 60% humidity. The other conditions also showed high results when compared to the literature. The optimum temperature of the enzyme produced was 50°C and the optimum pH was pH 8-9 (alkaline protease). Thus, this research shows that the fungus A. ochraceus URM 604 has biotechnological potential for protease production under SSF using low-cost substrates such as coffee grounds and wheat bran, this being the first report for the species.Item Recuperação e purificação parcial de proteases colagenolíticas de tainha (Mugil liza) usando precipitação e particionamento em sistemas de fases(2020-02-03) Costa, Beatriz de Aquino Marques da; Porto, Ana Lúcia Figueiredo; http://lattes.cnpq.br/4989617783837981; http://lattes.cnpq.br/2650705679652460Proteases play an important role in the field of biotechnology studies, which is why the research for alternative sources is highly desirable. Thus, combining extraction, recovery and purification techniques to alternative sources, such as by-products of the fishery production chain, in order to generate the least possible damage to the aquatic environment is profitable for the global enzyme market. Therefore, the objective of the present work is to partially recover and purify collagenolytic proteases from Mullet (Mugil liza) digestive viscera with potential biotechnological application. For this, the digestive viscera extract (intestine, liverand a mixture of various viscera) was precipitated using ammonium sulfate ((NH4) 2SO4). In addition, the intestinal viscera were submitted to partitioning by an Aqueous Two-Phase System (ATPS) and a Three-Phase Partitioning system (TPP), to evaluate the most beneficial conditions for the purification of collagenolytic proteases. The results obtained demonstrate that, for ammonium sulphate precipitation, the best results of Purification Factor occurred in concentrations of 30-60% for all assessed extracts (intestine, liver and mix); the aqueous two-phase system (PEG/Citrate) carried out with Mullet intestinal viscera extract demonstrates that the conditions: PEG of 8000 g/mol molar mass; 20% concentration of PEG; 15% citrate concentration lead to the highest purification factor. It is also observed that the collagenases tend to migrate to the PEG-rich phase; for the three-phase system (t-butanol / (NH4) 2SO4) the highest recovery rate of collagenolytic protease was observed in the proportion of 1:0.5, obtaining recovery of 240.01% and a purification factorof 2,55. Thus, it is concluded that the tests can be used in the recovery of biomolecules from neglected organic solid waste from the fishing industry.Item Produção, extração, purificação e caracterização de proteases fibrinolíticas produzidas por Streptomyces parvulus DPUA 1573(2021-11-29) Nascimento, Maria Clara do; Bezerra, Raquel Pedrosa; Batista, Juanize Matias da Silva; http://lattes.cnpq.br/6699725036732885; http://lattes.cnpq.br/1466206759539320; http://lattes.cnpq.br/5929405825655717Due to their fibrin degradation potential, fibrinolytic proteases are a promising alternative in the pharmaceutical industry for the treatment of cardiovascular diseases, especially thrombosis. There are several sources of fibrinolytic proteases, however, the microbial sources are the ones that stand out in terms of low cost and high production rates. From their production to application, enzymes need to go through several processes, which sounds negative, making the steps more costly and late. A method capable of overcoming these problems is the aqueous two-phase system (SDFA), a process capable of reducing downstream steps. The objective of this work was to produce, purify and biochemically characterize the fibrinolytic protease produced by Streptomyces parvulus DPUA 1573. The protease was produced by submerged fermentation using agro-industrial waste or co-products. The crude extract that showed the highest enzymatic activity (passion fruit peel flour) was subjected to extraction by SDFA consisting of polyethylene glycol (PEG) and phosphate salts (potassium and sodium), following a 24 plan. After extraction by SDFA, the protease was subjected to purification by gel filtration chromatography, and already purified had its biochemical characterization performed. The protease produced by S. parvulus DPUA 1573 showed fibrinolytic activity of 15.46 U/mL and was able to form a halo of 317.31 mm2 acting on fibrin degradation. In SDFA, the fibrinolytic protease partitioned preferentially to the PEG-rich phase. The best assay selected according to the combination of the highest specific activity index, purification factor and activity yield was 16, composed of PEG 8,000 gmol-1, 17.5 v/v PEG, pH 8.0 and 15 v/v of phosphate salts. The protease activity of the enzyme was highly stimulated in the presence of iron, reaching a 55% increase in activity, and drastically decreased in the presence of the protease inhibitor 2-mercaptoethanol (91%). The optimum temperature and pH for the enzymatic activity were 40ºC and pH 7.0, respectively, keeping the enzyme activity stable between 30ºC and 60ºC and in the pH range from 7.0 to 8.5. Based on the analyzed results, it was seen that S. parvulus DPUA 1573 proved to be a good producer of fibrinolytic proteases, and the PEG/Phosphate aqueous two-phase system proved to be a great alternative for the extraction and pre-purification of fibrinolytic proteases.