Livestock genome maps are used to identify economic trait loci and loci proximal to genes affecting economically important traits. This research contributes to the development of the ovine genome map by establishing techniques to physically map large DNA inserts to ovine chromosomes using fluorescent in situ hybridization. Fluorescent in situ hybridization techniques were established using yeast artificial chromosomes as DNA probes. Probes were as large as 1,000,000 kilobases and hybridized to fixed ovine metaphase chromosomes from a callipyge ewe. Three yeast artificial chromosome probes contained a genetic marker for the ovine callipyge gene. Probes were assigned to ovine chromosome pair 18, which was distinguished by a fluorescent R-band pattern. This method verified the location of the marker, which had previously been assigned to chromosome 18 through linkage analysis. This study also determined whether a chromosomal alteration is the cause of the callipyge phenotype, an economically advantageous trait where muscle mass is increased and fat deposition is decreased. A fibroblast cell line was produced from a six-month-old callipyge ewe. Chromosomes were examined using G and R dynamic banding techniques. The ewe had a normal diploid chromosome number of 54, and the karyotype consisted of 3 pairs of submetacentric chromosomes, 23 pairs of acrocentric autosomes, and 2 large acrocentric X chromosomes. G and R banding did not reveal any chromosomal structural abnormalities at the level of resolution (premetaphase) used in this study.

A series of trials was carried out to examine the role of prolactin (PRL) in the control of lactogenesis in New Zealand Romney x Border Leicester ewes. In addition, a study was made of differences in milk yields and plasma PRL concentrations between spring- and autumn-lambing ewes.Daily subcutaneous injections of 2 mg CB154 inhibited PRL secretion and delayed lactogenesis. There were no consistent effects on plasma progesterone or insulin concentrations. CB154 treatment was more effective in reducing milk yield in twin-bearing than in single-bearing ewes when used for 20 days than for 9 days prepartum. The differential effects on milk yield cannot be explained by corresponding effects on plasma PRL or insulin concentrations. Circulating PRL during the period 20 to 10 days prepartum may have an important effect on milk yield in twin- but not single-bearing ewes.Subcutaneous injections of 0.5 mg/kg live weight oPRL, administered on 2 consecutive days peripartum, to ewes treated with CB154 for 7 days prepartum, resulted in milk yields similar to those in control ewes and significantly (P<0.01) greater than those in ewes treated with CB154 alone. This indicated that oPRL prevented the CB154-induced reduction of milk yields and has established that the effect of CB154 on lactogenesis is mediated through suppression of PRL secretion and not by effects on some other hormone.Injection of 10 mg oPRL directly into one mammary gland (via the teat duct) increased milk yields relative to the contralateral, bicarbonate-treated gland in CB154-treated ewes. The intramammary oPRL injection did not raise circulating PRL concentrations. Furthermore, the milk yields of bicarbonate-treated glands in ewes treated with bicarbonate only, did not differ from those of bicarbonate-treated glands in ewes treated with oPRL in the contralateral gland, demonstrating that there were no effects of oPRL, transferred via the circulation from the treated gland, on the contralateral gland. Glands treated with oPRL produced 15% (P<0.05) more milk than the bicarbonate-treated glands during the first 8 days of lactation and the difference was maintained throughout the 8-week lactation period, indicating that the oPRL had effected a permanent change in the ability of the gland to produce milk. It is concluded that PRL acts directly on the mammary gland without the need for a putative intermediate hormone, and that intramammary PRL concentrations during lactogenesis may have long-lasting effects on lactation.The possibility was examined that dietary differences were responsible for seasonal differences in plasma PRL concentrations, milk yields, milk composition, lamb birthweight and lamb growth rate, observed in earlier trials. Mean plasma PRL levels were significantly (P<0.01) higher in spring- (192±38 ng/ml) than in autumn- (71±17 ng/ml) lambing ewes housed indoors under constant photoperiod (18L:6D) and fed the same diet. Milk yields were also significantly (P<0.05) higher in the spring- (2041±114 g/d) than in the autumn- (1563±109 g/d) lambing ewes over the 8 day lactation. Lamb growth rates (adjusted for birthweight, birthrank and sex of lamb) from birth to 8 weeks of age were significantly (P<0.001) higher in spring (282±12 g/d) than in autumn (225±15 g/d). The seasonal differences were confounded with corresponding differences in ewe live weight and it was not possible to determine whether dietary differences contributed significantly to the differences observed.Two routes of oPRL supplementation were used to test the effectiveness of elevating peripheral or local levels of PRL in autumn-lambing ewes which, based on previous results, were expected to have low plasma PRL concentrations and milk yields relative to spring-lambing ewes. Administration of 10 mg supplementary oPRL directly into the gland or subcutaneous injection of 0.5 mg/kg oPRL did not increase the milk yields, or change the composition of milk, compared to controls. These results suggest that the circulating level of PRL, and the intramammary concentration of PRL, in autumn-lambing ewes are not limiting lactogenesis. Because the plasma prolactin concentration in the ewes was unexpectedly high, it was not possible to reach firm conclusions regarding possible effects of supplementary oPRL in ewes with naturally low plasma PRL concentrations. Nevertheless, the results indicate that raising the intramammary concentration of PRL around the time of parturition, in ewes with circulating PRL levels characteristic of normal spring-lambing ewes, does not enhance lactogenesis.It is concluded that PRL is important to the complete initiation of lactogenesis in ewes, that it acts directly on the gland and that it is necessary for establishing the maximum potential of the gland to secrete milk.

La mélatonine est une hormone synthétisée et sécrétée uniquement la nuit par la glande pinéale. Son rôle principal est son implication dans la synchronisation de la saison de reproduction. La mélatonine se lie aux récepteurs, MT1 et MT,, membres de la famille des récepteurs à sept domaines transmembranaires couplés aux protéines G (RCPG).Le clonage récent du récepteur ovin MT2a remis en cause toutes les données connues. La pharmacologie et les voies de signalisation du récepteur oMT2ont été étudiées et sont communes à celles des récepteurs des autres espèces. En revanche, oMT2possède une originalité de séquence avec la présence du motif DRY, fortement impliqué dans l’établissement de l’activité constitutive des RCPG. D’une part, nous avons montré que l’ensemble des récepteurs MT possèdent une activité constitutive. D’autre part, nous avons identifié deux agonistes inverses pour les récepteurs hMT2, initialement décrits comme antagonistes. Dans l’optique de discriminer les rôles respectifs des récepteurs MT in vivo, le développement d’une approche par ARN interférents a été validée dans un modèle cellulaire, la lignée CHO-KI exprimant les récepteurs ovins et de rat. / Melatonin is a hormone synthesized and secreted only during night by pineal gland. A main role of melatoninconcerns its implication in the synchronization of reproductive seasonality. Binding sites of melatonin are MT1and MT2 receptors which belong to the superfamily of seven-transmembrane-spanning G protein-coupledreceptors (GPCRs).Recent cloning of ovine MT2 receptor has challenged the knowledge about melatonin receptors. Wedemonstrated that its pharmacology and signalling pathways were similar to subtype 2 receptor of othersspecies (human and rat). Nevertheless, oMT2 receptor possesses a particularity of sequence, with the presenceof DRY motif which is known to be involved in the establishment of constitutive activity of GPCRs. In ourstudy, we demonstrated the existence of constitutive activity for ail the melatonin receptors. In addition, weidentified two inverse agonists for human MT2 receptors, previously described as antagonists. To describe therespective roles of each subtype of melatonin receptors in vivo, siRNA approach was developed in cell line,CI-10-K Iexpressing ovine and rat melatonin receptors

Made available in DSpace on 2014-06-11T19:27:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-27Bitstream added on 2014-06-13T20:36:34Z : No. of bitstreams: 1 costa_jpr_me_jabo.pdf: 297970 bytes, checksum: 67a31437d11a8fe0ae60126e44cbc31e (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Avaliou-se o efeito da intensidade de pastejo com e sem suplementação no perfil metabólico e indicadores de estresse em 64 cordeiros mestiços Santa Inês (15±3,31). O experimento foi avaliado em medidas repetidas no tempo em um delineamento inteiramente casualizado em esquema fatorial 4 x 2, sendo quatro intensidades de pastejo (IP) (0,8; 1,4; 2,0 e 2,6 de índice de área foliar residual) e grupos suplementados (SP) ou não (NSP). Foram realizadas coletas de sangue a cada 21 dias totalizando seis durante o período experimental. Foram mensurados a hemoglobina, creatinina, uréia, proteína total, albumina, globulina, glicose, ácidos graxos não esterificados (AGNE), beta-hidroxibutirato, colesterol, cálcio (Ca), fósforo (P), magnésio (Mg), relação neutrófilos:linfócitos (N:L) e cortisol plasmático. Nenhum dos indicadores sanguíneos do metabolismo protéico foi afetado pela IP e SP (p>0,05), exceto a albumina que foi maior (p<0,05) nos grupos que receberam suplemento. Os níveis da glicose foi maior (p<0,05) nos grupos suplementados e aumentou com a diminuição da IP (p<0,05). O AGNE apresentou níveis elevados nos grupos NSP (p<0,05) e reduziram com a diminuição da IP (p<0,05). O beta-hidroxibutirato foi modificado apenas pelo SP que foi maior (p<0,05) nos grupos que receberam suplemento. Os teores de Ca apresentaram-se crescentes com o decréscimo da IP (p<0,05). A concentração de P foi maior nos grupos SP (p<0,05). A N:L foi maior nos grupos NSP (p<0,05) e aumentou com o acréscimo da IP (p<0,05). O nível plasmático de cortisol foi maior nos grupos NSP (p<0,05). Esta caracterização metabólica mostrou que a IP e SP modificou o perfil metabólico e os indicadores sanguíneos de estresse de cordeiros em terminação / Evaluated the effect of grazing intensity with supplementation or nonsupplementation in the metabolic profile and stress indicator in sixty four crossbred Santa Inês lambs (15,40±2,31 kg). The trial was evaluated in repeated measurement in a completely randomized design arranged 4 x 2 factorial in which grazing intensity (GI) (0.8 vs. 1.4 vs. 2.0 vs. 2.6 residual leaf area index), supplementation (SP) (nonsupplementation vs. supplementation) were the main effects to be investigated. Six sample of blood were taken during, every 21-days from the jugular vein. The hemoglobin, creatinine, urea, total protein, albumin, globulins, glucose, nonesterified fatty acids (NEFA), β-hydroxybutyrate, cholesterol, calcium (Ca), phosphorus (P), magnesium (Mg), neutrophil: lymphocyte rate (N:L) and cortisol were all measured. No indicator protein metabolism was not affected for GI and SP (p>0,05), except albumin, which was high (p<0,05) in supplemented groups. The glucose levels was high in supplemented groups (p<0,05) and increased with a decrease of GI (p<0,05). The NEFA showed high levels in non supplemented groups (p<0,05) and increased with a decrease of GI (p<0,05). The β-hydroxybutyrate levels was modified only for SP (p<0,05) and supplemented groups showed greater concentration. The Ca levels showed increase with decrease of GI (p<0,05) in both supplemented and non supplemented groups. The phosphorus levels was high in supplemented groups (p<0,05). The N:L rate was modified for GI (p<0,05) showed increased levels with the increase of GI and presented high in non supplemented groups (p<0,05). The levels of cortisol was modified for supplementation only (p<0,05). This metabolic characterization showed clearly the grazing intensity and supplementation modified the metabolic profile and stress indicator in finishing lambs

Made available in DSpace on 2014-06-11T19:23:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-19Bitstream added on 2014-06-13T19:09:59Z : No. of bitstreams: 1 silva_aa_me_botfmvz.pdf: 329313 bytes, checksum: acaef38ccfb3815a5cbf93f92e3438da (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Avaliou-se a segurança e a eficácia da técnica de biopsia pulmonar transtorácica percutânea e seus efeitos sobre o metabolismo oxidativo na espécie ovina. Foram utilizados 20 ovinos clinicamente sadios e destinados ao abate. Inicialmente, foram coletadas amostras de sangue da veia jugular em tubos de ensaio contendo heparina. Após a colheita, foi realizado o exame clínico seguido da biopsia pulmonar com agulha semi-automática no 7º espaço intercostal direito cerca de 5 cm acima do olécrano. Avaliou-se o número de tentativas para a realização do procedimento e o tamanho dos fragmentos. As amostras obtidas foram analisadas histologicamente. Uma nova colheita de sangue foi realizada 30 minutos após a biopsia, e as amostras levadas ao laboratório para a detecção indireta dos radicais livres [superóxido dismutase (SOD), glutationa total (GSH-t) e peroxidase (GSH-Px) e substâncias reativas ao ácido tiobarbitúrico (TBARS)]. Após o abate foi realizado o exame pos mortem para avaliação de complicações da técnica e das eventuais lesões provocadas. Entre todos os animais submetidos à biopsia apenas dois demonstraram resistência à técnica, sendo obtidos fragmentos pulmonares que mediram entre 4 mm a 7 mm de comprimento, com média de 1,8 1 tentativas por fragmento. As principais alterações relacionadas à técnica foram: tosse, dispnéia inspiratória, elevação das freqüências cardíaca e respiratória e aumento do ruído broncobronquiolar. Ao exame pos mortem observou-se hemorragia dos músculos intercostais e pleura visceral de tamanho variado em todos os cordeiros, além de feridas lineares de até 14 mm nos dois animais que não reagiram bem ao procedimento. Das 20 tentativas de execução da técnica, 18 obtiveram sucesso, enquanto que em duas o fígado foi equivocadamente puncionado. As amostras de tecido pulmonar foram consideradas representativas... / It was evaluated the safety and effectiveness of the percutaneous transthoracic lung biopsy technique and its effects of the procedure on oxidative metabolism in sheep. It was used 20 clinically healthy sheep. Initially, blood samples were collected from the jugular vein in test tubes containing heparin. After collecting the blood, the clinical examination was carried out followed by lung biopsy with a semi-automatic needle biopsy. The number of attempts and size of the fragments were measured and the samples were analyzed histologically. New blood samples were collected 30 minutes after the biopsy. All blood samples were taken to the laboratory for the indirect detection of free radicals [superoxide dismutase (SOD), total glutathione (GSH-t), glutathione peroxidase (GSH-Px) and thiobarbituric acid reactive substances (TBARS)]. After the slaughter, a post mortem examination assessed the technical complications and possible injuries. Among all animals underwent biopsy only two did not tolerate the technique. It were obtained Lung fragments from 4 mm to 7 mm and it were required on average 1,8 attempts. The main changes related to the procedure were: cough, labored breathing, increased heart and breathing rate and increased broncobronquiolar noise. The post-mortem examination revealed bleeding of the intercostal muscles and visceral pleura of various sizes in all lambs, and a wound up to 14 mm in the lung of two animals that did not tolerate the procedure. From 20 attempts to implement the technique, 18 were successful, while in two of them the liver was mistakenly punctured. Samples of lung tissue were representative. It was possible to visualize intact structures, including bronchioles and alveoli. The activity of SOD and GSH-Px decreased thirty minutes after the biopsy. There were no changes in GSH-t and TBARS concentrations after the procedure. The percutaneous transthoracic lung... (Complete abstract click electronic access below)

Submitted by Amanda Nonato null (amandanonato@zootecnista.com.br) on 2017-05-04T18:17:51Z No. of bitstreams: 1 Tese Amanda Nonato.pdf: 1885961 bytes, checksum: 3b20e5e85bd1ef0f045b36418344a412 (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-05-05T13:29:43Z (GMT) No. of bitstreams: 1 nonato_a_dr_jabo.pdf: 1885961 bytes, checksum: 3b20e5e85bd1ef0f045b36418344a412 (MD5) / Made available in DSpace on 2017-05-05T13:29:43Z (GMT). No. of bitstreams: 1 nonato_a_dr_jabo.pdf: 1885961 bytes, checksum: 3b20e5e85bd1ef0f045b36418344a412 (MD5) Previous issue date: 2017-03-31 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A arginina é um aminoácido nutricionalmente essencial, importante na gestação de mamíferos, pois aumenta a sobrevivência, crescimento e desenvolvimento de embriões, fetos e neonatos. A arginina pode ser utilizada na síntese de poliaminas por uma via metabólica clássica, porém estudos recentes revelaram uma importante via alternativa. Todavia, ambas foram pouco investigadas. Nesse contexto, estudos são necessários a fim de identificar a ocorrência das vias metabólicas da arginina nos tecidos reprodutivos durante a gestação de ovelhas. Assim o presente estudo teve como objetivos: a) avaliar a expressão de genes das vias metabólicas da arginina em conceptos durante o período de peri-implantação em ovelhas; b) identificar a localização das proteínas das vias metabólicas da arginina em conceptos e no endométrio durante o período de peri-implantação em ovelhas; e c) identificar a localização das proteínas das vias metabólicas da arginina em placentomas em ovelhas. Para tanto, ovelhas da raça Rambouillet (n=72) foram sincronizadas e após detecção do estro por machos vasectomizados, foram cobertas por machos da raça Suffolk com fertilidade comprovada. No experimento 1, as ovelhas (n=20) foram histerectomizadas aos 13, 14, 15 ou 16 dias de gestação, e os fragmentos de conceptos coletados foram congelados em nitrogênio líquido a fim de se estudar a expressão gênica. No experimento 2, ovelhas (n=28) também foram histerectomizadas aos 13, 14, 15 ou 16 dias de gestação e, secções de conceptos e de endométrio, foram fixados a fim de avaliar a localização de proteínas por imunohistoquímica. No experimento 3, ovelhas (n=24) foram histerectomizadas aos 40, 60, 80, 100, 120 ou 140 dias de gestação e placentomas foram coletados e fixados para posterior análise imunohistoquímica. Os dados de expressão gênica foram analisados utilizando o procedimento GLM do SAS e as diferenças na intensidade da imunomarcação foram avaliadas com o software R, foi realizada a análise de variância (ANOVA). O nível de significância considerado foi <0,05. A expressão dos genes ADC e AGMAT (via alternativa), não foi alterada ao longos dos dias de gestação, e o mesmo ocorreu para o gene ARG1 (via clássica). O gene ODC1 (via clássica) apresentou maior expressão aos 13 dias de gestação (P=0,06). A análise imunohistoquímica identificou as proteínas ARG1, ODC1, ADC e AGMAT nos diferentes tecidos e dias de gestação, evidenciando a importância da ambas vias durante a gestação. A presença das proteínas no epitélio luminal do endométrio aumentou (p=0,038) após os 13 dias de gestação e foi significativamente maior aos 16 dias, sugerindo que essas proteínas são requeridas em maiores concentrações para o processo de alongamento do concepto e implantação. No epitélio glandular, a presença das proteínas não foi alterada (p=0,64) ao longo dos dias estudados. Não foi observada diferença (p=0,472) entre a identificação das proteínas no trofectoderma dos conceptos ao longo dos dias. Nos placentomas, a presença das proteínas foi menor (p<0,01) aos 40 dias de gestação, quando comparadas com os dias 60, 120 e 140. Os resultados do presente estudo evidenciam a importância de ambas vias metabólicas da arginina, para produção de poliaminas, durante o período gestacional de ovelhas. / Arginine is a nutritionally essential amino acid, important for pregnancy in mammalian, to increase the survival, growth and development of embryos, fetuses and neonates. Arginine can be used in the synthesis of polyamines by a classical metabolic pathway, though recent studies revealed an important alternative pathway, however, both have been poorly investigated. In this context, studies are necessary in order to identify an occurrence of arginine metabolic pathways in the reproductive tissues during a pregnancy in ewes. The present study had as objectives: a) evaluate the gene expression in the metabolic pathways of arginine, in concepts during the pre-implantation period of pregnancy in ewes; b) to identify the localization of proteins, from the metabolic pathways of arginine, in concepts and uterine endometrium during the pre-implantation period of pregnancy in ewes; and c) to identify the localization of proteins, from the metabolic pathway of arginine, in placentomes during the pre-implantation period of pregnancy in ewes. Rambouillet ewes (n=72) were synchronized and after estrus detection by vasectomized males, ewes were mated by Suffolk males with proven fertility. In experiment 1, ewes (n=20) were hysterectomized at days 13, 14, 15 or 16 of pregnancy, and the sections of concepts collected were frozen in liquid nitrogen in order to study gene expression. In the experiment 2, ewes (n=28) were also hysterectomized at days 13, 14, 15 or 16 of pregnancy, sections of conceptuses and uterine endometrium were fixed in order to identify the protein localization by immunohistochemical analysis. In experiment 3, ewes (n=24) were hysterectomized on days 40, 60, 80, 100, 120 or 140 days of pregnancy, placentomes were collected and fixed for immunohistochemical analysis. Data analysis of gene expression was analyzed using GLM procedure of SAS and differences in the intensity of the immunostaining were evaluated using software R, analysis of variance (ANOVA) was performed. Significance level was declared < 0.05. The expression of ADC and AGMAT genes (alternative pathway) did not alter across days of pregnancy, and the same happened with ARG1 gene (classic pathway). The ODC1 gene (classic pathway) showed greater expression at 13 days of pregnancy (P=0.06). The immunohistochemical analysis identified ARG1, ODC1, ADC and AGMAT proteins in all tissues and days of pregnancy studied, evidencing the relevance of both pathways during pregnancy. The identification of proteins on lumen epithelia from endometrium improved (p=0.038) after Day 13 of pregnancy and it was significantly greater on Day 16, suggesting that these proteins are required in higher concentrations for conceptuses elongation and implantation. On glandular epithelia, the proteins identification did not change (p=0.64) across days of pregnancy. No difference was observed (p=0.472) between the proteins identification on trofectoderm across days. On placentomes, the proteins identification was lower (p<0.01) on Day 40 of pregnancy compared with Days 60, 120 and 140. The results of the experiment indicate that relevance of both metabolic pathways of arginine to polyamines production during the pregnancy period in ewes.

Made available in DSpace on 2014-06-11T19:29:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-06-26Bitstream added on 2014-06-13T18:58:59Z : No. of bitstreams: 1 rodello_l_me_botfmvz.pdf: 226950 bytes, checksum: b009f4db5df84afd8c9c3af24e35fe7b (MD5) / Universidade Estadual Paulista (UNESP) / Objetivou-se avaliar e validar o sistema automatizado de refrigeração e congelação de fabricação nacional TK 3000, comparativamente ao sistema de refrigeração e de congelação pelo método geladeira/vapor de nitrogênio líqüido. Foram utilizados dez carneiros da raça Santa Inês sendo colhidos, por meio de vagina artificial, três ejaculados de cada reprodutor. Após as avaliações do ejaculado, o sêmen foi diluído em meio para congelação Glicina-Gema-Leite e envasado em palhetas de 0,25 Ml com 100 x 106 espermatozóides totais/doses, sob temperatura de 32° C. As amostras foram refrigeradas em geladeira e/ou com controle automatizado e posteriormente congeladas em vapor de nitrogênio líqüido e/ou com controle automatizado. Os resultados foram similares (P<0,05) entre a refrigeração em geladeira e a refrigeração com controle automatizados em todos os parâmetros avaliados pós descongelação e ao final do teste de exaustão, o mesmo observou-se entre a refrigeração em geladeira e congelação em vapor de nitrogênio líqüido e refrigeração e congelação com controle automatizado e as combinações refrigeração em geladeira e congelação com controle automatizado e refrigeração com controle automatizado e congelação em vapor de nitrogênio líqüido. Concluí-se que o sistema automatizado TK 3000 é adequado a criopreservação do sêmen ovino, com qualidade equivalente àquela processada no sistema geladeira/vapor de nitrogênio líqüido. / The objectives of the present study were to validate and evaluate a programmed automatic system for freezing and cooling, using a national equipment TK 3000, compared to a method using refrigerator and liquid nitrogen for the same procedures. Ten rams from Santa Ines breed were submitted to semen collection by artificial vagina, with a total of three ejaculates from each sire. After the ejaculate evaluation, semen was diluted in a freezing extender Egg- Glycine - Milk and stored in a 0.25 mL straws with a total of 100 x 10 6 sperm per dose at 32° C. The samples were cooled inside a refrigerator and /or utilizing an automatic programmed system, being then frozen in liquid nitrogen vapor and/ or using automatic control. The results were simmilar (P< 0.05) between samples submitted to cooling using refrigerator and automatic control for all analyzed parameters after thawing and at the end of the resistant test. The same was observed among samples submitted to cooling in refrigerator followed by liquid nitrogen vapor for freezing, samples cooled and frozen both using an automatic control, samples processed by the combinations of cooling in refrigerator followed by freezing with an automatic control, and samples cooled with automatic control followed by freezing in liquid nitrogen vapor. In conclusion, the TK 3000 is an adequate automatic system for ovine semen cryopreservation and it provides a quality which is equivalent to semen processed in refrigerator/ liquid nitrogen vapor system.

Orientador: Sony Dimas Bicudo / Banca: Carlos Antonio de Miranda Bomfim / Banca: André Luis Rios Rodrigues / Resumo: Objetivou-se avaliar e validar o sistema automatizado de refrigeração e congelação de fabricação nacional TK 3000, comparativamente ao sistema de refrigeração e de congelação pelo método geladeira/vapor de nitrogênio líqüido. Foram utilizados dez carneiros da raça Santa Inês sendo colhidos, por meio de vagina artificial, três ejaculados de cada reprodutor. Após as avaliações do ejaculado, o sêmen foi diluído em meio para congelação Glicina-Gema-Leite e envasado em palhetas de 0,25 Ml com 100 x 106 espermatozóides totais/doses, sob temperatura de 32° C. As amostras foram refrigeradas em geladeira e/ou com controle automatizado e posteriormente congeladas em vapor de nitrogênio líqüido e/ou com controle automatizado. Os resultados foram similares (P<0,05) entre a refrigeração em geladeira e a refrigeração com controle automatizados em todos os parâmetros avaliados pós descongelação e ao final do teste de exaustão, o mesmo observou-se entre a refrigeração em geladeira e congelação em vapor de nitrogênio líqüido e refrigeração e congelação com controle automatizado e as combinações refrigeração em geladeira e congelação com controle automatizado e refrigeração com controle automatizado e congelação em vapor de nitrogênio líqüido. Concluí-se que o sistema automatizado TK 3000 é adequado a criopreservação do sêmen ovino, com qualidade equivalente àquela processada no sistema geladeira/vapor de nitrogênio líqüido. / Abstract: The objectives of the present study were to validate and evaluate a programmed automatic system for freezing and cooling, using a national equipment TK 3000, compared to a method using refrigerator and liquid nitrogen for the same procedures. Ten rams from Santa Ines breed were submitted to semen collection by artificial vagina, with a total of three ejaculates from each sire. After the ejaculate evaluation, semen was diluted in a freezing extender Egg- Glycine - Milk and stored in a 0.25 mL straws with a total of 100 x 10 6 sperm per dose at 32° C. The samples were cooled inside a refrigerator and /or utilizing an automatic programmed system, being then frozen in liquid nitrogen vapor and/ or using automatic control. The results were simmilar (P< 0.05) between samples submitted to cooling using refrigerator and automatic control for all analyzed parameters after thawing and at the end of the resistant test. The same was observed among samples submitted to cooling in refrigerator followed by liquid nitrogen vapor for freezing, samples cooled and frozen both using an automatic control, samples processed by the combinations of cooling in refrigerator followed by freezing with an automatic control, and samples cooled with automatic control followed by freezing in liquid nitrogen vapor. In conclusion, the TK 3000 is an adequate automatic system for ovine semen cryopreservation and it provides a quality which is equivalent to semen processed in refrigerator/ liquid nitrogen vapor system. / Mestre

Orientador: Euclides Braga Malheiros / Coorientador: Ana Claudia Riggieri / Banca: Cledson Augusto Garcia / Banca: Elisabeth Criscuolo Urbinati / Resumo: Avaliou-se o efeito da intensidade de pastejo com e sem suplementação no perfil metabólico e indicadores de estresse em 64 cordeiros mestiços Santa Inês (15±3,31). O experimento foi avaliado em medidas repetidas no tempo em um delineamento inteiramente casualizado em esquema fatorial 4 x 2, sendo quatro intensidades de pastejo (IP) (0,8; 1,4; 2,0 e 2,6 de índice de área foliar residual) e grupos suplementados (SP) ou não (NSP). Foram realizadas coletas de sangue a cada 21 dias totalizando seis durante o período experimental. Foram mensurados a hemoglobina, creatinina, uréia, proteína total, albumina, globulina, glicose, ácidos graxos não esterificados (AGNE), beta-hidroxibutirato, colesterol, cálcio (Ca), fósforo (P), magnésio (Mg), relação neutrófilos:linfócitos (N:L) e cortisol plasmático. Nenhum dos indicadores sanguíneos do metabolismo protéico foi afetado pela IP e SP (p>0,05), exceto a albumina que foi maior (p<0,05) nos grupos que receberam suplemento. Os níveis da glicose foi maior (p<0,05) nos grupos suplementados e aumentou com a diminuição da IP (p<0,05). O AGNE apresentou níveis elevados nos grupos NSP (p<0,05) e reduziram com a diminuição da IP (p<0,05). O beta-hidroxibutirato foi modificado apenas pelo SP que foi maior (p<0,05) nos grupos que receberam suplemento. Os teores de Ca apresentaram-se crescentes com o decréscimo da IP (p<0,05). A concentração de P foi maior nos grupos SP (p<0,05). A N:L foi maior nos grupos NSP (p<0,05) e aumentou com o acréscimo da IP (p<0,05). O nível plasmático de cortisol foi maior nos grupos NSP (p<0,05). Esta caracterização metabólica mostrou que a IP e SP modificou o perfil metabólico e os indicadores sanguíneos de estresse de cordeiros em terminação / Abstract: Evaluated the effect of grazing intensity with supplementation or nonsupplementation in the metabolic profile and stress indicator in sixty four crossbred Santa Inês lambs (15,40±2,31 kg). The trial was evaluated in repeated measurement in a completely randomized design arranged 4 x 2 factorial in which grazing intensity (GI) (0.8 vs. 1.4 vs. 2.0 vs. 2.6 residual leaf area index), supplementation (SP) (nonsupplementation vs. supplementation) were the main effects to be investigated. Six sample of blood were taken during, every 21-days from the jugular vein. The hemoglobin, creatinine, urea, total protein, albumin, globulins, glucose, nonesterified fatty acids (NEFA), β-hydroxybutyrate, cholesterol, calcium (Ca), phosphorus (P), magnesium (Mg), neutrophil: lymphocyte rate (N:L) and cortisol were all measured. No indicator protein metabolism was not affected for GI and SP (p>0,05), except albumin, which was high (p<0,05) in supplemented groups. The glucose levels was high in supplemented groups (p<0,05) and increased with a decrease of GI (p<0,05). The NEFA showed high levels in non supplemented groups (p<0,05) and increased with a decrease of GI (p<0,05). The β-hydroxybutyrate levels was modified only for SP (p<0,05) and supplemented groups showed greater concentration. The Ca levels showed increase with decrease of GI (p<0,05) in both supplemented and non supplemented groups. The phosphorus levels was high in supplemented groups (p<0,05). The N:L rate was modified for GI (p<0,05) showed increased levels with the increase of GI and presented high in non supplemented groups (p<0,05). The levels of cortisol was modified for supplementation only (p<0,05). This metabolic characterization showed clearly the grazing intensity and supplementation modified the metabolic profile and stress indicator in finishing lambs / Mestre

Orientador: Ana Silvia Alves Meira Tavares Moura / Resumo: O objetivo do presente trabalho foi avaliar o efeito da adição de óleo de linhaça (OL) ou farelo de linhaça (FL) na dieta de ovelhas da raça Bergamácia em lactação, sobre o teor de aminas biogênicas e polifenóis no leite. Foram utilizadas 70 ovelhas distribuídas aleatoriamente entre os tratamentos. As dietas experimentais foram: Controle (CT) – concentrado sem adição de suplementação lipídica; Óleo de linhaça (OL) – concentrado com adição de 3% de OL (%MS); e Farelo de linhaça (FL) – concentrado com adição de 15% de FL (%MS). Durante todo o período experimental, os animais foram mantidos confinados em baias coletivas e recebiam dieta composta por 60% de silagem de milho e 40% de concentrado referente a cada tratamento. As ovelhas foram ordenhadas uma vez ao dia, no período da manhã, e tiveram suas produções controladas diariamente. As amostras de leite para a avaliação das aminas biogênicas foram colhidas a cada 14 dias a partir do primeiro dia de ordenha e as amostras de leite para análise dos polifenóis foram colhidas a cada 14 dias a partir do quinto dia pós-parto. O experimento foi conduzido no delineamento inteiramente casualizado e os dados avaliados por análise de variância (P< 0,05). Não foi encontrada a presença de cadaverina no leite e a serotonina foi a amina predominante no leite de ovelhas em todos os tratamentos. O tratamento controle apresentou maiores teores de espermina (0,300 mg/100g) entretanto, o tratamento com óleo de linhaça apresentou menores teores de ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The aim in this study was to determine the effects of feeding diets with linseed oil (OL) or flaxseed meal (FM) of Bergamasca ewes on to the content of biogenic amines and phenols in milk.Seventy ewes were distributed in three groups: Control (CT) - no lipids; Linseed (L) – with addition of 3% of OL (DM basis); and Flaxseed meal (F) – with addition of 15% of FM (DM basis). Throughout the experiment, the ewes remained confined in collective pens and were fed a diet containing 60% corn silage and 40% concentrate according to each treatment. They were mechanically milked once a day and had their productions controlled. The milk samples for biogenic amines analysis were collected every 14 days from the first milking day and the milk samples for phenols analysis were collected every 14 days from the 5th day postpartum until de end of the lactation. The experiment was conducted in a random customized design and the data evaluated by analysis of variance (P< 0,05). Cadaverine was not found and the serotonine was the predominant amine in the milk from ewes in all treatments. The control treatment showed high contents of espermine (0,300 mg/100g), however the OL had lower contents of histamine (0,0049 mg/100g). The espermidine was the biogenic amine more correlated with greater numbers of amines. There was no difference between treatments for polyphenols contents in milk, however the treatment with oil it was more efficient. / Doutor