Níveis elevados de proteína bruta (PB) na dieta de ruminantes têm sido associado com comprometimento na fertilidade devido, principalmente, a elevação da concentração do nitrogênio uréico plasmático (NUP). Objetivou-se no presente estudo avaliar o efeito do fornecimento de dietas contendo nitrogênio não-protéico (NNP) sem prévia adaptação, durante curto espaço de tempo e em diferentes fases do ciclo estral, a fêmeas bovinas superovuladas quanto à produção, qualidade e ao grau de desenvolvimento de embriões recuperados. Sessenta e oito vacas da raça Nelore com escore corporal de 7,56 e peso médio de 557,6 kg foram distribuídas em três tratamentos: controle (C), uréia antes do dia 0 (UA; fornecimento de dieta com uréia do dia -5 ao dia 0) e uréia depois do dia 0 (UD; fornecimento de dieta com uréia do dia 0 ao dia 5). As vacas foram mantidas em piquetes com pastagem de Brachiaria brizantha cv. Marandu e receberam diariamente, 3,0 kg/animal de concentrado durante 16 dias. Foram formulados dois concentrados, sendo que as dietas totais (concentrado + consumo estimado de pastagem) continham 12,0% (dieta controle) e 14,6% de PB (dieta com NNP). A diferença na PB entre as dietas foi o acréscimo de 100g/vaca/dia de uréia. Os animais foram sincronizados, superovulados e inseminados. Sete dias (dia 7) após a inseminação (dia 0), realizou-se a colheita e análise dos embriões quanto à qualidade a ao grau de desenvolvimento. Amostras de sangue foram coletadas nos dias -5, 0 e 5 para mensurações das concentrações de NUP, glicose, insulina e progesterona. Houve efeito significativo dos tratamentos sobre a concentração médias de NUP no dia -5 (C = 16,18ab, UA = 17,72a e UD = 14,47b mg/dL; P = 0,0007), no dia 0 (C = 18,95b, UA = 22,71a e UD = 21,15 ab mg/dL; P = 0,0010) e no dia 5 (C = 20,93a, UA = 19,08b e UD = 21,51a mg/dL; P = 0,0022). Quanto às concentrações de glicose, insulina e progesterona não houve efeito dos tratamentos. Não houve efeito significativo nem da inclusão da uréia e nem do momento da inclusão da uréia sobre o total de estruturas recuperadas, total de estruturas fecundadas e de embriões viáveis. Porém, houve efeito do momento de inclusão da uréia sobre a porcentagem de estruturas fecundadas em relação ao total de estruturas recuperadas (UA = 75,3 vs. UD = 50,7%) e sobre a porcentagem de embriões viáveis em relação ao total de estruturas recuperadas (UA = 73,2 vs. UD = 39,4%). O tratamento UD acarretou em menor capacidade de fecundação do oócito em relação ao tratamento UA, havendo redução de aproximadamente 32,7% na proporção de oócitos fecundados sobre o total de estruturas recuperadas deste tratamento. / High levels of crude protein (CP) in ruminant diets were associated with detrimental effect on fertility, mainly, due to the rise of plasmatic urea nitrogen (PUN) concentration. The aim of this study was to verify the effects of short term non-protein nitrogen feeding at different periods of the oestrus cycle in superovulated cows without previous adaptation on yield, quality and development degree of recovered embryos. Sixty eight Nelore cows with body condition score of 7.56 and body weight of 557.6 kg were divided in three different groups: control (C), urea before day 0 (UB; urea supply from day -5 to day 0) and urea after day 0 (UA; urea supply from day 0 to day 5). Animals were grazing in Brachiaria brizantha cv. Marandu pasture and received 3.0 kg/animal/day of concentrate during 16 days. Two concentrates were formulated and the total diets (concentrate + estimate forage intake) had 12.0% (control diet) and 14.6% of CP (NPN diet). The difference in CP between diets was the addition of 100g/cow/day of urea. Animals were synchronized, superovulated and inseminated. The embryos were collected seven days (day 7) after insemination (day 0) and quality and development degree were also evaluated. Blood samples were collected on day -5, 0 and 5 for measurement of PUN, glucose, insulin and progesterone. There was significant effect of treatments on average PUN concentration at day -5 (C = 16.18ab, UB = 17.72a and UA = 14.47b mg/dL; P = 0.0007), at day 0 (C = 18.95b, UB = 22.71a and UA = 21.15ab mg/dL; P = 0.0010) and at day 5 (C = 20.93a, UB = 19.08b and UA = 21.51a mg/dL; P = 0.0022). For glucose, insulin and progesterone there was no effect of treatments. It was observed neither effect of urea inclusion nor the moment of urea inclusion in diet on the total number of recovered structures, total number of fertilized structures and total number of viable embryos. However, it was observed effect of the moment of urea inclusion on the rate of fertilized structures in relation to the total number of recovered structures (UB = 75.3 vs. UA = 50.7%) and on the rate of viable embryos in relation to the total number of recovered structures (UB = 73.2 vs. UA = 39.4%). The UA treatment lead to lowest fertilizing oocyte capacity in relation to UB treatment with an approximate reduction of 32.7% on oocytes fertilized proportion over the total recovered structures in this treatment.

Made available in DSpace on 2016-12-08T16:24:21Z (GMT). No. of bitstreams: 1 PGCV05MA012.pdf: 393991 bytes, checksum: 8755a862d62c91605bfe0f6eb41d8e56 (MD5) Previous issue date: 2005-08-09 / The effects of the age on the 120th, 180th and 240th days, concerning the nourishing behavior and the ponderal performance of thirteen Texel lambs were studied.The research was conducted in the Sector of Sheep Raising of CAV/UDESC, Lages, SC. The lambs were weaned with 60 days of age and fed during the day, with water and mineral salt ad libitum, in enclosures of 0,8ha, with red clover manured pasture (Trifolium pratense L.) and white clover (Trifolium repens L.) and at night they were sheltered in collective stalls and supplemented with alfalfa hay (Medicago sativa L.). The animals were wormed and vaccinated against enteroxotemy. Samples of veiny blood of 10mL were collected from the jugular vein, and in 3mL anti-coagulant Ethylenediaminotetraceticodissodic was added, in 10% concentration, by means of 0,1mL in each 5mL blood for the hematologic survey, while the remaining 7mL were coagulated, and its serum was used for the biochemical dosing. From the ear 1mL of artery blood was collected in order to establish the hemogasometric values. The average hematologic values which were gotten for the studied ages were, respectively, hematocrit or globular volume (VG) 32,50; 33,06 and 35,80%; the average corpuscular hemoglobinic concentration (CHCM) was 36,73; 37,02 and 38,09g/dL; erythrocyte 9,86x106; 10,69 x106 and 9,04 x106 per &#956;L; hemoglobin (Hb.) 11,92; 12,60 and 12,97 g/dL; average corpuscular hemoglobin (HCM) 12,12; 12,16 and 14,1 pg. The plasmatic biochemical values referring to the researched ages were, respectively, glucose 79,09; 81,57 and 82,96 mg/dL; cholesterol of 42,37; 45,94 and 47,51 mg/dL; magnesium pf 1,90; 1,81 and 1,87 mg/dL; phosphorus of 7,47; 7,34 and 7,19 mg/dL; total calcium 11,37; 11,18 and 11,15 mg/dL; sodium of 155,11; 150,73 and 154,18 mEq/L; potassium of 4,86; 4,76 and 4,91 mEq/L. The hematologic values, according to sex, were respectively, hematocrit or globular volume (VG) 30,35 and 30,03%; average cospuscular hemoglobinic concentration (CHCM) 37,69 and 36,95 g/dL; erythrocytes of 9,03x106 and 10,25x106 per &#956;L; hemoglobin (Hb) 11,42 and 12,96 g/dL; average corpuscular volume (VCM) of 34,29 and 34,28 fL; leucocytes of 10,45 and 13,03 cel./ &#956;L; average cospuscular hemoglobin (HCM) 12,91 and 12,67 pg. The plasmatic biochemical values according to sex, were respectively, glucose of 82,37 snf 81,48 mf/dL; cholesterol of 44,03 and 46,44 mg/dL; magnesium of 1,87 and 1,85 mg/dL; phosphorus of 6,72 and 7,95 mg/dL; total calcium 11,13 and 11,31 mg/dL; sodium of 153,31 and 153,37 mEq/L; potassium of 4,74 and 4,94 mEq/L. The average hemogasometric values, corrected for he corporal temperature of 38ºC, according to the ages of 120, 180 and 240 days were, respectively, pH 7,45; 7,44 and 7,44; PaCO2 of 34,88, 34,65; 31,94 mmHg; PaO2 81,86; 77,68; 78,52 mmHg and BE 1,08 -1,78 and 0,13mEq/L. For the sex variable it was registered, respectively as pH of 7,45 and 7,38; PaCO2 pf 37,71 and 31,00 mmHg and PaO2 of 75,29 and 83,42 mmHg and BE 1,08 and 1,04 mEq/L. There was no meaningful difference (P<0,05) for the researched variables. The behaviorial data were obtained through visual observation, on the field, with interval of 5 minutes, during consecutive daily periods of 10 hours, in the experimental age bands, corresponding to the months from December 2003 to February and april 2004. Weighing was held at each 14 days, in a total of 15 trials. The scores of time spent in pasturing in the sun presented average values of 5,58; 5,00 and 1,70 h/day; the pasturing in the shadow consisted of average times of 1,70; 2,37 and 5,28 h/day xiii 14 and the rumination time in the shadow corresponded to 1,23; 1,88 and 1,77 h/day. The average values of daily weight gain, from birth to weaning and from weaning to the 240 days were, respectively, of 25,67 and 229,74 g/d. The data were analysed and delineation was entirely determined in factorial structure for sex and age. There was no effect (P<0,05) of interaction between age and sex, and from the sex factor to the analysed variables / Foram estudados os efeitos de idades aos 120, 180 e 240 dias, sobre o comportamento alimentar e o desempenho ponderal de treze cordeiros da raça Texel. O experimento foi realizado no Setor de Ovinocultura do CAV/UDESC em Lages, SC. Os cordeiros foram desmamados aos 60 dias e alimentados durante o dia, com água e sal mineral ad libitum, em piquete de 0,8 hectare com pastagem cultivada de trevo vermelho (Trifolium pratense L.) e trevo branco (Trifolium repens L.) e à noite eram estabulados em baias coletivas e suplementados com feno de alfafa (Medicago sativa L.). Os animais foram vermifugados e vacinados contra enterotoxemia. Foram obtidas amostras de 10 mL sangue venoso da veia jugular, sendo em 3 mL adicionado anticoagulante Etileno-diamino-tetracéticodissódico na concentração de 10%, utilizando-se 0,1mL em cada 5mL de sangue para os exames hematológicos e os restantes 7 mL foram deixados para que coagulasse e o soro do mesmo utilizado para as dosagens bioquímicas. Da artéria marginal da orelha, colheitou-se 1 mL de sangue para determinação dos valores hemogasométricos. Os valores hematológicos médios obtidos para as idades estudadas, foram, respectivamente,:hematócrito ou volume globular (VG) 32,50; 33,06 e 35,80%; concentração hemoglobínica corpuscular média (CHCM) 36,73; 37,02 e 38,09 g/dL; hemácias 9,86 × 106; 10,69 × 106 e 9,04 × 106 por &#956;L; hemoglobina (Hb) 11,92; 12,60 e 12,97 g/dL; volume corpuscular médio (VCM) 33,01; 32,90 e 36,81 fL; leucócitos 12.600; 10.730; 11.900 cel./&#956;L; hemoglobina corpuscular média (HCM) 12,12; 12,16 e 14,10 pg. Os valores bioquímicos plasmáticos referentes as idades estudadas foram, respectivamente,:glicose 79,09; 81,57 e 82,96 mg/dL; colesterol de 42,37; 45,94 e 47,51 mg/dL; magnésio de 1,90; 1,81 e 1,87 mg/dL; fósforo de 7,47; 7,34 e 7,19 mg/dL; cálcio total 11,37; 11,18 e 11,15 mg/dL; sódio de 155,11; 150,73 e 154,18 mEq/L; potássio de 4,86; 4,76 e 4,91 mEq/L. Os valores hematológicos, segundo o sexo, foram respectivamente, hematócrito ou volume globular (VG) 30.35 e 35,06%; concentração hemoglobínica corpuscular média (CHCM) 37,69 e; 36,95 g/dL; hemácias de 9,03 × 106 e 10,25 × 106 por &#956;L; hemoglobina (Hb) 11,42 e 12,96 g/dL; volume corpuscular médio (VCM) de 34,29 e 34,28 fL leucócitos de 10,45 e 13,03 cel./&#956;L; hemoglobina corpuscular média (HCM) 12,91 e 12,67 pg. Os valores bioquímicos plasmáticos segundo o sexo foram, respectivamente, glicose de 82,37 e 81,48 mg/dL; colesterol de 44,03 e 46,44 mg/dL; magnésio de 1,87 e 1,85 mg/dL; fósforo de 6,72 e 7,95 mg/dL; cálcio total 11,13 e 11,31 mg/dL; sódio de 153,31 e 153,37 mEq/L; potássio de 4,74 e 4,94 mEq/L. Os valores hemogasométricos médios, corrigidos para temperatura corporal de 38ºC, segundo as idades de 120, 180 e 240 dias foram, respectivamente: pH 7,45; 7,44 e 7,44; PaCO2 de 34,88; 34,65; 31,94 mmHg; PaO2 81,86; 77,68; 78,52 mmHg e BE 1,17, -1,78 e 0,13 mEq/L Para a variável sexo macho e fêmea registrou-se, respectivamente, pH de 7,45 e 7,38; PaCO2 de 37,71 e 31,00 mmHg e PaO2 de 75,29 e 83,42 mmHg e BE 1,08 e 1,04 mEq/L. Não houve diferença significativa (P<0,05) para as variáveis estudadas. Os dados comportamentais foram obtidos por observação visual, a campo, a cada cinco minutos, por três períodos diários consecutivos de dez horas, nas faixas etárias experimentais, correspondendo aos meses de dezembro de 2003, fevereiro e abril de 2004. Efetuaram-se pesagens a cada quatorze dias, totalizando quinze pesagens. Os tempos despendidos em pastejo ao sol, apresentaram valores xi 12 médios de 5,58; 5,00 e 1,70 h/dia; em pastejo à sombra, tempos médios de 1,70; 2,37 e 5,28 h/dia e tempo de ruminação à sombra de 1,23; 1,88 e 1,77 h/dia. Os valores médios de ganho de peso diário, do nascimento ao desmame e do desmame aos 240 dias foram, respectivamente, de 257,67 e 229,74 g/d. Os dados foram analisados em delineamento inteiramente casualizado em estrutura fatorial para sexo e idade. Não houve efeito (P<0,05) de interação entre idade e sexo e do fator sexo para as variáveis analisadas

Resumo: O experimento foi conduzido no confinamento da Unesp/Botucatu, com o objetivo de avaliar a influência da suplementação das vitaminas D e E sobre o desempenho, características de carcaça e atributos de carne de bovinos jovens confinados. Foram utilizados 36 machos inteiros, 18 Nelore (NE) e 18 Canchim (CC); desmamados aos sete meses de idade, com peso vivo médio de 234,53 ± 22,15 e 248,13 ± 34,67 kg, respectivamente. Os animais foram confinados por 134 dias. Aos quarenta e sete dias pré-abate, nove animais NE e nove animais CC, foram suplementados com 1300 UI/vitamina E/dia e 10 dias antes do abate, os mesmos animais foram suplementados com 7,5 x 106 UI/vitamina D3/dia. No último dia da suplementação, foram colhidas amostras de sangue dos animais, para avaliar o cálcio plasmático. O pH da carcaça foi mensurado à zero e 24hs após o abate. Na desossa, foram colhidas amostras de Longissimus, entre a 12a e 13a costelas, para análise de força de cisalhamento, perdas por cocção, índice de fragmentação miofibrilar, lipídeos totais, concentração de vitamina D e E na carne e exposição sob condições simuladas de varejo. A cor e o pH da carne, foram mensurados diariamente nas amostras expostas. No primeiro e no último dia de exposição foram colhidas amostras da gordura subcutânea, para análise do perfil de ácidos graxos. Não houve efeito da suplementação de vitaminas D e E sobre o desempenho, características de carcaça e atributos da qualidade da carne, porém houve efeito para cálcio plasmático. A suplementação de vitaminas D e E, não foi efetiva para melhorar o desempenho, as características de carcaça e os atributos de qualidade da carne de bovinos jovens confinados. / Abstract: The experiment was conducted at Unesp/Botucatu feedlot, with the objective to evaluate the supplementation of vitamin D and E on performance, carcass characteristics and meat quality of feedlot bullocks. It was used 36 animals, 18 Nellore (NE) and 18 Canchim (CC), weaned at seven months and average initial body weight of 234.53 ± 22.15 and 248.13 ± 34.67 kg, respectively, fed for134 days. At the forty seventh day prior slaughter, nine NE and nine CC were supplemented with 1300 IU/vitamin E/day and 10 days prior slaughter, the same animals were supplemented with 7.5 x 106 IU/vitamin D3/day. At last day of supplementation, blood samples were taken to evaluate plasmatic calcium. Carcass pH was measured at 0 and 24hrs after slaughter. At boning process, Longissimus samples were collected between 12th and 13th ribs for sear force, cooking loss, miofibrilar fragmentation index, total lipids, vitamin D and E concentrations, and retail conditions storage simulation. Meat color and pH of the samples were daily measured. Fatty acid profile samples were collected at first and last day of retail condition storage simulation. There was no effect of vitamin D and E on feedlot performance, carcass characteristics and meat quality, although plasmatic calcium effect was observed. In conclusion, vitamin D and E were not effective to enhance feedlot performance, carcass characteristics and meat quality of feedlot bullocks. / Orientador: Mário De Beni Arrigoni / Coorientador: Cyntia Ludovico / Banca: Angélica Simone do Cravo Pereira / Banca: Saulo da Luz e Silva / Mestre

L’administration de fer dextran à 10 mg/kg intramusculaire (IM) est un traitement empirique couramment recommandé en médecine aviaire lors d’hémorragie ou d’anémie. L’objectif principal de cette étude était d’évaluer les effets de ce traitement sur l’anémie chez les oiseaux de proie. Deux types d’individus ont été utilisés : des crécerelles d’Amérique (Falco sparverius) où une anémie par perte de sang externe aiguë a été créée (deux phlébotomies de 20-40 % du volume sanguin total à un intervalle de 6 h) et des oiseaux de proie sauvages de différentes espèces souffrant d’anémies diverses. L’ensemble des oiseaux a été subdivisé aléatoirement en groupe traitement (fer dextran 10 mg/kg IM) et contrôle (NaCl 0,9% IM). Un suivi dans le temps a été réalisé afin d’étudier leur récupération de l’anémie, la présence d’effets secondaires au traitement et l’impact d’une administration de fer sur ces réserves. Aucune différence significative n’a été observée entre les deux groupes en ce qui concerne les signes cliniques, l’hématocrite, le pourcentage des polychromatophiles/réticulocytes, la densité cellulaire et le fer de la moelle osseuse, la créatine kinase et le fer plasmatique. La majorité des crécerelles ont présenté une myosite au site d’injection du fer. Nos résultats suggèrent qu’une administration de 10 mg/kg de fer dextran IM n’a pas d’effet sur l’érythropoïèse des rapaces souffrant d’anémie par perte de sang externe aiguë, qu’elle provoque une légère inflammation au site d’injection et qu’elle n’influence pas les réserves de fer. Le comptage des réticulocytes en anneau et des polychromatophiles semble être deux méthodes équivalentes. / A 10 mg/kg intramuscular (IM) administration of iron dextran is a common empirical treatment recommended in avian medicine for hemorrhage and anemia. The purpose of this study was to evaluate the effects of this treatment on anemia in birds of prey. Two kinds of specimen were used: the American kestrel (Falco sparverius) where an acute external blood loss anemia was created (with two phlebotomies of 20-40 % of the total blood volume at 6 hours interval) and other various species of wild birds of prey suffering from different types of anemia. All subjects were randomized into a treatment (iron dextran 10 mg/kg IM) or a control (NaCl 0,9 % IM) group. Monitoring was carried out to evaluate the evolution of the anemia, presence of side effects and impact of an iron administration on their iron reserve. No significant differences were observed between the two treatment groups for clinical signs, packed cell volume, the percentage of reticulocytes/polychromatophilic erythrocytes, bone marrow cellularity and iron, plasmatic iron and creatine kinase. Most kestrels had a myositis at the iron injection site. Our results suggest that an IM injection of 10 mg/kg iron dextran has no effect on raptor erythropoiesis after an acute external blood loss anemia, that it has no effect on iron reserve, and that it can cause mild inflammation at the injection site. The polychromatophilic erythrocytes and the reticulocytes ring form count were two equivalent methods.

A homocisteína plasmática é considerada marcador de risco para doenças cardiovasculares e sua associação com a hipertensão arterial essencial parece ser importante no agravamento desta doença. O exercício físico tem mostrado eficácia na redução da pressão arterial e o ácido fólico suplementar a dieta como a melhor conduta para a redução da homocisteinemia. O objetivo deste trabalho foi verificar o efeito da suplementação de ácido fólico e/ou do exercício físico sobre as concentrações plasmáticas de homocisteína em hipertensos essenciais medicados. Para tanto foram estudados 69 hipertensos (57&#177;10 anos), sendo 22 do sexo masculino e 47 do sexo feminino, divididos em dois grupos: grupo 1 medicado com diurético e grupo 2 medicado sem diurético, sendo o tiazídico o diurético mais utilizado (80%). Foram realizadas avaliações médica, antropométrica, dietética, pressórica, bioquímica e o teste de aptidão cardiorrespiratória. Os indivíduos receberam suplementação com 500 &#181;g/dia de ácido fólico, em estudo do tipo cruzado, e foram submetidos a treinamento com exercícios físicos supervisionados, sendo reavaliados a cada dois meses até o final do estudo. Os grupos foram homogêneos em relação a todas as variáveis estudadas exceto a homocisteína plasmática que foi maior significativamente no grupo 1. Não houve diferença significativa entre os sexos para todas as variáveis estudadas. Os hipertensos eram em sua maioria (62%) não controlados e portadores de síndrome metabólica (77%). O exercício físico apresentou efeito benéfico na reclassificação dos hipertensos leves e moderados para a classe de limítrofes sem normalizar a pressão arterial. Não houve efeito do exercício na composição corporal e na homocisteinemia. A suplementação de ácido fólico reduziu a hiperhomocisteinemia em 11% no grupo 1 e 19% no grupo 2 frente a aumentos similares na folacemia. Em relação à pressão arterial houve normalização da homocisteinemia apenas nos hipertensos com pressão arterial normalizada. Os hipertensos limítrofes e leves apresentaram reclassificação da hiperhomocisteinemia limítrofe para moderada. A presença de diurético potencializou a normalização da pressão pelo exercício físico e indiretamente influenciou, nesses casos, a normalização da homocisteína. Entretanto, a maior redução das concentrações médias de homocisteína ocorreu coma oferta de ácido fólico na ausência de diurético. Desta forma, recomenda-se a adequação dietética do folato como adjuvante terapêutico da hipertensão arterial associadamente ao exercício físico e/ou medicamentos / The plasma homocysteine is considered to be a risk indicator for cardiovascular diseases and its association with essential hypertension seems to be important to the worsening of this disease. Physical exercises are efficient to reduce blood pressure and the dietary supplements of folic acid are considered the best way to decrease homocysteine. The aim of this study was investigate the effect of acid folic supplementation and physical exercises on plasmatic levels of homocysteine in essential hypertensive medicated individuals. In order to that, 69 hypertensive subjects were studied (22 men and 47 women) divided in two groups: group 1 medicated with diuretic and group 2 medicated without diuretic. Medical, anthropometrical, dietetic, blood pressure, biochemistry and cardio-respiratory capacity evaluations were performed. These individuals received 500 &#181;g/dia of folic acid supplementation in a cross-over design and were submitted to daily supervisioned physical exercise. They were evaluated every two months until the end of the study. The groups were similar for all variables except for the homocysteine which was significantly greater in the group 1. There were no difference for the gender to all variables. The high blood pressure was not controlled (62%) and 77% of the patients showed metabolic syndrome. The physical exercise showed benefit on the reclassification of slight and moderated hypertensives to border-line hypertension, however, it did not normalize the blood pressure. There were no effects on the body composition and on homocysteine values . Folic acid supplementation decreased 11% of hyperhomocysteinemia in the group 1 and 19% in group 2 face to similar increases in folate. Regarding to high blood pressure subjects the homocysteine normalization occurred only in hypertensive subjects with normalized blood pressure. The border and slightly hypertensive individuals had border-line hyperhomocysteine wich were reclassificated to moderated c1ass. The use of diuretic enhanced blood pressure normalization with the physical exercise and indirectly influenced the plasma homocysteine normalization. However the greater decreasing mean homocysteine concentration occurred with folic acid but in the diuretic absence. So we strong recommend an adequate folate intake as a terapheutical adjuvant to arterial hypertension treatments either with physical exercise and/or antihipertensive drugs.

L’administration de fer dextran à 10 mg/kg intramusculaire (IM) est un traitement empirique couramment recommandé en médecine aviaire lors d’hémorragie ou d’anémie. L’objectif principal de cette étude était d’évaluer les effets de ce traitement sur l’anémie chez les oiseaux de proie. Deux types d’individus ont été utilisés : des crécerelles d’Amérique (Falco sparverius) où une anémie par perte de sang externe aiguë a été créée (deux phlébotomies de 20-40 % du volume sanguin total à un intervalle de 6 h) et des oiseaux de proie sauvages de différentes espèces souffrant d’anémies diverses. L’ensemble des oiseaux a été subdivisé aléatoirement en groupe traitement (fer dextran 10 mg/kg IM) et contrôle (NaCl 0,9% IM). Un suivi dans le temps a été réalisé afin d’étudier leur récupération de l’anémie, la présence d’effets secondaires au traitement et l’impact d’une administration de fer sur ces réserves. Aucune différence significative n’a été observée entre les deux groupes en ce qui concerne les signes cliniques, l’hématocrite, le pourcentage des polychromatophiles/réticulocytes, la densité cellulaire et le fer de la moelle osseuse, la créatine kinase et le fer plasmatique. La majorité des crécerelles ont présenté une myosite au site d’injection du fer. Nos résultats suggèrent qu’une administration de 10 mg/kg de fer dextran IM n’a pas d’effet sur l’érythropoïèse des rapaces souffrant d’anémie par perte de sang externe aiguë, qu’elle provoque une légère inflammation au site d’injection et qu’elle n’influence pas les réserves de fer. Le comptage des réticulocytes en anneau et des polychromatophiles semble être deux méthodes équivalentes. / A 10 mg/kg intramuscular (IM) administration of iron dextran is a common empirical treatment recommended in avian medicine for hemorrhage and anemia. The purpose of this study was to evaluate the effects of this treatment on anemia in birds of prey. Two kinds of specimen were used: the American kestrel (Falco sparverius) where an acute external blood loss anemia was created (with two phlebotomies of 20-40 % of the total blood volume at 6 hours interval) and other various species of wild birds of prey suffering from different types of anemia. All subjects were randomized into a treatment (iron dextran 10 mg/kg IM) or a control (NaCl 0,9 % IM) group. Monitoring was carried out to evaluate the evolution of the anemia, presence of side effects and impact of an iron administration on their iron reserve. No significant differences were observed between the two treatment groups for clinical signs, packed cell volume, the percentage of reticulocytes/polychromatophilic erythrocytes, bone marrow cellularity and iron, plasmatic iron and creatine kinase. Most kestrels had a myositis at the iron injection site. Our results suggest that an IM injection of 10 mg/kg iron dextran has no effect on raptor erythropoiesis after an acute external blood loss anemia, that it has no effect on iron reserve, and that it can cause mild inflammation at the injection site. The polychromatophilic erythrocytes and the reticulocytes ring form count were two equivalent methods.

Fundação de Amparo a Pesquisa do Estado de Minas Gerais / Leptospirosis is a zoonosis of global public health importance and can affect all groups of vertebrates, including reptiles. These animals can play a role in the epidemiological cycle of the disease spreading and keeping the causative agent in the environment. Leptospirosis is little known in snakes and this study was designed to cover the gaps in the occurrence of the disease in these animals. The occurrence of anti-Leptospira spp. antibodies in rattlesnakes Crotalus durissus collilineatus kept in captivity was evaluated and the most common serotypes in these animals were determined using the Microscopic Agglutination Test (MAT). Changes in concentrations of plasma biochemical constituents in positive snakes were also evaluated, depending on the antibody titre presented at MAT. Almost 90% of the rattlesnakes used in this study were positive to the serological test and the most common serotypes were Javanica, Andamana and Patoc. The most common titres were 25 and 50, but there were titres as high as 1600. Captive snakes can act as important sources of leptospira infection for humans and so it is critical that the disease is recognized and controlled. Although apparently healthy, alterations in biochemical parameters indicated inflammation and infection in animals with low antibody titres. Rattlesnakes with high titres showed signs of severe kidney damage and liver disease, which are common in leptospirosis. The biochemical assessment of positive MAT reptiles may be important to diagnose the infection in these animals. / A leptospirose é uma zoonose de importância em saúde pública global e pode acometer praticamente todos os grupos de vertebrados, inclusive répteis. Esses animais podem atuar no ciclo epidemiológico da doença mantendo e disseminando o agente causador no ambiente. A leptospirose é pouco conhecida em serpentes e, por isso, objetivou-se estudar a ocorrência da doença nesses animais. Para isso, foi avaliada a ocorrência de anticorpos anti-Leptospira spp. em Crotalus durissus collilineatus mantidas em cativeiro e as sorovariedades mais frequentes nesses animais, utilizando o teste de Soroaglutinação Microscópica (SAM). Também foram avaliadas as alterações nas concentrações de constituintes bioquímicos plasmáticos nas serpentes reagentes, relacionadas com o título de anticorpos apresentado na SAM. Quase 90% das cascavéis utilizadas nesse estudo foram reagentes no teste sorológico com triagem de 1:25 e os sorovares mais frequentes foram Javanica, Andamana e Patoc. Os títulos mais frequentes foram 25 e 50, mas houve títulos de até 1600. Apesar de aparentemente saudáveis, a avaliação dos parâmetros bioquímicos indicou inflamação e infecção mesmo nos animais com títulos de anticorpos considerados baixos para mamíferos (&#8804;50). As cascavéis com títulos altos apresentaram quadros mais graves que aquelas com títulos menores, sugestivos de lesão renal grave e indícios de doença hepática, que são lesões características da leptospirose. Serpentes de cativeiro podem atuar como fontes de infecção de leptospiras para humanos e outros animais e por isso, é fundamental que a doença seja conhecida e controlada em criadouros. A avaliação bioquímica de répteis reagentes no teste sorológico pode ser importante para o diagnóstico da infecção por Leptospira spp. nesses animais. / Mestre em Ciências Veterinárias

A homocisteína plasmática é considerada marcador de risco para doenças cardiovasculares e sua associação com a hipertensão arterial essencial parece ser importante no agravamento desta doença. O exercício físico tem mostrado eficácia na redução da pressão arterial e o ácido fólico suplementar a dieta como a melhor conduta para a redução da homocisteinemia. O objetivo deste trabalho foi verificar o efeito da suplementação de ácido fólico e/ou do exercício físico sobre as concentrações plasmáticas de homocisteína em hipertensos essenciais medicados. Para tanto foram estudados 69 hipertensos (57&#177;10 anos), sendo 22 do sexo masculino e 47 do sexo feminino, divididos em dois grupos: grupo 1 medicado com diurético e grupo 2 medicado sem diurético, sendo o tiazídico o diurético mais utilizado (80%). Foram realizadas avaliações médica, antropométrica, dietética, pressórica, bioquímica e o teste de aptidão cardiorrespiratória. Os indivíduos receberam suplementação com 500 &#181;g/dia de ácido fólico, em estudo do tipo cruzado, e foram submetidos a treinamento com exercícios físicos supervisionados, sendo reavaliados a cada dois meses até o final do estudo. Os grupos foram homogêneos em relação a todas as variáveis estudadas exceto a homocisteína plasmática que foi maior significativamente no grupo 1. Não houve diferença significativa entre os sexos para todas as variáveis estudadas. Os hipertensos eram em sua maioria (62%) não controlados e portadores de síndrome metabólica (77%). O exercício físico apresentou efeito benéfico na reclassificação dos hipertensos leves e moderados para a classe de limítrofes sem normalizar a pressão arterial. Não houve efeito do exercício na composição corporal e na homocisteinemia. A suplementação de ácido fólico reduziu a hiperhomocisteinemia em 11% no grupo 1 e 19% no grupo 2 frente a aumentos similares na folacemia. Em relação à pressão arterial houve normalização da homocisteinemia apenas nos hipertensos com pressão arterial normalizada. Os hipertensos limítrofes e leves apresentaram reclassificação da hiperhomocisteinemia limítrofe para moderada. A presença de diurético potencializou a normalização da pressão pelo exercício físico e indiretamente influenciou, nesses casos, a normalização da homocisteína. Entretanto, a maior redução das concentrações médias de homocisteína ocorreu coma oferta de ácido fólico na ausência de diurético. Desta forma, recomenda-se a adequação dietética do folato como adjuvante terapêutico da hipertensão arterial associadamente ao exercício físico e/ou medicamentos / The plasma homocysteine is considered to be a risk indicator for cardiovascular diseases and its association with essential hypertension seems to be important to the worsening of this disease. Physical exercises are efficient to reduce blood pressure and the dietary supplements of folic acid are considered the best way to decrease homocysteine. The aim of this study was investigate the effect of acid folic supplementation and physical exercises on plasmatic levels of homocysteine in essential hypertensive medicated individuals. In order to that, 69 hypertensive subjects were studied (22 men and 47 women) divided in two groups: group 1 medicated with diuretic and group 2 medicated without diuretic. Medical, anthropometrical, dietetic, blood pressure, biochemistry and cardio-respiratory capacity evaluations were performed. These individuals received 500 &#181;g/dia of folic acid supplementation in a cross-over design and were submitted to daily supervisioned physical exercise. They were evaluated every two months until the end of the study. The groups were similar for all variables except for the homocysteine which was significantly greater in the group 1. There were no difference for the gender to all variables. The high blood pressure was not controlled (62%) and 77% of the patients showed metabolic syndrome. The physical exercise showed benefit on the reclassification of slight and moderated hypertensives to border-line hypertension, however, it did not normalize the blood pressure. There were no effects on the body composition and on homocysteine values . Folic acid supplementation decreased 11% of hyperhomocysteinemia in the group 1 and 19% in group 2 face to similar increases in folate. Regarding to high blood pressure subjects the homocysteine normalization occurred only in hypertensive subjects with normalized blood pressure. The border and slightly hypertensive individuals had border-line hyperhomocysteine wich were reclassificated to moderated c1ass. The use of diuretic enhanced blood pressure normalization with the physical exercise and indirectly influenced the plasma homocysteine normalization. However the greater decreasing mean homocysteine concentration occurred with folic acid but in the diuretic absence. So we strong recommend an adequate folate intake as a terapheutical adjuvant to arterial hypertension treatments either with physical exercise and/or antihipertensive drugs.

Rationnel : L’implication établie des progéniteurs endothéliaux circulants dans les phénomènes de néovascularisation chez l’adulte a stimulé la recherche de thérapeutiques angiogènes basées sur la greffe de ces cellules. Deux types cellulaires au phénotype endothélial sont actuellement définis entre autres par leur cinétique d’apparition en culture : les progéniteurs précoces (CFU-EC ou CAC) et tardifs (ECFC). Notre équipe a montré que l’injection thérapeutique de cellules mononucléées de moelle osseuse (BM-MNC) permettait la néovascularisation du site ischémié chez des patients atteints d’artériopathie des membres inférieurs, et que les néovaisseaux formés avaient le phénotype d’ECFC. Nous avons dans un premier temps mesuré les concentrations de différentes protéines modulant l’angiogenèse, chez des patients atteints de pathologies ischémiques et cardiovasculaires, ou impliquant des anomalies vasculaires associées à la fibrose. Ainsi, le transforming growth factor - β1 (TGF-β1) dans la fibrose pulmonaire idiopathique (FPI), la thrombospondine-1 (TSP-1) dans l’artériopathie des membres inférieurs (AMI), et le placental growth factor (PlGF) chez les patients atteints de pathologies cardiovasculaires [syndrome coronarien aigu (SCA), ou patients devant subir une chirurgie de la valve ou un pontage coronarien], se sont distingués comme potentiel biomarqueur plasmatique dans ces pathologies, et ont été étudiés dans la biologie des ECFC humaines.Résultats : Le taux plasmatique de TGF-β1 est augmenté chez les patients atteints de FPI par rapport à la population contrôle ; il a un effet pro-angiogène in vivo (vascularisation des implants de Matrigel®) et in vitro (prolifération et migration des ECFC) via les récepteurs ALK-1, ALK-5 et TGF-βRII. Le taux plasmatique de TSP-1 est augmenté chez les patients artéritiques par rapport à la population contrôle. Par ailleurs les néovaisseaux formés de patients artéritiques ayant été traités par injection locale de BM-MNC expriment la TSP-1. Dans les modèles murins de Matrigel®-plugs et d’ischémie du membre inférieur (IMI), la TSP-1 induit une diminution de la vascularisation des implants ainsi qu’une diminution de la revascularisation du membre ischémié. In vitro, la TSP-1 augmente l’adhésion via un mécanisme N-Terminal dépendant, et diminue le potentiel angiogène (prolifération et migration) des ECFC via sa liaison au récepteur CD47, ce qui active la voie de signalisation SDF-1/CXCR4. Le taux plasmatique de PlGF est augmenté chez les patients atteints de SCA par rapport à 2 populations contrôles ; il est également augmenté chez les patients ayant subit une chirurgie cardiaque. Les PlGF-1 et -2 potentialisent la tubulogenèse des ECFC in vitro via la phosphorylation du récepteur VEGFR1. Cet effet est aboli lorsque le VEGFR1 est inhibé par ARN interférence ou par le composé chimique « 4321 ». De plus ce composé « 4321 » inhibe la vascularisation des implants de Matrigel®, ainsi que la revascularisation du membre ischémié dans le modèle d’IMI.Conclusions : Le TGF-β1 joue un rôle dans le remodelage vasculaire de la FPI via les ECFC ; la TSP-1 est un potentiel biomarqueur de l’angiogenèse induite par les ECFC dans l’AMI ; l’inhibition de la voie PlGF/VEGFR1 module la tubulogenèse induite par les ECFC, cellules impliquées dans la formation de nouveaux vaisseaux. Nous avons ainsi mis en évidence 3 protéines modulant l’angiogenèse dans 3 contextes pathologiques différents, caractérisés par un remodelage vasculaire et où les ECFC sont impliquées dans leurs mécanismes physiopathologiques. Ces 3 protéines se présentent donc comme de potentiels biomarqueurs plasmatiques, modulant les propriétés angiogènes des ECFC et pouvant influencer leur efficacité en tant que produit de thérapie cellulaire. Ces protéines jouent un rôle probable dans l’équilibre homéostatique au décours des pathologies concernées. / Rationale: The pro-angiogenic capacities of endothelial progenitor cells are now well established, and their involvement in neovascularization events in adults has stimulated the research in the field of angiogenic therapy based on transplant of these cells. Current data converge towards the notion of two cell types with endothelial phenotype, defined at least by their kinetics of appearance in culture: early endothelial progenitor cells (CFU-EC or CAC) and late (ECFC). Our team has shown that the therapeutic injection of bone marrow mononuclear cells (BM-MNC) led to neovascularization of the ischemic site in patients with critical limb ischemia, and that the new vessels formed bore the phenotype of ECFC. We initially measured the concentrations of different proteins modulating angiogenesis in patients with ischemic and cardiovascular diseases, or involving vascular abnormalities associated with fibrosis. Thus, the transforming growth factor - ß1 (TGF-ß1) in idiopathic pulmonary fibrosis, the thrombospondin-1 (TSP-1) in peripheral artery disease, and the placental growth factor (PlGF) in patients with cardiovascular diseases [acute coronary syndrome (ACS), patients undergoing valve surgery or coronary artery bypass surgery], emerged as potential plasmatic biomarkers in these pathological settings, and have been studied in the biology of human ECFC.Results: TGF-ß1 plasma level is increased in patients with idiopathic pulmonary fibrosis (IPF) compared to the control population; it exerts a pro-angiogenic effect in vivo (vascularization of Matrigel ®-plugs) and in vitro (proliferation and migration of ECFC) via ALK-1, ALK-5 and TGF-ßRII receptors. TSP-1 plasma level is increased in patients with peripheral artery disease (PAD) compared to the control population. In addition, the new vessels formed in PAD patients treated by local injection of BM-MNC express TSP-1. In murine models of Matrigel ®-plugs and hindlimb ischemia, TSP-1 induces a decrease in plugs vascularization and impaired revascularization of ischemic limb. In vitro, TSP-1 increases ECFC adhesion via an N-terminal dependent mechanism and reduces their angiogenic potential (proliferation and migration) via its binding to CD47 receptor, which activates the SDF-1/CXCR4 signaling pathway. PlGF plasma level is increased in ACS patients compared with the control population and stable angina patients and is also increased in patients undergoing cardiac surgery. PlGF-1 and -2 potentiate ECFC tubulogenesis in vitro via phosphorylation of the VEGFR1 receptor. This effect was abolished when the ECFC VEGFR1 is inhibited by RNA interference or by the chemical compound "4321". In addition this compound "4321" inhibits the vascularization of Matrigel ®-plugs, and revascularization of the ischemic limb in the hindlimb ischemia model.Conclusions: TGF-ß1 is involved in the IPF vascular remodeling through ECFC; TSP-1 is a potential biomarker of angiogenesis induced by ECFC in PAD; the inhibition of the PlGF/VEGFR1 pathway modulates ECFC tubulogenesis, cells involved in the formation of new vessels. We thus identified three proteins that modulate angiogenesis in three different pathological settings characterized by a vascular remodeling and where ECFC are involved in their pathophysiology. These three proteins therefore state as potential plasmatic biomarkers, modulating ECFC angiogenic properties and are able to influence their efficacy as a cell therapy product. These plasmatic biomarkers likely play a role in the homeostasis of those pathologies progress.

L’Annexine A5 (AnxA5) est une protéine soluble se liant aux membranes contenant de la phosphatidylsérine (PS) en présence de calcium (Ca2+). Le rôle central joué par l’AnxA5 dans les processus de réparation membranaire a été récemment mis en évidence. L’AnxA5 possède une très forte affinité pour les membranes biologiques contenant de la PS, cependant son mode de liaison aux membranes n’est pas encore élucidé.La première partie de mon travail de thèse a concerné le développement d’une approche originale d’étude de la liaison de l’AnxA5 à des microsphères de silice fonctionnalisées par une bicouche lipidique (µPSiO2@SLB pour supported lipid bilayer), par cytométrie en flux (FCM). Cette approche m’a permis d’étudier la liaison à l’équilibre et en cinétique à très faible concentration en AnxA5, de l’ordre du picomolaire. Cette approche représente une des méthodes les plus sensibles d’étude de liaison à l’équilibre et la première permettant d’accéder aux constantes cinétiques d’interaction pour l’AnxA5. Cette étude m’a également permis de mettre au point une stratégie de dosage indirect de liposomes contenant de la PS avec une sensibilité de l’ordre du nanogramme de lipides par millilitre.La seconde partie de ma thèse a concerné l’étude de microparticules (MP), fragments de membranes cellulaires présents dans les fluides biologiques. Dans le plasma sanguin la majorité des MP sont d’origine plaquettaire et exposent de la PS. Il existe une corrélation entre la concentration en MP plasmatiques exposant de la PS et le développement de pathologies thrombotiques. La FCM est la méthode de référence dans l’étude des MP cependant leur détection est rendue difficile par leur petite taille. J’ai appliqué aux MP plasmatiques le test de dosage développé pour les liposomes. Les résultats obtenus sont prometteurs et permettent d’envisager le développement d’un test de dosage de l’ensemble des MP exposant de la PS. / Annexin-A5 (AnxA5) is a soluble membrane binding protein that binds to phosphatidylserine (PS) containing membranes in a calcium dependent manner and plays a central role in cell membrane repair processes. AnxA5 has a remarkably high affinity for PS containing membranes, but its binding mechanism remains unclear.The first part of my PhD work was to develop a new method for studying AnxA5 binding using supported lipid bilayer functionalized silica microspheres (µPSiO2@SLB) and Flow Cytometry (FCM). This approach allowed me to describe in details both equilibrium and kinetics of AnxA5 binding at picomolar concentrations in AnxA5. This study is one of the most sensitive for equilibrium binding studies and the first allowing to measure binding kinetics constants for AnxA5. This study also led to the development of a new strategy for determination of liposome concentration with sensitivity in the range of one nanogram of lipid per milliliter. The second part of my work focused on microparticles (MP) that are cell membrane fragments found in biological fluids. In plasma, the vast majority of MP originates from platelets and expresses PS at their surface. There is a correlation between MP concentration in plasma and thrombotic risk. FCM is the “golden standard” of hæmatologic analysis but the majority of MPs are too small to be detected. I have applied the test developed with liposomes for the quantification of MP. The results are promising and allow foreseeing the development of a test able to give the absolute quantity of PS exposing MPs in plasma samples.