DSpace Communidade: Programa de Pós-Graduação em Biotecnologia Aplicada à Agropecuária

Vitrificação em fluido folicular bovino.

2026-01-01T00:00:00Z

Título: Vitrificação em fluido folicular bovino. Autor(es): MERCÊS, Marcela Oliveira das Abstract: Vitrification is one of the methods used for immature and mature bovine oocytes. However, it needs improvement regarding hearing competence after warming. The use of bovine follicular fluid (BFF) is known in in vitro production systems, but unexplored in vitrification media. The aim of this study was to vitrify oocytes in a solution supplemented with bovine follicular fluid (BFF) or fetal bovine serum (FBS) to investigate post-warming survival through nuclear maturation and analysis of HSP70 gene expression. The study was divided into two experiments: In experiment 1, immature cumulus oophorus complexes (COCs) were distributed into three experimental groups. In experiment 2, three experimental groups were used for mature oocytes and three experimental groups for immature cumulus oophorus complexes. The control group of oocytes (without vitrification) in immature oocytes were directly subjected to in vitro maturation. Both experiments used, in group 1, (TCM 199 medium + 20% fetal bovine serum (FBS) + cryoprotectants), and in group 2, vitrification in (TCM 199 medium + 20% bovine follicular fluid (BFF) + cryoprotectants), followed by in vitro maturation of immature oocyte complexes (OCCs). In experiment 1, all immature oocytes from the experimental groups, after maturation, were stained with acetic orcein for structural visualization. Meanwhile, in experiment 2, matured and immature oocytes, after warming, were subjected to gene expression analysis of the heat shock protein HSP70. Survival rates of oocytes that were matured to assess meiotic stage II (MII) were expressed as percentages and evaluated by analysis of variance (ANOVA) and Bonferroni post-hoc test. For statistical analysis, SigmaPlot® 14.5 software was used with a significance level of P<0.005. The statistical analysis of gene expression was performed using two-way ANOVA in the Sigmaplot program with a significance level of 0.05. In the maturation analysis of immature oocytes, no significant differences were found between the groups vitrified with FBS or FFB (p>0.001), and the maturation rate was lower (p>0.001) than the control group. Regarding experiment 2, oocytes from the vitrified matured group, the gene expression analysis of the HP70 protein showed no difference between the groups vitrified with FBS and FFB, while the granulosa cells of immature oocytes, FFB, and FBS also differed from each other and from the control (p < 0.05). These results indicate that follicular fluid can be used to replace FBS supplementation in vitrification media of bovine oocytes.

Montagem, predição gênica e anotação do genoma da Cynoscion acoupa (Lacepède, 1801).

2025-07-14T00:00:00Z

Título: Montagem, predição gênica e anotação do genoma da Cynoscion acoupa (Lacepède, 1801). Autor(es): OLIVEIRA, Rosyely da Silva Abstract: Aquaculture in Brazil had positive results in 2023, where national production reached 887,03 tons. The Cynoscion acoupa is a fish popularly known as “pescada-amarela”, found mainly on the north coast and has a great economic value mainly for gastronomic purposes. More recently, a material called "isinglass" was observed, found in the swim bladder of this fish, which has been used as raw material in several areas of the market, such as the pharmaceutical, civil, food, among others. The purpose of this work is to perform the genome assembling, gene prediction and carry out functional annotation of the C. acoupa organism. DNA samples were extracted from 5 specimens in the regions of Salinópolis-PA, Bragança-PA and on the coast of Amapá. The best sample was chosen to continue the study. Through these samples, the complete genome of the captured individuals was sequenced, with two paired libraries paired-end short insert (2 x 250 bp) DNAseq built with the Illumina DNA prep Kit, using the NovaSeq SP 6000 Illumina sequencing platform, producing raw data. Subsequently, FASTQC were used to verify the quality of the sequencing. From the generated sequences the de novo genome assembly will be made using the ABYSS assembler. BOWTIE-2 will be used to align the contigs and scaffolds with the genome of a phylogenetically close species, GeneMark will be used for gene prediction and BUSCO for genome annotation, so that it can finally be deposited at the NCBI. Through the results, it is expected to bring new information about genome sequencing and its quality, together with the publication of these data for future studies.

Análise da expressão de miRNAS e genes-alvo em sangue periférico de equinos quarto de milha antes e após treino.

2025-01-30T00:00:00Z

Título: Análise da expressão de miRNAS e genes-alvo em sangue periférico de equinos quarto de milha antes e após treino. Autor(es): AMARAL, Thiago Maués Abstract: In the world, there are approximately 60 million horses, Brazil is the 4th country with the largest number of horses in heads (≤6 million). Brazilian Agribusiness generates around R$ 7.5 billion annually and the use of horses in sports is the most profitable segment of equinoculture. In chapter I, the objective was to carry out a literature review, seeking to understand the use of the Quarter Horse and its main related activities, as well as to understand the animal physiology related to exercise. For review, 7 base articles and 92 articles for discussion were used. In chapter II, the objective of the research was to investigate the differential expression of microRNAs and participatory target genes in exercise metabolic pathways, and to verify hematological and clinical alterations in QM horses under different physiological conditions. For the experiment, 10 horses (Equus caballus) of the quarter mile breed were used. The animals were submitted to physical efforts with obstacle avoidance and were clinically evaluated before and after exercise. Respiratory Rate, Heart Rate and Rectal Temperature were used as clinical parameters, 19 whole blood parameters were used as hematological parameters (complete blood count), for molecular evaluation the ALDOA and EGFR genes and the miR-133b and miR-1260b microRNAs were selected. . As clinical results, there were statistical differences for FR (<0.0001), FC (<0.0001) and TR (<0.0001), these parameters should be considered as important in the management of animal health, as well as evaluating the physiological performance of horses during training. The 9 hematological parameters that showed significance (p<0.05) before and after exercise were: Erythrocyte; hematocrit; Hemoglobin; Mean Corpuscular Volume; Range of distribution of erythrocytes; Mean Corpuscular Hemoglobin, Neutrophils and Lymphocytes. These parameters are related to aerobic capacity due to increased oxygen transport, making equine health indicators. The expression of genes ALDOA (p<0.0454) and EGFR (p<0.0198) and microsRNAs -133b (p<0.0240), -1260b (p<0.0066) were significant after exercise. The regulation of the ALDOA gene is associated with the metabolism of oxygen for the conversion of energy, the regulation of the EGFR may be more related to the post-training recovery provoked by the inflammatory response responsive to the exercise, due to the great physiological stress and energy expenditure, the miR -133b and -1260b had not yet been reported in the literature associated with modulation of exercise metabolic pathways, despite not regulating the genes of this research, these micros showed to be regulated after exercise, which shows that they regulate other target genes. These biomarkers found can be used as a reference for athletes of the QM race created in the north and northeast of Brazil. The changes observed for some parameters in the variables weight and sex had no clinical relevance (p>0.05). A future biochemical examination may show evidence of other biomarkers.

Potencial biotecnológico do uso de filtrados de Trichoderma asperellum com indução de quitinases e proteases no controle de larvas de Tenebrio molitor (Coleoptera: Tenebrionidae).

2024-09-09T00:00:00Z

Título: Potencial biotecnológico do uso de filtrados de Trichoderma asperellum com indução de quitinases e proteases no controle de larvas de Tenebrio molitor (Coleoptera: Tenebrionidae). Autor(es): FIGUEIREDO, Mayara Pimentel Abstract: Trichoderma sp. it is widely used for controlling various fungi that cause plant diseases, but few studies report their insecticide action. The Trichoderma asperellum species although present in many tropical soils, is still studied with low frequency. Six isolates of T. asperellum the Brazilian Amazon were selected after toxicology screening and evaluated for insecticidal activity against larvae of the beetle Tenebrio molitor, insects are often used as a reference laboratory for testing with biological agents. In this work we used the entomopathogenic fungus Beauveria bassina as positive control and distilled water as negative control in comparison to isolates of T. asperellum. Analyzed spore suspensions at a concentration of 108 spores/mL, filtered with and without enzyme induction of chitinase and protease after T. asperellum growth and B. bassiana in liquid Minimum Medium (MM), on larvae of 3º instar 7. molitor and enzymatic activity of filtered, and also the filtered were autoclaved at 120 º C temperature to see if there is action of enzymes in the middle. The isolates of 7. asperellum that stand out with chitinase containing filtrates were UFRA T192, TI66 UFRA UFRA and T57, as caused mortality of 100, 77,5 and 77,5%, respectively, differing from the positive control causing B. bassiana 50% mortality, and the negative control with sterile distilled water to 2,5%. All treatments 7. asperellum protease induction caused 100% mortality equal to the positive control with B. bassiana. The highest chitinolytic activities were observed for the T. asperellum UFRA T127, T57 UFRA, UFRA UFRA T166 and T175, with 0,21208, 0,18598, 0,18036 and 0,1704 U / ml, whereas B. bassina had lower activity 0,1536 U / mL. Most proteolytic activity obtained was isolated from 7. asperellum UFRA T175 with 0, 54214 U/mL and B. bassiana 0,10899 U/ml. The 7. molitor reduced weight after application of treatments with enzymes. The filtrates were autoclaved isolates that showed lower mortality rates when compared with non-filtered autoclavable. It is concluded that T. asperellum produce chitinases and proteases, and these enzymes are involved in insecticide similar to what occurs with the entomopathogenic fungi.