Programa de Pós-Graduação em Ciência Animal - PPGCAN/Castanhal

URI Permanente desta comunidadehttps://repositorio.ufpa.br/handle/2011/2335

O Programa de Pós-Graduação em Ciência Animal (PPGCAN) é um programa do Campus Universitário de Castanhal (CCAST) da Universidade Federal do Pará (UFPA), Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) e Universidade Federal Rural da Amazônia (UFRA). O Programa iniciou o nível de Mestrado em 1999 junto à CAPES/MEC, tendo aprovação de sua proposta de Doutorado junto à agência em 2008, com início da primeira turma em 2009, o que certamente contribuirá à formação de massa crítica regional.Se caracteriza por apresentar um conjunto de disciplinas que repassam aos alunos um embasamento teórico e prático, capacitando os mesmos a aplicarem os conhecimentos adquiridos no avanço regional da produção animal. Direcionando as linhas de pesquisa do curso à nutrição, biotecnologia, pastagens, conservação de recursos genéticos, reprodução, sanidade animal, bovinocultura, bubalinocultura, animais silvestres, ecologia aquática e aquicultura.

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Acesso aberto (Open Access)
Avaliação da dinâmica folicular, atividade ovariana e involução uterina durante o pós-parto de búfalas (Bubalus bubalis) criadas em condições tropicais
(Universidade Federal do Pará, 2015-08-11) SILVA, Aluizio Otavio Almeida da; RIBEIRO, Haroldo Francisco Lobato; http://lattes.cnpq.br/1614582293203770
In this experiment aimed to monitor the follicular dynamics and ovarian activity of buffalo females in the postpartum period through ultrasound exams, investigating whether some events related to ovarian activity in the Postpartum Period (PPP), and uterine involution, since the 7th day calved until the second post-partum estrus. ten were used (10) crossbred females between Murrah and Mediterranean breeds with eutócicas parities and no history of reproductive problems, managed in semi extensive system, with availability of water and mineral supplementation ad libitum. It was found the presence of follicular growth patterns of two waves (2 OCF) and three waves (3OCF) at a ratio of 40% and 60%, respectively. The 1st and 2nd postpartum estrus demonstrated to 37,00+13,12 and 58,83+14,56 days postpartum for 2OCF animals, 45,00 + 15,19 and 66,00 + 23.02 days postpartum for animal 3OCF. The estrous intervals were 21,00+2,44 and 21,83+2,56 days for animals 2OCF and 3OCF respectively. Follicular emergency occurred at an average of 1,02+0,34 and 7.25+0,50 days cycle for the first and second waveform, respectively, of animals and 2OCF 0,93+0,35; 6,6+1,36 and 10,83+1,94 days cycle for the first, second and third wave, respectively, for the animal 3OCF. The pre ovulatory follicular diameter was 13,08±2,6 and 11,45+0,35 mm for the 1st and 2nd postpartum estrus of 2 OCF animals and 13,24+2,41 and 12,74+1,10 mm for the 1st and 2nd postpartum estrus of 3OCF animals. The uterine involution occurred at an average of 29,00+1,48 days, (P <0.001), diameter of the post gravid uterine horn (DPGUH) around 18,84+0,33 mm. The rate of uterine involution was 0,97+0,21 mm/day, (P <0,001). There was a high correlation between the PPP and CUPG, represented by r = -0,8179 (P<0,0001), being possible to establish a linear regression between the two parameters, represented by the equation Y = 40,8809 + (-0,8214.X) (P<0,0001).
Acesso aberto (Open Access)
Avaliação da técnica de aspiração ovocitária transvaginal (Ovum Pick Up) e produção in vitro de embriões da raça nelore (Bos taurus indicus) oriundos de doadoras com alterações da fertilidade
(Universidade Federal do Pará, 2003-09-11) SOUSA, Alysson Jorge de Oliveira; OHASHI, Otávio Mitio; http://lattes.cnpq.br/5547874183666459
The objective of this experiment was evaluate the possibility of the use of zebu cattle females with raised genetic quality presenting problems of fertility, acquired after a program of conventional transference of embryos, as oocyte donors in a program of Ovum Pick Up and In Vitro embryo production. Sixteen females of Nelore breed (Bos taurus indicus) with average age of 11 years and 7 months, had been submitted to the same conditions of enviroment, feeding and management. These animals were removed of the program of embryo transfer due fertility problems (ovarian cysts, genital infections and some cases of repeat breedings). The animals had been divided in two groups in agreement the type of pathology: Group I (n=08) formed by animals that had presented ovarians patologies (cysts) and GROUP II (n=08) formed by animals with extra-ovarians patologies, both groups had been submitted to aspiration follicular (OPU) using two types of needles (21G ; 19G ) and pressures (55 e 70mmHg). The two groups were divided in four treatments (T1 =55/21G; T2 = 55/19G; T3 = 70/21G e T4 = 70/19G) and after oocites collection they were submitted to IVP. In this work, 149 aspirations were realised and 879 oocytes were collected (5,9 4,88) GRADE I = 16%; GRADE II = 43%; GRADE III = 21% e GRADE IV = 20%. The averages of oocytes collected in Group I were (T1 = 7,06 5,10; T2= 7,00 3,64; T3 = 10,60 6,51 and T4 = 2,78 2,59) and the second group (GROUP II) (T1 = 4,90 4,47; T2= 2,93 3,05; T3 = 6,11 4,11 and T4 = 2,70 2,00). For the averages of viable oocytes, the results had been the following ones (T1 = 4,35 3,01; T2 = 4,88 2,96; T3 = 6,87 3,91 and T4 = 1,89 1,90) for GROUP I and (T1 = 2,70 2,22; T2 = 1,92 1,98; T3 = 3,47 2,27 and T4 = 2,00 1,05) for GROUP II. In this experiment, 538 oocytes had been taken to the IVP, 161 (32,85%) of these oocytes had clived and 65 (13,26%) had resulted in Morula or Blastocysts. The statistcs analysis of the results did not demonstrate significant difference in the oocyte number of the diferent groups, caused for the suction pressure. It was observed that the type of needle is related of pressure of the vacuum bomb, the best results were finding with T3 treatment, moreover It can be observed through these findings that the best averages of oocytes collected had been gotten in the group of animals that had presented ovarians cysts (GROUP I), We concluded in this experiment that the use of OPU/IVF technique is an great option in the prolongation of the reproductive life in problems zebu cattle.
Acesso aberto (Open Access)
Avaliação de diferentes diluentes na criopreservação de sêmen ovino (Ovis aries)
(Universidade Federal do Pará, 2010-08-31) GUIMARÃES, Adrianne Araújo; VALE, William Gomes; http://lattes.cnpq.br/7486151987920142
During cryopreservation are numerous changes experienced by the sperm cells, which leads to the decrease in motility and loss of viability after thawing. For this reason, there arises the need to refine the process technology of semen, especially regarding the use of diluents. The aim of this study was to evaluate the viability of spermatozoa undergoing the process of cryopreservation and dilution, using three extenders (TES, TRIS and PBS) and three years of stability (4h, 8h and 12h), and the study was divided into nine groups: TES-4h-8h TES, TES- 12h, TRIS-4h-8h TRIS, TRIS-12h-4h PBS, PBS and PBS-8h-12h. After collection, semen was evaluated macro and microscopically, and pre-diluted solutions in TES, TRIS and PBS, but without the addition of cryoprotectants (Solution A), where they remained for 1 hour. Subsequently, semen was diluted with solutions TES, TRIS and PBS, already containing the cryoprotectants, again assessed to be so packed, and subjected to different periods of equilibrium, and then frozen. After thawing, motility was assessed and vigor, and after the TTR, motility, vigor and detachment of the acrosome. On cryopreservation with TRIS, motility and vigor were statistically similar (p> 0.05) in semen frozen with a balance of 4h (17.2% and 1.6), 8h (22.4% and 2.1) and 12h (14.8% and 1.6) (p> 0.05). After the TTR was not observed statistical difference (p> 0.05) in motility and vigor in 4h (10.4% and 1.1) and 12h (10.0% and 1.2) of balance, but there was an increase in 8h (15.6% and 1.6) (p <0.05). There was no statistical difference (p> 0.05) in the rate of detachment of the acrosome between 4h (35.1) and 12 (37.6) equilibrium, with a reduction in these indices at 8 am (30.4) (p <0 , 2005). On cryopreservation with ERT, the motility of frozen semen with a balance of 4h (24.8%) and 12h (27.6%) were statistically similar (p> 0.05), although the time of 8h (40.4% ) differed significantly from the others. The force was statistically similar (p> 0.05) for thawed-balanced 4h (2.0), 8h (2.6) and 12h (2.3), not statistically different (p> 0.05) . After the TTR was not observed statistical difference (p> 0.05) for sperm motility in 4h (18.8%) and 12h (17.4%) of balance, but there was an increase in 8h (29.6%) (p <0.05). In relation to the force and the detachment of the acrosome was no statistical difference (p> 0.05) 4h (2.0 and 35.8), 8h (2.1 and 33.4) and 12h (1.8 and 42 , 2) equilibrium. On cryopreservation with PBS, showed no motility and vigor after thawing. These results indicate that solvent-based TES and the equilibrium time of 8h were more suitable for dilution and freezing of ram semen.
Acesso aberto (Open Access)
Biometria testicular, caracterização e congelamento de sêmen de macacos-de-cheiro de vida livre (Saimiri vanzolinii, S. cassiquiarensis e S. macrodon) e cativeiro (S. Collinsi) em água de coco em pó (ACP-118®)
(Universidade Federal do Pará, 2014-06-24) OLIVEIRA, Karol Guimarães; DOMINGUES, Sheyla Farhayldes Souza; http://lattes.cnpq.br/2794753357251149
Recently, Saimiri sciureus taxon (squirrel monkey) was revised and some subspecies are now classified by some authors as species, for example: Saimiri collinsi, S. cassiquiarensis and S. macrodon. Phenotypically these species are well determined. However, information on reproductive aspects is still quite scarce. The objective of the first phase of this study was to describe and correlate the testis with the seminal quality from captive (S. collinsi) and free-living (S. vanzolinii, S. cassiquiarensis and S. macrodon) squirrel monkeys. Length, width, height, circumference and testicular volume were measured as well as motility, vigour, plasma membrane integrity and morphology, for correlation data. No difference was observed between the right and left testicles within species, neither difference between species concerning to testicular measures. Semen, which was collected by electroejaculation with rectal probe, was constituted by a coagulated fraction, a liquid fraction or both. Seminal quality was similar among species. There is no correlation between testicular volume and seminal volume (of liquid or coagulated fraction) in S. collinsi and S. vanzolinii. The second phase of this work aimed to propose a semen freezing protocol in an extender based in ACP- 118® (powdered coconut water), testing two glycerol concentrations (1.5 and 3%) for captive S. collinsi. The third phase aimed to test a semen cooling protocol without addition of egg yolk (extracellular cryoprotector) followed by freezing with the best glycerol concentration observed in the preceding experiment and apply these protocols, previously tested in captive S. collinsi, in free-living males from S. vanzolinii, S. cassiquiarensis and S. macrodon in field conditions. Semen from S. collinsi could be cooled in ACP-118® without egg yolk. The freezing protocol developed proved to be useful for semen cryopreservation of S. Collinsi, although requiring improved to allow better maintenance of sperm quality after thawing. The procedure proved to be also applicable under field conditions on free-living animals, providing very encouraging results for the formation of genebanks for species vulnerable to extinction as the S. vanzolinii.
Acesso aberto (Open Access)
Comparação entre o TRIS, Lactose/TRIS, Ringer-Lactato e Leite Desnatado como diluidores na criopreservação do sêmen bubalino
(Universidade Federal do Pará, 2012-02-29) MIYASAKI, Michel Yoshio Almeida; VALE, William Gomes; http://lattes.cnpq.br/7486151987920142
The objective of this experiment was to test the effectiveness of different extenders, the basis of Ringer Lactate, Skim Milk, TRIS (hydroxymethyl-amino-methil Methan) and Lactose/TRIS, the cryopreservation of buffalo semen. We used three male Murrah buffaloes in full sexual activity. The semen was collected by artificial vagina total of 71 ejaculates. After harvest, each sample was subjected to qualitative and quantitative analyzes of semen. The ejaculates were split and diluted in four extenders. The diluted semen was stored in straws of 0,25 mL and subjected to an equilibration time of up to four hours at 5°C, with subsequent freezing in liquid nitrogen. The identified samples were thawed in a water bath at a temperature of 40°C for 30 seconds and subsequently evaluated for motility, vigor, acrosome damage, and percentage of sperm pathologies. The samples were also tested with heat-resistance, while remaining incubated at 40°C for 30 seconds, 3-5 minutes, 30 minutes, 1 hour, 2 hours and 3 hours, which were evaluated for motility and spermatic . The physico-chemical, after fresh semen analysis, were within the prescribed values for buffaloes and satisfactory for the freezing process. After thawing the semen, observed numerical reduction (p<0,05) in sperm motility, with fresh semen was found 86,67±6,17% decreasing to 70±6,92% in TRIS, 67,4±8,01% in Ringer/lactate, 67,09±9,03% in Lactose/TRIS and 59,7±9,05% in skim milk and to compare between the four treatments only the skimmed milk showed a statistically significant difference (p<0,05). After thawing, spermatic vigor also decreased significantly (p<0,05) in four treatments (3,50±0,53 TRIS, 3,38±0,49 Ringer's lactate, 3,3±0,46 Lactose/TRIS and 3,25±0,44 Skim milk) versus (4±0,39 fresh semen) and when comparing between treatment, only between skimmed milk and TRIS was no statistical difference (p<0,05). As for the larger defects (4,15±1,9% fresh semen; 10,51±4,4% TRIS, 11,94±4,2% Ringer's lactate, 11,88±4,8% Lactose/TRIS; 12,01±5% skim milk), smaller (3,81±1,2% fresh semen and 4,67±1,1% TRIS, 4,98±1,7% Ringer's lactate, 4,93±2,0% Lactose/TRIS, 4,93±2,0% skimmed milk) and total (7,91±2,1% fresh semen; 15,18±4,7% TRIS, 16,92±4,8% Ringer's lactate, 16,82±5,6% Lactose/TRIS, 17,11±5,6% skimmed milk), after thawing showed a significant increase of defects in the four treatments (p<0,05), and among them were not statistically different between groups (p>0,05). In the post-TTR after 3 hours of incubation, progressive motility (21,13±7,5% TRIS; Ringer lactate 20,78±7,4%; Lactose/TRIS 20,25±5,3%; Skimmed milk 20,12±6,6%) and spermatic vigor (TRIS 2,04±0,5, Ringer Lactate 2,07±0,5, Lactose/TRIS 2,02±0,4, 2 skim milk, 2,00±0,5) showed no statistical difference between treatments (p<0,05). As for intergridade acrosome after thawing (semen in natura 97,85,±0,6%; TRIS 91,65±4,3%, Ringer-Lactate 90,46±4,8%; Lactose/TRIS 89,76±5,4%; Skimmed milk 90,56±5,6%), decreased (p<0,05) and when comparing this parameter between treatments was not observed statistically significant differences between the treatment (p>0,05). Given the observed results we conclude that the freezing of buffalo semen extenders with TRIS (Tris-hydroxy-methyl-amino-Methan), Ringer Lactate, Lactose / TRIS and skimmed milk showed satisfactory function in cryoprotection of sperm viability semen in different stages of cryopreservation.
Acesso aberto (Open Access)
Criopreservação do sêmen de macaco-prego (Sapajus apella Linnaeus, 1758): avaliação de diferentes diluidores, concentração de glicerol e antioxidante catalase
(Universidade Federal do Pará, 2015-02-24) LEÃO, Danuza Leite; DOMINGUES, Sheyla Farhayldes Souza; http://lattes.cnpq.br/2794753357251149
The main objective of the present study was to establish an efficient semen cryopreservation protocol in Sapajus apella for maintaining sperm viability. For this, we compared the performance of TES-TRIS, CWS and ACP-118® extenders during the seminal coagulum dissolution and cooling. Then, in order to improve the sperm parameters, we also determined the glycerol concentration (3; 5 and 7%) as well as to evaluate the effect of catalase antioxidant (10 µg and 50 µg). Therefore, six adult males of S. apella from the National Primate Center were used. Semen was collected by electro-ejaculation with rectal probe after chemical restraint of animals with ketamine and xylazine hydrochloride. In the first phase of the project, the semen obtained was diluted in TES- TRIS, CWS and ACP®-118 extenders, kept in a water bath at 37 °C. After coagulum dissolution, the semen was cooled in the refrigerator at 4 °C for 90 minutes and evaluated after 28h for motility, vigor and plasma membrane integrity percentage before and after the cooling. ACP®-118 was the best extender to preserve the motility sperm and plasma membrane integrity after 28 hours of incubation. Based on these results, cryopreservation was performed by evaluation of different concentrations of glycerol (3, 5, and 7%). The cryopreserved sperm to 3% glycerol had better results. In the second project phase was performed S. apella semen cryopreservation in ACP®-118 extender supplemented or not with catalase antioxidant (10 µg/mL and 50 µg /mL). All treatments were effective in maintaining sperm parameters and was possible to recovery the post-thaw motility. The catalase 50 treatment was the best for maintenance of the vigor after cooling and plasma membrane integrity in post-thaw sperm. So, we concluded that ACP-118® extender can be efficiently used for S. apella semen cryopreservation added 3% glycerol, moreover, the addition of the catalase antioxidant showed beneficial effect during this process.
Acesso aberto (Open Access)
Efeito do cortisol na produção in vitro de embriões bovinos
(Universidade Federal do Pará, 2014-12-19) ALMEIDA, Nathalia Nogueira da Costa de; MIRANDA, Moysés dos Santos; http://lattes.cnpq.br/3354029928888919; OHASHI, Otávio Mitio; http://lattes.cnpq.br/5547874183666459
The aim of this thesis was to evaluate the effect of cortisol in oocyte maturation and bovine embryo culture. In Chapter 1 we analyzed the distribution and location of glucocorticoid receptor (GR) by immunocytochemistry in oocytes, cumulus cells, embryonic cells 2-4, 8-16 cell, morula and blastocyst, and was also verified the presence of mRNA (qualitative RT-PCR) for GR in the stages. The results showed that the GR is present in all cells analyzed. In order to verify the functionality GR in preimplantation embryo development, translation of mRNA for GR zygotes was silenced in the RNAi technique, and subsequent embryo development was analyzed. The embryonic development decreased (P <0.05) after silencing of GR mRNA. The presence of GR oocyte and the cumulus cells indicates that these cells are sensitive to the use of CG. Given this, in Chapter 2, the effects of different cortisol concentrations during in vitro maturation (IVM) of bovine oocytes and embryonic development, apoptosis rate and gene expression (NRF1, COX, TFAM, GLUT1, FASN and HSP70) were analiyzed. The concentrations of cortisol used were 0.01, 0.1 and 1 mg / mL . There was no statistical difference in the number of cells and cleavage rate, but the concentration of 0.1 mg/ml of cortisol increased the blastocyst rate when compared to the control group (without cortisol in IVM) (41 ± 10 versus 21 ± 1.2; p <0.05, respectively). The rate of apoptosis and gene expression in oocytes, cumulus cells and blastocysts was only assessed at a concentration of 0.1 mg / ml of cortisol. There was no statistical difference in the apoptotic index, and not with respect to gene expression in oocytes and cumulus cells for COX genes, NRF1, HSP70 and FASN (p> 0, 05). Regarding embryonic gene expression, only the measurements relative mRNA FASN, GLUT1 and HSP70 were increased in blastocysts treated with 0.1 g/ml during IVM when compared to embryos of the Control group (p <0.05), the other genes showed no change (p> 0.05). In Chapter 3, we investigated the use of cortisol during in vitro culture (IVC) of bovine embryos. As in Chapter 1 was identified GR in all embryonic stages and when the GR expression was silenced and embryonic development was impaired. Thus, in the experiment three different concentrations (0.01, 0.1 and 1 mg / mL) of cortisol in IVC and embryonic development was evaluated same parameters of Chapter 2. There was no statistical difference in the embryos treated with cortisol in different concentrations when compared to the control for the parameters analyzed (p> 0.05). The concentration of 0.1 mg/ml was chosen to evaluate other parameters of embryo quality. Thus, IVC embryos with or without 0.1 mg/mL of cortisol, were analyzed for apoptosis rate and gene expression, not being statistically significant difference in any of the analyzes (p> 0.05). After these studies conclude that oocytes and embryos are responsive to GC, and the addition of cortisol in IVM improves oocyte competence, but the supplementation of IVC with cortisol may not have influence on embryo development.
Acesso aberto (Open Access)
Expressão gênica e viabilidade de folículos pré-antrais inclusos em fragmentos de córtex ovarianos cultivados in vitro de macaco-prego (Sapajus apella)
(Universidade Federal do Pará, 2011) BRITO, Adriel Behn de; SANTOS, Regiane Rodrigues dos; http://lattes.cnpq.br/0500967766886604; DOMINGUES, Sheyla Farhayldes Souza; http://lattes.cnpq.br/2794753357251149
The aim of the present study was to investigate the stability of three reference genes in the ovarian tissue of capuchin monkeys (Sapajus apella) and to develop a short-term in vitro culture system for the activation and growth of preantral follicles from capuchin monkeys. To this end two experiment were conducted as follow. Experiment I: Fresh and cryoprotectant exposed ovarian biopsies were used. Both fresh and exposed ovarian tissues were subjected to total RNA extraction and synthesis of cDNA. After amplification of cDNA by real-time PCR, the GeNorm, Bestkeeper and Normfinder software were used to evaluate the stability of glyceraldehyde-2-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyltransferase 1 (HPRT1) and TATA-binding protein (TBP). Experiment II: Ovarian tissue from four healthy mature females were collected and divided into nine ovarian cortical pieces of 1 mm³. One ovarian fragment (control) was immediately divided in two pieces, which were subjected to viability analysis or qRT-PCR. The remaining 8 fragments were individually cultured in vitro in a medium consisting of TCM supplemented with 100 ng/mL EGF (T1), either or not added with 10 μM BME (T2), 100 ng/mL BMP4 (T3), 25 IU PMSG (T4), 10 μM BME and 100 ng/mL BMP4 (T5), 10 μM BME, 25 IU PMSG (T6), 100 ng/mL BMP4, 25 IU PMSG (T7) or 10 μM BME, 100 ng/mL BMP4, 25 IU PMSG (T8). Results demonstrated that, in the ovarian tissue from capuchin monkeys, HPRT1 and TBP were the most suitable reference genes and thus could be used as parameters to normalize data in future studies. In contrast, GAPDH appeared as the least stable gene among the tested reference genes. After in vitro culture, all treatments resulted in similar percentages of viable preantral follicles. Ovarian tissue cultured in the presence of EGF + BME/BMP4/PMSG resulted in an increased rate of follicular activation and growth, as well as in the up regulation of AMH, BMP15 and GDF9, specific markers of follicular development. In conclusion, HPRT1 and TBP are the most stable reference genes in fresh and cryoprotectant exposed ovarian tissue from capuchin monkeys. Preantral follicles are able to develop in vitro when cultured in a medium supplemented with PMSG, BME and BMP4. Follicular viability, however, was maintained independently on the culture medium. The use of growth factors as markers of follicular development was crucial to identify the best culture medium.
Acesso aberto (Open Access)
Expressão gênica e viabilidade de folículos pré-antrais submetidos à vitrificação do córtex ovariano de Sapajus apella (macacas-prego)
(Universidade Federal do Pará, 2012-05-25) SANTANA, Luana de Nazaré da Silva; DOMINGUES, Sheyla Farhayldes Souza; http://lattes.cnpq.br/2794753357251149; SANTOS, Regiane Rodrigues dos; http://lattes.cnpq.br/0500967766886604
Vitrification is a biotech that has been increasingly showing promise in several species, among them domestic ruminants (such as sheep, goats and cows) and in nonhuman primates (NHP) (as cynomologus and rhesus), and consists in reducing ultra-rapid temperature by the presence of high concentrations of cryoprotective agents (PCA's) in liquid nitrogen. Thus the aim of this study was to develop a methodology for cryopreservation by vitrification of preantral follicles (PF's) in Sapajus apella. For this purpose, we used females (n = 9) adult Sapajus apella squad belonging to the National Primate Center (CENP). Samples were taken from the ovarian cortex by laparotomy so as not to destabilize the animals reproducibly. The fragments exposed to 8 different treatments: Ethylene glycol (EG) + 40% Sucrose (Sac) dissolved in 0.5 M TCM-199, added to Selenium (2,5, 5 or 10 ng / ml) or Trolox (25, 50 or 100 mM). Following exposure to cryoprotective agents was analyzed follicular viability before and after vitrification, from the follicular morphology, expression of Hsp70 genes, Erp29, Erp60 SOD1 and through the analysis by Polymerase Chain Reaction (PCR), in addition to performing measurement of oxidative stress thru the TEAC (English Total Equivalent Antioxidant Activity). The analyzes showed that vitrification allowed the maintenance of follicular viability by previous exposure to concentrations of ACP's alevadas, especially when supplemented with 50 mM trolox (which resulted in high follicular survival rates) and increased expression of the antioxidant enzyme SOD1. While in the absence of it was observed an increase in rates of follicles degenerate and vacuolated, and reduced expression of the antioxidant enzyme SOD1 and increased expression of chaperone Erp29.
Acesso aberto (Open Access)
Incubação de ovos e criação de filhotes de guará (Eudocimus ruber) no parque Mangal das Garças: uma ferramenta para a conservação da espécie
(Universidade Federal do Pará, 2015-06-12) MIRANDA, Stefânia Araújo; DOMINGUES, Sheyla Farhayldes Souza; http://lattes.cnpq.br/2794753357251149
The population of scarlet ibis is considered to be extinct in several regions. Captive breeding techniques are complementary tools for species conservation. The objectives were to develop a feeding protocol and an ambient environment for the rearing of scarlet ibis chicks, to intensify reproduction in captivity and increase the number of ibises in the colony, describe the developmental stages of chicks and develop growth estimation equations. The chicks were fed with three diets: Diet C (commercial feed), S (commercial ration and shrimp) and F (commercial ration and fish). The body condition and weight were taken daily, and the body measurements every seven days. The mortality rate under Diet C was 100%, and 0% under Diets S and F. The mean body weight and bones lengths were higher with Diet F and the mean length of the unpigmented portion of the beak did not differ among the diets (P<0.05). Diet F had higher nutritional contents. Rearing was artificial during year I and natural during year II at one of the areas of the park. The limbs development and body condition were evaluated daily, and the weight every seven days during artificial rearing. The mortality rate was lower and the number of births and survival rate were higher during artificial rearing. All of the young exhibited weight gain (P<0.05), good body condition and no dystrophies were observed. The development stages of chicks and the weight were monitored daily, and the measurements of the beak and bones were taken every seven days. The eyes opened on day 4.73±0.12, the nestlings moved within the nest on day 6.31±0.18 and left the nest on day 15.3±0.68. The means weight and measurements of the bones increased (P<0.05). The means of beak despigmentation decreased (P<0.05). The beak pigmentation started on day 7º and on day 35º was completely pigmented. The correlation between all parameters and the determination coefficients of regression equations were high. Diet F was found to be best for the artificial rearing. The artificial rearing protocol provided an adequate environment and feeding, intensified the reproduction and increased the number of scarlet ibises. Growth estimation equations were developed to assess the chicks growth and was possible to describe the developmental stages. Beak pigmentation was found to be a useful parameter for estimating the age.
Acesso aberto (Open Access)
Polimorfismos na região promotora, Éxon I e Éxon II do gene do receptor da melatonina associados às características reprodutivas em búfalas na Amazônia
(Universidade Federal do Pará, 2015-06-18) BARBOSA, Elizabeth Machado; RIBEIRO, Haroldo Francisco Lobato; http://lattes.cnpq.br/1614582293203770
The buffaloes production in the Pará state have is a high representative activity to the regional economy and production of meat, milk and dairy products. Therefore, it is necessary to develop new technologies that increase the enjoyment of the production and reproduction of buffaloes in the Amazon region. Thus, the objectives of this research thesis was to characterize the promoter region of the melatonin receptor gene 1 (MTRN1A) and identify polymorphisms in exons I and II of that gene and link them to reproductive traits that have economic importance in buffaloes in the region Amazon. Were collected 400 buffaloes samples and 60 animals were used to characterize the promoter region, 140 to detect the polymorphism (SNPs) in exon II and 77 to detect the polymorphism in exon I. DNA extraction was peformed by phenol method. Then were selected different primers to make Polymerase Chain Reaction (PCR). The promoter region was sequenced MTRN1A a total of 1621 base pairs by the Sanger method, polymorphism in exon II was detected by PCR-RFLP (Restriction Fragment Length Polymorphism) with HpaI enzyme and the polymorphism exon I was found by sequencing DNA by Sanger method. Allele and genotype frequencies of all SNPs were evaluated, the inbreeding coefficient (Fis) and the likelihood of Hardy-Weinberg equilibrium. SNPs in the promoter region and exon I have been associated with reproductive traits of buffaloes by ANOVA or Student's t test and / or chisquare test. The significance level was 0,05. 26 SNPs were detected in the promoter region at positions -1511 (C → T), -1465 (G → T), -1422 (A → G), -1411 (G → A), -1395 (G → T), - 1298 (A → G) -1295 (G → A) -1242 (C → A) -1150 (C → T), -1 147 (G → C) -1136 (A → G) -911 ( G → A), -909 (A → G) -724 (C → G) -656 (A → C) -649 (C → T), -644 (G → A), -511 (A → C) -481 (G → A), -425 (C → T) -395 (G → A), -383 (G → T), -254 (C → T) -206 (T → C) , -133 (T → G) and -94 (C → T) and a deletion unit (ACAA) at position -1483. Of the total detected SNPs, 75% of the wild alleles tiveral frequencies greater than 0,5. For the characteristic interval between calving (IEP), only five SNPs (-1298, -1136, -911, -724 and - 656) were significantly associated (P <0.05) and three SNPs (-1395, -724 and -94) were significantly associated (P <0.05) with the characteristic age at first calving (IPP) and none for the characteristic concentration of deliveries (P> 0.05). A total of 11 SNPs was strongly associated with binding factors in gene regulation. The SNP in exon II by PCR-RFLP (HpaI) at position 306 (T → C), the most frequent allele T in Upland animals (0,529) and C in lowland animals, the two populations showed coefficients Fis inbreeding (0,040 and 0,091, respectively) and strong deviations from Hardy-Weinberg equilibrium (P> 0.05). The mutation occurs at codon 106 and is the sinonímio type without changing the messenger RNA handles structure. The SNP in exon I was detected at position 62 (T → C) and of the non sinonímio, exchanging of Leucine to Proline. The mutant allele was the most frequent C (0,623) inbreeding coefficient Fis = 0.397 and Hardy-Weinberg deviations (P <0.05). None of the genotypes were associated with calving interval and age at first birth (P> 0.05). Therefore, SNPs are strong candidates for selection, but it would be interesting to evaluate them in other herds in the Amazon region and / or in other regions of Brazil and / or other countries to effect them as excellent molecular markers for reproductive traits of buffalo.
Acesso aberto (Open Access)
Polimorfismos no gene da osteopontina e suas associações com a fertilidade de búfalos na Amazônia
(Universidade Federal do Pará, 2012-09-24) ROLIM FILHO, Sebastião Tavares; RIBEIRO, Haroldo Francisco Lobato; http://lattes.cnpq.br/1614582293203770
Because the osteopontin gene can influence the fertility of water buffaloes, the aim of this work was to identify polymorphisms in this gene and associate them with fertility parameters of animals kept under extensive grazing. We used 306 male buffaloes older than 18 months, bred on two farms, one in the state of Amapá and the other in the state of Pará. We identified three SNP for the region amplified by the primer OS4 (5`UTR) and four SNP polymorphisms for the region amplified by the primer OS9 (exon 5 to exon 6). The polymorphisms were in positions 1478, 1513 and 1611 in the region amplified by OS4 and positions 6690, 6737, 6925 and 6952 in the region amplified by OS9. We calculated correlations with the traits scrotal circumference and volume and sperm motility, concentration and pathology. There were SNPs for all the traits studied at 5% significance: for circumference, SNP 6690; for volume, SNP 6737; for concentration, SNP 6690; for motility, SNP 6690; and for pathology, SNP 6690. Therefore, SNP 6690 was related to four traits. The AA genotype of SNP 6690 presented the highest averages for scrotal circumference, sperm concentration and motility and the lowest total number of pathologies. However, for the scrotal volume trait, the animals with the largest volume were correlated with the presence of the GG genotype of SNP 6737. This indicates that the osteopontin gene is important because it can have a substantial influence on the reproductive traits of male buffaloes.
Acesso aberto (Open Access)
Puberdade em caititus (Tayassu tajacu): estudo da espermatogênese em diferentes faixas etárias
(Universidade Federal do Pará, 2009-05-20) CARDOSO, Deise de Lima; FERREIRA, Maria Auxiliadora Pantoja; http://lattes.cnpq.br/1832728101486131; GUIMARÃES, Diva Anelie de Araújo; http://lattes.cnpq.br/2891287458034896
This study analyzes the development of spermatogenesis in peccaries (Tayassu tajacu) and establishes the age the reach puberty, considering testicular biometry and seminiferous tubules, the amount of spermatogenic cells, the morphological description of the stages of the seminiferous epithelium cycle (SEC), the relative frequency on which they surge inside the seminiferous tubules and general spermatogenesis yield. In the experiment, the animals were divided according to age bracket, into five groups, with three animals in each group, G1 (7 to 8 months), G2 (9 to 10 months), G3 (11 to 12 months), G4 (13 to 14 months) and G5 (15 to 16 months). The animals were subjected to orchiectomy surgery, to obtain the testicle samples, which were fixed in Alfac solution for 24 hours and histologically processed, where 5 μm cuts were stained in hematoxylin eosin. Based on the tubular morphology method, it was done the quantification of the cell types corrected to its average nucleus diameters of 10 transversal sections at SEC stage 1 for animals with established spermatogenesis and 20 transversal sections for the younger animals without any established spermatogenic activity; and also the classification of the stages of the seminiferous epithelium cycle, by analyzing 100 transversal sections of seminiferous tubules. The data from the testicular biometry, such as, weight, length and width, showed gradual and constant growth with significant statistical differences (p<0,05) and high correlation among themselves. The tubular diameter values presented statistical significance from G1 to G4, and from G4 an accelerated and continuous growth, with no statistical significance (p>0,05). According to the quantitative and morphological analysis of the spermatogenic cells which form the germinative epithelium, the animal age brackets were classified in the following phases impuberal (G1), prepuberal (G2), puberty (G3), pos puberty 1(G4), and pos puberty 2 (G5). The phase the productive activity started, or, puberty, was determined in the animals when they turned 11 months, when the the greatest growth in the number of spermatogenic cells and a positive correlation with testicular weight occurred. At this stage, the Sertoli cells, presented a significant decrease (p<0,05). During determination of relative frequency of the stages of seminiferous epithelium cycle, eight types of association were observed, according to the tubular morphology method, where the stages with greater and less frequency were 1 and 3 respectively. The pos meiotic phase showed more frequency and the meiotic phase, the lower one, being statistically significant in relation to the other ones. The reproductive efficiency was demonstrated through translated values by the cell ratios between type A spermatogonic cells and round spermatids, being that no significant increase between G3, G4 and G5 was observed (p>0,05); and the rate of Sertoli cells showed a significant statistical difference between all possible comparisons of age brackets from G3 to G5 (p< 0,05).
Acesso aberto (Open Access)
Respostas termográficas em touros bubalinos submetidos à coleta de sêmen e avaliados sob condições agrometeorológicas no trópico úmido
(Universidade Federal do Pará, 2014-04-30) BARROS, Daniel Vale; LOURENÇO JÚNIOR, José de Brito; http://lattes.cnpq.br/2919433679918544; GARCIA, Alexandre Rossetto; http://lattes.cnpq.br/2678267039338224
Buffalo (Bubalus bubalis) livestock is mostly performed in the intertropical zone, where high temperatures prevail. Therefore the knowledge about buffaloes physiology on tropical environments and their possible responses due to tropical climate changes are essential. The objective of the study was to evaluate the variation in thermal comfort, physiological, hematological, seminal parameters and the superficial temperatures of buffalo bulls raised on humid tropical climate (Afi, Köppen classification). Ten buffaloes were kept in collective paddocks with free access to shade. During five months, data were registered from climatological meteorological station and three distinct dataloggers installed inside the stalls for calculating the Temperature and Humidity Index (THI). Respiratory rate (RR), heart rate (HR), rectal temperature (RT), superficial temperature of the eye (GLO), superficial temperature of the scrotum (ESC), superficial temperature of the right flank (FLd) and left flank (FLe) were registered. The Benezra´s Comfort Index (ICB) was also calculated. Semen collection was performed weekly by artificial vagina and blood sampling for assessment of blood counts were done monthly. The mean maximum of air temperature was 31.5°C and maximum a verage relative humidity was 93.2%. The THI was different only between periods (P<0.05). The RR, HR and ICB showed significant difference over the months and between shifts (P <0.05). RT differed between periods and reduced along months with lower value in August (37.8 ± 0.7°C). RT, GLO, FLd, FLe and ESC showed no diffe rence (P<0.05) for both periods and months. The hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, gross motility and sperm vigor showed significant differences (P<0.05) along the months. The highest correlations between THI and superficial temperatures were between ITUmed and FLdmed (0.77; P<0.0001), ITUmed and FLemed (0.75; P<0.0001), ITUmed and GLO (0.72; P<0.0001), and ITUmed and ESC (0.41; P<0.0001). The highest correlation between internal temperature and surface temperature was TR and GLOmax (0.58; P<0.0001). Significant correlations were found between ICB and FR (0.97; P<0.0001), ICB and FC (0.89; P<0.0001), FC and FR (0.87; P<0.0001), THI and integrity of the plasma sperm membrane (-0.17; P<0.05). The results showed that animals had variations in thermal comfort and increased superficial temperature in the hottest periods of the day, however they were able to maintain homeothermy. Finally, the infrared thermography can be used as a noninvasive and auxiliary technique in studies about animal physiology and thermoregulation.
Acesso aberto (Open Access)
Respostas termolíticas e qualidade seminal de ovinos naturalizados criados em ambiente tropical
(Universidade Federal do Pará, 2015-10-30) KAHWAGE, Priscila Reis; GARCIA, Alexandre Rossetto; http://lattes.cnpq.br/2678267039338224
The study aimed to deepen knowledge about anatomical and physiological features related to body heat loss ability and relation to semen quality of naturalized sheep Morada Nova and Santa Ines on tropical environment. Nine male Santa Inês and seven Morada Nova breed were subjected to two experiments. The first test included monitoring of coat features (amount of melanin, thick, length and diameter of the fibers), sweat rate, physiological indicators of thermal comfort (respiratory rate, heart rate, rectal temperature, surface temperature, skin temperature and scrotal surface temperature), sperm quality (sperm concentration, progressive motility, plasma membrane integrity, major defects, minor defects and total defects) and testicular parenchyma integrity coupled with the meteorological variables measured throughout the year. Statistical analysis was performed using the GLM and LSMEANS procedures of SAS software, version 9.1.3. Significant differences (P <0.05) were observed between the summer and fall and winter, with increase in termolitics parameters during the hot season. Already sperm quality did not vary throughout the year. The second experiment evaluated the capacity of rams to maintain body and testicular homeothermy under thermal challenge. Animals were subjected to heat tolerance test, which consists maintenance in the shade (period 1), followed by exposure to the sun (period 2) and return to the shadow (period 3). In the three periods were measured: respiratory rate, heart rate, body surface temperature, back, eyeball and scrotum, by infrared thermography. Adaptability of the animals was measured by heat tolerance index calculation. Sperm quality and testicular parenchyma integrity were evaluated before and after the heat challenge. In Period 1, the variables had baseline values for both genotypes. In Period 2, the variables involved in thermolysis significantly increased (P<0.05), which matches a thermal discomfort situation. In Period 3, the variables returned to baseline values and some values were lower than those in Period 1. Seminal and ultrasonographic variables did not share with thermal insult. It is concluded that MN and SIN rams had efficient thermolytic mechanisms that favor preserving gonadal functionality. The animals were considered resilient to a thermal challenge.
Acesso aberto (Open Access)
Ultrassonografia testicular, em machos bubalinos criados em regime extensivo no estado do Pará
(Universidade Federal do Pará, 2011-07-07) MANRIQUE AYALA, Henry Daniel; RIBEIRO, Haroldo Francisco Lobato; http://lattes.cnpq.br/1614582293203770
The objective was to verify the biometrics associated with testicular ultrasound as a tool in the selection of sexual precocity in male buffalo. We used 19 male crossbred Murrah and Mediterranean breeds, aged between 11 and 59 months, from May 6 to November 18, 2010. The animals were divided into 12 age groups: 8 animals from 12 to 19 months, 3 animals from 20 to 29 months, and 8 animals from 30 to 59 months. The animals were kept under extensive system in the low land to pasture canarana (Eriochloa SP) without mineralization. The animals were subjected to clinical andrological examination with semen collection by transrectal massage of the ampoules and physical analysis of the ejaculate. The sonographic examinations were performed with the device model Mindray DP-2200, 75L50EAV transrectal linear transducer, multi-frequency 5.0 / 7.5 / 10.0 MHz echodensity The testicular (Ecot) was expressed in number of pixels / area, using the program Image J. We collected blood samples to measure testosterone levels. Statistical analysis was performed using the SAS statistical software 1999. The results of scrotal circumference show averages and standard deviations minimum and maximum of 12.88 ± 0.51cm for animals of 12 and 13 months and 28.38 ± 0.38cm in males aged greater than or equal to 60 months, respectively. The testicular volume measurements showed minimum and maximum values of 30.28 ± 17.37cm³ in animals of 12 and 13 months and 534.25 ± 25.36 cm³ in animals greater than or equal to 60 months, respectively. The mean and standard deviation of the sperm concentration maximum and minimum found were 5.5 ± 3.5 (x 10⁶ / mm³) for animals of 16 and 17 months and 51.41 ± 2.26 (x 10⁶ / mm³) for greater than or equal to 60 months , respectively. Sperm motility in the minimum and maximum percentages were obtained 10% for animals with 16 and 17 months and 80 ± 5.77% to greater than or equal to 60 months, respectively. The total sperm defects found were minimum and maximum of 18.5% in animals of 16 and 17 months and 3%. in greater than or equal to 60 months, respectively. Testosterone levels were found minimum and maximum of 0,070 ± 0,026 ng / ml for 12 and 13 months and 2762 ± 0457 ng / ml for animals greater than or equal to 60 months, respectively. The echogenicity for animals aged 12 to 13 months was 78.67 ± 6.36 pixels, in the range of 14 and 15 months was 94.22 ± 3.40 pixels, for animals aged 16 and 17 months was 88.16 ± 3.95 pixels, animals between 18 and 19 months was 96.09 ± 3.40 pixels, for animals of 20 and 21 months was 103.12 ± 3.86 pixels, for 22 and 23 months was 98.4 ± 5.87 pixels, for the age of 24 to 29 months was 114.05 ± 2.42 of pixels, aged 30 to 35 months was 109.24 ± 3.13 pixel for animals of 36 at 41 months was 98.67 ± 5.3 pixel, between 42 to 47 months the average was 99.33 ± 2.1 pixel, between 48 to 59 months was 96.17 ± 1.90 pixel and animals greater than or equal to 60 months of age was 90.13 ± 1.77 pixel. We conclude that ultrasonography, the interpretation of the echogenicity, associated with the clinical andrological examination, is a tool that can be used in the selection of precocity and fertility evaluation of male buffalo.
Acesso aberto (Open Access)
Utilização de células de trofoblasto de embriões partenogenéticos na descrição de haplótipos de BoLA-DRB3-DQA-DQB
(Universidade Federal do Pará, 2015-03-30) SÁ, André Luiz Alves de; MIRANDA, Moysés dos Santos; http://lattes.cnpq.br/3354029928888919
The prospect of increasing the productivity of cattle by means of genetic improvement is increasingly investigated. Among the most studied and and promising genomic regions is the Major Histocompatibility Complex (MHC), since it includes key genes of the immune response of the animals, being able to select makers in this region for increased disease resistance and therefore greater production. The diversity of the cattle MHC, known as BoLA (Bovine Leukocyte Antigens), has been primarily studied using variants of serological specificities or sequencing some of its genes. Most cattle are heterozygous at BoLA and only in special occasions (homozygous animals or animals for which pedigree information is available) can the haplotypes be characterized. Therefore, this study aims to develop a novel approach to describe BoLA haplotypes from heterozygous cows, using trophoblast cells from parthenogenetic bovine embryos derived from slaughterhouse ovaries. Two methods for validating this approach were used: a panel of 445 SNPs spanning BoLA region was informative on the effect of meiotic recombination on the region zigosity; and the comparison of BoLA-DRB3 alleles between the dam and its parthenogenetic embryo derived trophoblast cells (PEDTC) proved to be a reliable and practical method for investigating BoLA homozigosity. Using both methods, the approach presented here was validated, since BoLA homozygous PEDTC were derived from heterozygous cows, allowing the description of BoLA haplotypes. Detailed analysis of the BoLA Class IIa region identified 18 different BoLA-DRB3-DQA-DQB haplotypes, including 16 novel haplotypes. Furthermore, two DQA and one DQB alleles included in these haplotypes were novel. This method was more efficient than to look for homozygous cows or infer haplotype composition based on pedigree information, in addition to avoid ambiguities on the results. New researches aiming for the improvement of this method can increase its efficiency and make it more easily applicable for a variety of genetic studies, using different species and for other purposes.
Acesso aberto (Open Access)
Viabilidade da dose fracionada de sêmen bovino criopreservado descongelada por diferentes métodos
(Universidade Federal do Pará, 2010-05-21) ARAUJO, Gilson Ferreira de; RIBEIRO, Haroldo Francisco Lobato; http://lattes.cnpq.br/1614582293203770
Given the low sperm concentration necessary to promote the fertilization of oocytes in FIV, this paper aims at evaluating the quality of dose of bovine semen cryopreserved after its splitting into two parts to make better use of it on the PIV program. The experiment used 110 doses of frozen semen in pallets of 0.25 mL from 10 bulls, which were divided into stages: A dose represented the control group, five were assigned to the group of direct thawing (DT) and 5 for the group of indirect thawing (IT). Doses were fractionated into two parts, forming two subgroups for each group, the unit sphere (UE) and polyvinyl alcohol unit (PAU). The first subgroup analysis was immediate, and the second was analyzed after 24h storage in N2. Results were obtained through descriptive statistics, Kruskal-Wallis and SNK (P <0.05), motility and force: immediate Indirect thawing: 48.8% / 2.4, IT-24: 54.2% / 2.5, immediate direct thawing: 67.8% / 3.0, direct thawing-24: 69.6% / 3.0, Control: 70% / 3.0. For acrosome injury and membrane integrity: immediate Indirect thawing: 24.3% / 40.5%, indirect thawing- 24: 36.2% / 44.3%, immediate direct thawing: 12.8% / 63.6%, DT -24h: 12.6% / 59.0%, control: 11.9% / 57.5%. Average sperm concentration: DI-immediate: 6.1 × 106 DI-24: 7.2 × 106, DD-immediate: 8.2 × 106, DD-24: 7.8 × 106 sperm / dose fraction. Thus for the fractionation of the dose of bovine semen cryopreserved best thawing method was straightforward, since it showed the best results in all parameters.

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