Les prairies subalpines abritent de grandes populations d’herbivores sauvages et sont souvent soumises au pâturage domestique. Le but de cette thèse est d’analyser la réponse métabolique des plantes subalpines à l’herbivorie, et d’identifier les conséquences fonctionnelles sur l’écosystème via la décomposabilité des litières. Pour cela, nous avons établi un dispositif d’exclusion des herbivores sur l’alpage d’Armenaz dans le massif des Bauges, dans deux contextes de pâturage différents : extensif par des herbivores sauvages (chamois, Rupicapra rupicapra) et intensif par des herbivores domestiques (génisses, Bos taurus). Nous avons caractérisé une vingtaine d’espèces végétales par des traits fonctionnels foliaires reflétant leur métabolisme de croissance et de défense, en présence ou en absence d’herbivores, et avons estimé la décomposabilité des litières de certaines d’entre elles. Contrairement à l’hypothèse du compromis entre croissance et défense, nos résultats montrent que la présence de composés de défense (polyphénols et lignine) chez les plantes n’est pas reliée à leur stratégie d’acquisition des ressources. Dans notre étude, la réponse des plantes à l’exclusion des herbivores dépend essentiellement : (1) du contexte d’herbivorie, la réponse étant d’avantage marquée dans le contexte du pâturage intensif par les génisses qu’extensif par les chamois, (2) des espèces végétales, suggérant l’existence de différentes stratégies de défense, (3) des conditions environnementales, pouvant être le reflet d’effets d’interaction avec le traitement d’herbivorie. Parmi les réponses observées, l’exclusion des herbivores entraîne chez certaines espèces une diminution de la teneur en composés phénoliques, et plus particulièrement en flavonoïdes, pouvant refléter la relaxation de la synthèse de ces composés de résistance en absence d’herbivores. L’exclusion des chamois n’influence pas la composition chimique des litières ni leur décomposabilité. Cette dernière, parmi les composés étudiés, dépend majoritairement de sa composition en polyphénols. L’identification d’effets contrastés de différents contextes d’herbivorie sur la réponse des plantes et des fonctions écosystémiques qui en découlent nous permet d’améliorer notre compréhension du fonctionnement des prairies subalpines. Ce sont autant d’éléments pour nous aider à mieux appréhender les effets sur l’environnement d’éventuels changements démographiques des populations de grands herbivores. / Alpine grasslands are home to large populations of wild herbivores and are often exposed to additional grazing by domestic herbivores. This study aims to investigate the metabolic response of subalpine plant species to herbivory and to identify the ecosystem functional consequences via the alteration of litter decomposability. For this purpose, we set up a two-year experimental design using exclosures on the Armenaz subalpine grassland from the Bauges massif, under two mammalian herbivory regimes: intensive grazing by domestic heifers (Bos taurus) and extensive grazing by wild chamois (Rupicapra rupicapra). We measured leaf functional traits representative of growth and defense metabolisms on about twenty plant species grown in presence or absence of herbivores, and estimated litter decomposability of some of these species. Contrary to the hypothesis of a growth/defense tradeoff, our results show that the allocation of defensive compounds (polyphenols and lignin) in plants is not related to their nutrient acquisition strategy. In our study, plant responses to grazing exclusion vary according to: (1) herbivory context, plant response being more pronounced in the case of intensive grazing by heifers than extensive grazing by chamois, (2) plant species, suggesting that they rely on different defensive strategies, (3) environmental conditions, which may reflect the interactive effects with grazing treatment. One of the observed responses of plants to grazing exclusion is the decrease of their phenolic content, and more specifically flavonoids, which can be explained by the relaxation of the production of plant defenses when herbivores are no longer present. Chamois exclusion has no influence on chemical composition and decomposability of litters, the latter being best explained by its phenolic content. Revealing the contrasted effects of wild and domestic mammalian herbivores on vegetation and related soil processes allow us to improve our understanding of subalpine grasslands functioning. This should help us to better predict the effects of potential demographic variations of herbivore populations on environment.

Este trabalho teve como objetivo comparar diferenças entre as progênies de touros representativos raça Nelore e de três grupos genéticos, Nelore, Angus x Nelore e Brahman x Nelore, quanto ao desempenho pós desmame e quanto às características de carcaça medidas in vivo por ultra-som. Foram selecionados 14 touros representativos da raça Nelore, dois da raça Aberdeen Angus e um touro Brahman. Estes touros foram acasalados com 400 vacas Nelore, que deram origem a uma progênie de 158 machos e 148 fêmeas. Os animais foram criados e recriados a pasto até os 19 meses de idade e, então, terminados em confinamento recebendo dieta com 14,2% de PB e 71,2% de NDT, numa relação volumoso:concentrado de 23:77. Durante a fase de recria foram realizadas três pesagens e uma medida de carcaça por ultra-sonografia, aos 483 dias de idade em média. A fase de terminação em confinamento compreendeu cinco pesagens, realizadas aproximadamente a cada 28 dias, e três medidas de carcaça por ultra-sonografia, sendo que os animais apresentavam as seguintes idades médias em cada avaliação: 570, 598 e 677 dias, respectivamente. As medidas tomadas foram: área de olho de lombo (AOL, no músculo Longissimus dorsi) e espessura de gordura subcutânea entre a 12ª e 13ª costela (EG) e na garupa (P8, sobre o músculo Biceps femoris). Os animais permaneceram confinados até atingirem aproximadamente 5 mm de espessura de gordura subcutânea medida entre as 12ª e 13ª costelas no músculo Longissimus dorsi, no caso, por 127 dias, entre os meses de julho e novembro de 2004. Os animais foram abatidos com 24 a 26 meses de idade. Após o abate, foram coletados medidas de peso da carcaça quente (PCQ) e rendimento de carcaça. O grupo genético com melhor desempenho pós-desmama foi o grupo Angus x Nelore, seguido de Brahman x Nelore e por fim os animais Nelore puros, sendo que os machos foram superiores às fêmeas. Entretanto, durante o período de terminação em confinamento, as fêmeas apresentaram ganho de peso semelhante aos machos castrados, porém tiveram taxas de ganho de músculo e gordura superiores, em conseqüência da sua maior taxa de maturação. Foram verificados maiores ganhos em AOL nos Angus x Nelore, mostrando a aptidão para produção de carne da raça taurina. As diferenças em ganhos de gordura foram pequenas, provavelmente devido à qualidade da dieta que não permitiu que todo o potencial genético para terminação fosse expressado. Quando o estudo se baseou somente na raça Nelore, foram verificadas diferenças nas características de crescimento pós-desmama e durante a terminação em confinamento, entre as progênies dos touros da raça Nelore, indicativas da existência da variabilidade genética, o que permitiria esperar uma resposta à seleção de animais para as mesmas. Entretanto não houve efeito de touro para velocidade de acabamento e crescimento muscular entre as progênies estudadas. / This work aimed to evaluate differences among sires selected to represent the Nellore breed and among three breed groups (Nellore, Angus x Nellore, Brahman x Nellore) as to their progenies’ post-weaning performance and carcass traits, measured by real time ultrasound. A total of 14, two and one sires of the Nellore, Angus and Brahman breeds, respectively, were mated to 400 Nellore cows, producing a progeny of 158 steers and 148 heifers. The animals were raised on pasture until 19 months of age, then finished in feedlot on a diet containing 14.2% crude protein and 71.2% TDN, with a roughage:concentrate ratio of 23:77. During the growing phase on pasture, the animals were weighed three times and scanned at 483 days of age for Longissimus muscle area (LMA), back fat thickness over the 12th - 13th ribs (backfat) and rump fat (P8 fat). During the finishing phase in feedlot, between July and November 2004, cattle were weighed every 28 days and scanned for LMA, backfat and P8 fat at 570, 598 and 677 days of age. The backfat thickness end-point (5 mm) was reached after 127 days on feed, Animals were slaughtered at 24 to 26 months of age. At slaughter, hot carcass weight and dressing percentage were measured. There were effects of breed group on performance during the growing phase. Angus-sired calves had better performance than Brahman-sire and Nellore-sired calves and Brahman x Nellore calves were superior to straight bred Nellore. Males had better performance than females. However, during the finishing phase, there was no sex effect on weight gain and females had greater muscle and fat gain rate than males, perhaps due to their greater maturity rate. Angus x Nellore animals had greater LMA, showing the meat producing ability of Bos taurus breeds. Breed differences as to backfat gains were slight. This was likely due to diet quality which didn’t allow the animals’ genetic potential to be fully expressed. Within the Nellore breed, post-weaning growth traits were different among the sires, indicating existence of genetic variability. Nevertheless, there were no effects of sire on backfat gain and muscle growth among the progenies studied.

Diversos estudos indicam que touros europeus (Bos taurus) apresentam uma maior susceptibilidade ao estresse térmico com um consequentemente aumento do estresse oxidativo. Este, por sua vez, leva a uma diminuição na qualidade espermática. O espermatozoide possui, fisiologicamente, uma grande quantidade de ácidos graxos poli-insaturados (PUFAs) em sua membrana. As ligações duplas entre carbonos desses PUFAs confere a membrana plasmática uma maior fluidez, necessária aos processos fisiológicos do espermatozoide como motilidade e fertilização. No entanto essas ligações são mais facilmente oxidadas e, portanto, mais susceptíveis a peroxidação lipídica. Sendo assim, a suplementação com PUFAs promoveria uma melhoria na qualidade espermática, porém, tornaria o espermatozoide ainda mais susceptível à peroxidação lipídica. A vitamina E é um antioxidante chave na prevenção do estresse oxidativo através da interceptação do radical hidroxila, protegendo os PUFAs da membrana espermática contra a peroxidação lipídica. Portanto, uma alternativa para melhorar a qualidade espermática de touros europeus submetidos ao estresse térmico, seria a suplementação com PUFAs associada a um tratamento com vitamina E para prevenir um possível efeito deletério oxidativo desses ácidos graxos. Estudos sugerem uma importante relação entre os testes convencionais de avaliação de sêmen e fertilidade. No entanto, a limitação destes testes também tem sido reconhecida, principalmente em casos de infertilidade idiopática. Estudos demonstram que os testes funcionais de avaliação do sêmen, podem ser uma alternativa importante para a avaliação da capacidade fecundante de uma determinada amostra seminal. A interação entre espermatozóide e zona pelúcida ocorre através da ligação de receptores acrossomais às proteínas ZP3 e ZP2 do oócito. A membrana perivitelínica de ovos de galinha possui uma homologia com essas proteínas, permitindo sua ligação com espermatozóides de diversas espécies. Sendo assim, o teste de ligação espermática a essa membrana parece ser bastante eficiente para avaliar a capacidade fecundante do sêmen. No presente estudo, 16 touros Bos taurus foram submetidos à insulação testicular durante 4 dias e submetidos a um arranjo fatorial 2x2, sendo os fatores: dieta rica em PUFAs (Megalac®, 4 Kg ao dia) e tratamento com Vitamina E (Monovin E®, 3000 UI a cada 13 dias) durante 60 dias. As amostras seminais criopreservadas/descongeladas foram submetidas à avaliação da análise computadorizada da motilidade (CASA; Hamilton Thorne, Ivos 12.3, USA), integridade de membrana (Eosina Nigrosina), integridade acrossomal (Fast Green Rosa Bengala), atividade mitocondrial (33 diaminobenzidina), susceptibilidade ao estresse oxidativo (Substâncias reativas ao ácido tiobarbitúrico - TBARS), integridade de DNA (SCSA) e o teste de ligação de espermatozóides à membrana vitelínica de ovos de galinha, que foi previamente padronizado. O tratamento com vitamina E apresentou efeitos benéficos para algumas características espermáticas, sendo o contrário observado pelo tratamento com PUFAS, quando ambos foram analisados isoladamente. No entanto, a associação entre estes tratamentos não foi eficiente para melhorar a qualidade espermática assim como a capacidade fecundante avaliada através do teste de ligação espermática em membrana perivitelínica de ovos de galinha. / Several studies suggest a high susceptibility of European bulls (Bos taurus) to the heat stress, which will lead to an increase on oxidative stress and consequent impairment on sperm viability. Sperm membranes are rich in poly-unsaturated fatty acids (PUFAs). The carbon-carbon double bounds (insaturations) that characterize these PUFAs confer the necessary fluidity to the sperm membrane, essential to motility and fecundation capacity. However, these insaturations are more unstable and, therefore, more susceptible to the lipid peroxidation (i.e., attack of reactive oxygen species to the lipids). Hence, PUFA supplementation would improve certain sperm characteristics but increase the already high susceptibility to the oxidative stress. Vitamin E is a key antioxidante on oxidative stress prevention by intercepting hydroxyl radical and protecting sperm membrane PUFAS against lipid peroxidation. Therefore, an alternative to improve sperm quality in heat stressed european bulls would be a PUFA supplementation associated to a Vitamin E therapy in order to avoid a possible deleterious oxidative effects of these fatty acids. Estudies suggest an important relationship between conventional tests for sperm evaluation and fertility. However, limitations have been also verified, especially in cases of idiopathic infertility. Thus, tests to evaluate sperm functionality may be an interesting alternative to assess fecundation capacity of semen samples. The binding of sperm to the zona pellucida occurs initially by the interaction between acrosomal receptors and oocyte proteins ZP3 and ZP2. The perivitelline membrane of chicken eggs present homologies to zona pellucida proteins, which allows sperm from several species to bind to this membrane. Thus, the sperm binding test to the perivitelline membrane may be an efficient method to asses the fecundation capacity in semen of bulls. In the present study, 16 Bos taurus bulls were submitted to testicular insulation for 4 day. Animals were then submitted to a 2x2 factorial design, with the factor represented by a PUFA oral supplementation (Megalac®, 4 Kg for day) and the treatment with Vitamin E (Monovin E®, 3000 UI, each 13 days) for 60 days. Samples were cryopreserved/thawed and analyzed for motility variables by the computer assisted sperm analysis (CASA; Hamilton Thorne, Ivos 12.3, USA), membrane integrity (Eosin / Nigrosin), acrosome integrity (Fast Green / Bengal Rose), mitochondrial activity (33 diaminobenzidine), susceptibility to the oxidative stress (tiobarbituric acid reactive substances - TBARS), DNA integrity (SCSA) and the sperm binding test to the chicken egg perivitelline membrane, which was previously validated. Results demonstrated a beneficial effect of Vitamin E on some sperm characteristics, with the inverse effect observed for PUFA when both treatments were considered individually. However, the association between both treatments were not efficient to improve sperm quality as well as the fecundation capacity as evaluated by the sperm binding test to the chicken perivitelline membrane.

Orientador: Humberto Tonhati / Coorientador: Luciana Correia de Almeida Regitano / Coorientador: Lúcia Galvão de Albuquerque / Banca: José Fernando Garcia / Banca: Simone Cristina Méo Niciura / Resumo: Na bovinocultura de corte, a probabilidade de prenhez precoce é uma característica de grande relevância devido ao alto valor econômico associado. Essa característica apresenta elevados valores de herdabilidade e há possibilidade de ganho genético por seleção. Todavia, a característica é de limiar com distribuição binária e suposições são feitas no intuito de analisá-la. Marcadores moleculares podem vir a auxiliar na avaliação genética desses animais, além de promover diminuição do intervalo de gerações. A proteína JY-1 é uma proteína específica dos oócitos de bovinos e que atua nas células da granulosa em co-ação com o LH e no desenvolvimento embrionário inicial podendo influenciar a chance de prenhez. Marcadores moleculares foram usados para estudar o gene JY-1 e foram encontrados sete polimorfismos do tipo SNP no éxon 3. As posições dos SNPs no referido éxon e as substituições são: 163 (T/C), 281(T/C), 321(T/C), 532(T/C), 652 (A/G), 679(T/C) e 722 (G/C) (Genbank: JN592587 JF262042.2). O SNP 163 está em região codificante e causa substituição de uma prolina por uma leucina. Os demais SNPs estão em região 3'UTR. Os SNPs 163, 281, 321 e 679 foram genotipados em 297 novilhas da raça Nelore. Os SNPs 163, 321 e 679 estavam em desequilíbrio de ligação entre si e em equilíbrio com o 281. Os genótipos não apresentaram correlação com probabilidade de prenhez precoce e características de crescimento (P<0,05) / Abstract: In beef cattle production, the sexual precocity is important because of the high associated economic value. The heritability values are high which permits genetic gain by selection. However, it is a threshold trait with binary distribution and suppositions are done in order to analyze it. So, molecular markers may auxiliary the genetic evaluation of these animals and reduce the generation interval. The JY-1 protein is an oocyte specific cattle protein and acts in granulose cells with LH and also in early embryo development and may influence sexual precocity. The gene of the protein was analyzed by molecular markers and seven SNP polymorphisms were found in exon 3. The SNPs positions in the exon and the substitutions were: 163 (T/C), 281(T/C), 321(T/C), 532(T/C), 652 (A/G), 679(T/C) e 722 (G/C) (Genbank: JN592587 JF262042.2) The SNP 163 is in CDS region and it causes a substitution of a prolin by a leucin. The others SNPs are in 3'UTR region. The SNPs 163, 281, 321 and 679 were genotyped in 297 Nellore heifers. The SNPs 163, 321 and 679 were in linkage disequilibrium among them and the SNP 281 was in equilibrium. The genotypes did not show correlation with sexual precocity and growth traits (P<0.05) / Mestre

Orientador: Humberto Tonhati / Coorientador: Fábio Ricardo Pablos de Souza / Coorientador: Lúcia Galvão de Albuquerque / Banca: Janete Apparecida Desidério / Banca: Maria Eugênia Zerlloti Mercadante / Resumo: Na pecuária de corte, características de crescimento e de carcaça são de grande importância para programas de melhoramento genético. Para acelerar o processo de seleção, os marcadores moleculares auxiliam o estudo dessas características que são influenciadas por efeitos poligênicos. Os genes ACRP30, PPARGC1 e OLR1, foram associados à características ligadas a produção em Bos taurus taurus. Esses genes estão ligados aos processos de sínteses de lipídios, síntese metabólica e acúmulo de gordura no tecido adiposo. Em nosso estudo, foram genotipados 639 animais, através da técnica de PCR-RFLP, para os genes ACRP30/BsrI, PPARGC1/NheI e OLR/PstI, onde foram detectados polimorfismos correspondentes aos sítios de restrição. Para as linhas de seleção Controle (NeC), Seleção(NeS) e Tradição(NeT) em bovinos Nelore, do Instituto de Zootecnia de Sertãozinho-IZ, foram feitas associações com características de peso ao nascer, peso no desmame, peso ao sobreano das fêmeas, altura ao sobreano da garupa das fêmeas, peso ao sobreano dos machos, altura ao sobreano da garupa dos machos, área de olho de lombo, espessura de gordura da garupa, espessura de gordura subcutânea e polimorfismos nos genes estudados, utilizado o PROC MIXED, do programa SAS/STAT 9.1.3. para análises de associações e Teste de Fisher para diferenciação entre populações do programa Genepop. A correção dos testes de hipóteses para comparações múltiplas foi feita pelo método de Bonferroni Os genótipos apresentados foram associados com as características de crescimento e carcaça, não apresentando efeitos significativos na população estudada / Abstract: In beef cattle production, growth and carcass traits are of great importance for animal breeding programs. To speed up the selection process, molecular markers help to study traits that are influenced by polygenic effects. The ACRP30, PPARGC1 and OLR1 genes were validated in some studies for traits related to production in Bos taurus. This genes are linked to processes of lipids and metabolic synthesis and accumulation of fat in the adipose tissue. In this study, 639 animals were genotyped by PCR-RFLP, for genes ACRP30/BsrI, PPARGC1/NheI and OLR/PstI, where polymorphisms were detected, corresponding to the restriction sites. For selection lines Control (NeC), Selection (NeS) and Tradition (NeT) in Nellore cattle, from the Institute of Zootecnia from Sertãozinho - IZ, associations were made with polymorphisms in the studied genes and the trais birth weight, weaning weight, yearling weight, yearling rump height, ribeye area, rump fat thickness and subcutaneous fat thickness, using the PROC MIXED from SAS/STAT 9.1.3. program for association analysis, and Fisher Test for population differentiation from Genepop program. The correctness of the hypothesis tests for multiple comparisons was made using the Bonferroni. The genotypes found were associated with the with growth and carcass traits but showed no significant effects on the studied population / Mestre

A espécie Bos indicus, particularmente a raça Nelore, é grande maioria no rebanho bovino da região acima do trópico no Brasil. Embora a habilidade desses animais em resistir às doenças parasitárias, condições climáticas e pastagens de baixa qualidade enalteçam a utilização em larga escala desta raça, estes animais não são considerados bons conversores de alimento e, conseqüentemente, precoces em comparação aos seus homólogos Bos taurus. Durante a formação das raças zebuínas brasileiras, houve uma participação das linhas maternas de Bos taurus, que pode ser demonstrada pela contribuição majoritária do genoma mitocondrial desta subespécie. Embora em escala muito menor, estima-se que exista também uma participação destas linhas maternas no genoma nuclear. O objetivo deste trabalho foi iniciar os estudos para estimar esta participação. Para os estudos foram utilizados 104 animais da raça Nelore e 8 animais de diferentes raças européias. Cinco regiões do DNA que produzem fragmentos microssatélites taurus/indicus específicos (HEL1, HEL9, ETH225, ILSTS005 e INRA063) foram amplificadas com a utilização de primers marcados com sondas fluorescentes. Os fragmentos foram submetidos à eletroforese em gel de poliacrilamida desnaturante 6% e visualizados após excitação com laser. No total foram encontrados 23 alelos para os microssatélites analisados o que representa uma média de 4,6±1,82 alelos por locus. Amplificou-se também uma região do DNA satélite 1711b que posteriormente foi digerida com a enzima de restrição Msp I. Verificou-se a existência de três possíveis genótipos entre os animais Nelore mtDNA Bos taurus e mtDNA Bos indicus. Os animais europeus analisados apresentaram sempre o mesmo padrão de restrição. A comparação dos componentes de variância do tamanho dos alelos intra e inter população usando os fragmentos microssatélites permitem a separação dos animais Bos taurus dos animais Nelore, mas não dos Nelore de origem materna distinta. No entanto, a freqüência de alelos indicus específicos nos microssatélites e de padrões de digestão do DNA satélite também indicus específicos sugerem uma participação da ordem de aproximadamente 6% do genoma taurus na população de gado Nelore. / Bos indicus specie, especially Nellore breed is responsible for the majority of Brazilian tropical herd. These animals are notably capable to endure parasite infection as well as hot weather and low quality feed. In one hand this qualities suggest the large scale application of this breed, but in other hand this same breed is well characterized as bad food converter and consequently far from having good precocity status compared with its Bos taurus homologues. It has been reported a matrilineal European participation in Zebu cattle since its introduction in American lands. This hybridization is confirmed by the majority contribution of Bos taurus mtDNA in these animals. Although in a much lower frequency, we hypothesize a Bos taurus cow participation in nuclei genome. The main aim of this work was to give the firsts steps towards the estimation of this participation. A total of 104 Nellore and 8 animals of different European breeds were used for DNA analysis. Five microsatellites fragments (HEL1, HEL9, ETH225, ILSTS005 e INRA063) were amplified applying primers with fluorescent dye. Amplified fragments were used in 6% polyacrilamide electrophoresis and visualized after laser excitation. Overall 23 alleles were detected averaging 4.6±1,82/locus. Variance components of microsatellites allele size comparisons allowed the formation of two clusters separating both subspecies. No significant variation was observed between Nellore with different maternal origins. A satellite 1711b DNA was also amplified and digested with the restriction enzyme Msp I. Three possible genotypes were identified in Nellore animals harboring B. taurus and B. indicus mtDNA. European originated animals always showed the same restriction pattern. Finally B. indicus specific microsatellite allele and satellite 1711b digestion patterns frequency allowed the estimation of 6% of B. taurus contribution in purebred Nellore. These results are discussed in terms of application in cattle genetic improvement.

O objetivo deste estudo foi determinar os efeitos da aplicação de hCG no quarto dia após a inseminação a tempo fixo (IATF) sobre o tamanho do corpo lúteo (CL) existente, a indução de CL acessórios, a concentração de progesterona (P4) sérica e taxa de prenhez de vacas de corte em lactação. Nos três experimentos, vacas de corte multíparas (n=569), cruza Bos taurus, com período pós-parto entre 45 e 70 dias, foram sincronizadas com a administração de 2mg de benzoato de estradiol i.m. e a inserção de um dispositivo intravaginal contendo 0,750g de P4 (Dia -9). Sete dias após, administraram-se 150 μg de D-cloprostenol e 0,5 mg de cipionato de estradiol, i.m., no momento da retirada do dispositivo (Dia -2). Sessenta vacas de corte em lactação foram divididas aleatoriamente em dois grupos: hCG (n=30) tratadas com 1500UI i.m. de hCG e Controle (n=30) injetadas com 1,5 mL de solução salina i.m. no dia 4 depois do estro (Dia 0). Realizaram-se coletas sanguíneas dos animais para mensuração da concentração sérica de P4 nos Dias 4, 7, 10 e 14 do ciclo estral. Para acompanhamento da dinâmica ovariana, sessenta animais foram divididos aleatoriamente nos grupos hCG (n=30) ou Controle (n=30) e receberam o mesmo protocolo hormonal como citado anteriormente. No Dia 0, os ovários foram examinados por ultrassonografia transretal para determinar o diâmetro do folículo ovulatório; no Dia 4, o diâmetro do FD e presença do CL e no Dia 7 para mensurar a área luteal do CL e a presença de CL acessório. Quinhentas e sessenta e nove vacas (hCG, n= 269 e Controle, n= 300) foram inseminadas a tempo fixo 52-56 horas depois da retirada dos implantes de P4 e tratadas no Dia 4, conforme o protocolo descrito. O diagnóstico de prenhez foi realizado por ultrassonografia 30 dias após a IATF. Os resultados observados, mostraram que vacas tratadas com hCG no Dia 4 apresentaram maiores concentrações sérica de P4 no Dia 7, comparadas com as do grupo Controle (4,45 vs. 3,37ng/mL, respectivamente; p<0,05), mas níveis semelhantes de P4 nos Dias 10 e 14. Os animais do grupo hCG apresentaram CL com maior (p<0,01) área luteal no Dia 7, em relação ao grupo Controle (3,52 cm² vs. 2,66 cm², respectivamente) e uma incidência de 29,6 % de indução da ovulação do FD presente no Dia 4. Observou-se uma taxa de prenhez maior (p=0,071) no grupo tratado com hCG (53,9%) comparando-se com as vacas não tratadas (46,3%). Conclui-se que a administração de hCG no quarto dia do ciclo estral promove o aumento da área do CL, melhora função luteal, pode induzir a formação de CL acessório e tende a aumentar a taxa de prenhez de vacas de corte em lactação. / The aim of this study was to determine the effects of injection of hCG on the fourth day after the TAI on the size of the existing corpus luteum (CL), the induction of CL accessories, the concentration of progesterone (P4) levels and the possibility of increasing the rate of pregnancy of lactating beef cows subjected to synchronization of estrus and ovulation. Multiparous beef cows (n= 569), Bos taurus crossbreed with postpartum between 45 and 70 days were synchronized by administration of 2mg of estradiol benzoate im and the insertion of an intravaginal device containing 0,750g of P4 (Day -9). Seven days later, were administered 150mg of D- cloprostenol 0.5 mg of estradiol cypionate, im, at the time of device removal (Day -2). Sixty lactating beef cows were randomly divided into two groups: hCG (n= 30) treated with im 1500UI hCG and Control (n= 30) injected with 1.5 ml of saline im on day 4 after estrus (Day 0). There were blood collections of animals for measurement by radioimmunoassay of serum P4 on Days 4, 7, 10 and 14 of the estrous cycle. To evaluate the ovarian dinamics, sixty animals (hCG, n= 30 and Control, n = 30) received the same hormonal protocol and were uniformly distributed into groups according to the diameter of the ovulatory follicles on day 0 (estrus). On Day 0, the ovaries were examined by transrectal ultrasonography to determine the diameter of the ovulatory follicle, on Day 4, the diameter of the dominant follicle (DF) and the presence of CL on Day 7 to measure the area of the luteal CL and the presence of accessory CL. Five hundred and sixty-nine cows (hCG, n= 269 and Control, n= 300) were inseminated at a fixed time 52 to 56 hours after removal of the P4 devices treated on Day 4, according to the protocol described previously. The diagnosis of pregnancy were performed by ultrasonography 30 days after TAI. The results demonstrate that cows treated with hCG on Day 4 showed higher serum concentrations of P4 on Day 7, compared with the control group (4.45 vs. 3.37ng/mL, respectively, (p<0.05), but similar levels of P4 on Days 10 and 14. The animals of hCG group had greater (p<0,01) luteal area on Day 7, compared to the Control group (3.52cm² vs. 2.66cm², respectively) and a 29.6% incidence of induction DF ovulation present on Day 4. We observed a higher rate of pregnancy (p= 0.071) in the group treated with hCG (53.9%) comparing with untreated cows (46.3%). We conclude that administration of hCG on the fourth day of the estrous cycle increase the CL area, improves luteal function, can induce the formation of accessory CL and tends to increase the pregnancy rate of lactating beef cows.

All contemporary taurine cattle originated some 10,000 years ago when their wild ancestor, the aurochs, was domesticated in the Near East. Although the aurochs was widespread also in Europe, there is no evidence for a local domestication. The aurochs has been extinct since 1627 and therefore little is known about its biology. Following domestication, cattle were selected for traits of interest to humans. All modern cattle breeds were developed in the 19th century and the only sources of information about prehistoric breeding practices, and breeds, come from a few ancient Roman Empire and medieval European written accounts. The aim for this thesis was to investigate the effects early selection may have had on the cattle genome and to investigate genetic variation in European aurochs. Using second-generation sequencing and coalescent simulation analyses of aurochs Y chromosomal DNA, I estimated effective population size to between 20,000-80,000 aurochs bulls, indicating that a large population was present when domestic cattle entered Europe. A Y chromosomal SNP revealed that the two male lineages present in modern cattle were also present in European aurochs, and that the frequency of these lineages in domestic cattle fluctuated over time. This indicates that cattle were mobile and that bottlenecks, possibly due to selective breeding, occurred. I used nuclear SNPs to trace genetic variation in North European cattle through time and show that when genetics is combined with archaeology and osteology, even small but notable changes in the use of cattle can be detected. There has been a significant decrease in genetic variation over time, with the most dramatic changes associated with the formation of breeds during the 19th century.

The cattle tick, Rhipicephalus Boophilus microplus (formerly Boophilus microplus), is the most significant parasite of cattle in Australia and threatens the improvement of cattle production in tropical and subtropical countries worldwide. Some cattle breeds of mainly Bos indicus origin develop a strong resistance to infestation with R. B. microplus, while other breeds of mainly Bos taurus origin will succumb to anaemia and ‘tick worry’ in tick-infested pastures. Resistance to cattle tick infestation is primarily manifest against the larval stage and results in the immature tick failing to make a successful attachment and obtain a meal. It is widely accepted that resistance to tick infestation in cattle is immunologically mediated and involves both cell-mediated and humoral immune responses. Previous research has demonstrated a role for the hypersensitivity response in the rejection of larvae from resistant B. taurus cattle. The immune response to R. B. microplus infestation in B. indicus and B. indicus × B. taurus cattle has not been studied in great detail and this thesis aimed to describe those responses of highly tick-resistant cattle that differ from those of tick-susceptible cattle. A pilot trial was conducted using six tick-resistant Brahman heifers (B. indicus) and six tick-susceptible Holstein-Friesian heifers (B. taurus). The animals were artificially infested for several weeks and blood samples obtained weekly for three weeks during the height of infestation, and on one occasion, skin biopsies obtained. It was found that significant differences existed between the two breeds with respect to the percentage of cellular subsets comprising the peripheral blood mononuclear cell (PBMC) population, cytokine expression by peripheral blood leukocytes (PBL), and levels of tick-specific immunoglobulin G1 (IgG1) antibodies measured in the peripheral circulation. These results in combination with whole genome expression analysis of circulating PBL suggested that the B. indicus cattle had developed a stabilised T cell-mediated response to tick infestation evidenced by their cellular profile and leukocyte cytokine spectrum. The B. taurus cattle demonstrated cellular and gene expression profiles consistent with a sustained innate, inflammatory response to infestation, although high tick-specific IgG1 levels suggested that these animals had also developed a T cell response to infestation. Gene expression analyses using quantitative PCR (qPCR) and whole genome expression platforms indicated that the B. taurus cattle elicited an innate inflammatory response in the skin at the site of larval attachment. This was supported by histological examination of the tick-attachment sites. Conversely, the B. indicus cattle appeared to limit the inflammatory response and instead exhibited increased expression of genes involved in wound healing and the structural integrity of the skin. A second trial was conducted using a group of tick-naïve Santa-Gertrudis heifers to eliminate any innate breed differences that may have confounded the results obtained in the pilot trial. Thirty-five animals were obtained from a tick-free region of Australia; thirty animals were artificially infested weekly for thirteen weeks while five animals remained at a tick-free quarantine property to serve as a control group. Following thirteen weeks of tick infestation the animals in this trial exhibited divergent tick-resistance phenotypes and those carrying the lowest and highest numbers of ticks were classified into tick-resistant (n = 6 animals) and tick-susceptible (n = 6 animals) groups, respectively. The larger number of animals in this study and repeated measurements over the course of the trial enabled a detailed analysis of the effect of tick infestation across all animals, and also within the groups identified as highly resistant or highly susceptible. A cell-mediated response to tick-infestation was detected in all tick-infested animals by twenty-one days after the initial infestation, a response which waned towards the end of the trial when the antibody response became more dominant. Similar to the pilot trial, the tick-susceptible animals developed significantly higher levels of tick-specific IgG1 antibodies compared with the tick-resistant group. Some differences were observed between tick-resistant and tick-susceptible animals with respect to the percentage of cellular subsets comprising the PBMC population. Skin biopsies were taken from these animals prior to initial infestation and then at twenty-one days and ninety-one days after the initial infestation and whole genome expression profiles were produced and analysed. At twenty-one days post infestation both tick-resistant and tick-susceptible animals demonstrated an upregulation of genes involved in leukocyte migration and complement activation, suggestive of an inflammatory response. While this response persisted in the tick-susceptible group at ninety-one days post-infestation, it appeared to lessen in the tick-resistant group. A dominant interleukin-8 (IL-8) response was observed in the skin of the tick-susceptible animals at both time points post-infestation that was not observed in the tick-resistant animals. This thesis demonstrates that tick-susceptible animals respond to infestation with R. B. microplus with an apparently non-protective antibody response, and that tick-resistant animals demonstrate different structural responses in the skin at the tick-attachment site compared with the intense inflammatory response observed in tick-susceptible animals. The results presented here suggest that the extreme susceptibility of pure B. taurus and some B. taurus × B. indicus individuals may be due to their heightened ability to recognise and respond to tick antigens introduced into the host via tick saliva during the blood feeding process. These animals’ increased ability to recognise and respond to these proteins may act to set up a chronic state of inflammation that is beneficial to the tick through increased capillary permeability providing a suitable environment for tick feeding and survival.

Submitted by Nathália Faria da Silva (nathaliafsilva.ufv@gmail.com) on 2017-06-07T14:44:18Z No. of bitstreams: 1 texto completo.pdf: 274417 bytes, checksum: c9c56c059a20f3a90aab2b24c081fdc8 (MD5) / Made available in DSpace on 2017-06-07T14:44:18Z (GMT). No. of bitstreams: 1 texto completo.pdf: 274417 bytes, checksum: c9c56c059a20f3a90aab2b24c081fdc8 (MD5) Previous issue date: 2000-12-07 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O experimento foi conduzido na Fazenda Mundo Novo, município de Brotas - SP, de outubro de 1999 a março de 2000. Os objetivos foram determinar a eficiência do teste da libido realizado em curral, comparando-o ao teste realizado a campo, e relacioná-los à taxa de gestação em estação de monta de 120 dias; testar a tabela de classificação da libido proposta pelo CBRA (1998), para animais da raça Nelore (Bos taurus indicus); propor modificações no teste, e confirmar a relação comportamento sexual/aspectos físicos e morfológicos do sêmen. Foram utilizados 13 touros, todos classificados como aptos à reprodução, avaliados por exame andrológico (exame biométrico dos órgãos sexuais e exame físico e morfológico do sêmen e avaliação clínica), e submetidos ao teste da libido em curral, por 3 h. Destes, 9 touros foram submetidos à avaliação a campo, com proporção touro/vaca 1:33. A cada 21 dias, foi diagnosticada a gestação por ultra-sonografia. Observou-se que, quanto maior o tempo de teste, maiores foram os escores de classificação, sendo que nenhum touro conseguiu efetuar serviço completo dentro dos 10 minutos do teste. Touro classificado como muito bom teve 84,80% de prenhez, porém touros questionáveis tiveram 86,67% e 96,55% ao final da estação. As correlações entre características físicas e morfológicas do sêmen ficaram próximas de zero ou nulas (p>0,05). Os touros reconhecem a fêmea que está em estro pelo ato de cheirar ou lamber o corpo da fêmea, cheirar ou lamber a vulva, seguindo-se ou não o reflexo de Flehmen. A receptividade das fêmeas é testada e os machos efetuam o coito precedido por eventos tais como exposição de pênis, reflexo de monta, tentativa de monta, monta abortada e serviço completo ou cópula. / The experiment was carried out during the period from October 1999 to March 2000 in Fazenda Mundo Novo, Brotas county - SP. The objectives were: to determine the efficiency of the libido test performed in corral, by comparing it to the one accomplished at field, as well as to relate them to the gestation rate over a reproductive period of 120 days; to test the libido classification table proposed by CBRA (1998) for animals of the race Nelore (Bos taurus indicus); to propose modifications for the test and confirm the relationship sexual behavior/ physical and morphologic aspects of the semen. Thirteen bulls were used, all being classified as reproductors and evaluated by breeding soundness evaluation (sexual organ biometric evaluation, and physical and morphologic evaluation of the semen and clinical evaluation), and submitted to the libido test for 3 hours in corral. From these, nine bulls were submitted to the evaluation at field, with a bull/cow proportion of 1:33. At every 21 days, the gestation was diagnosed by ultrasonographic test. It was observed that as longer was the test time as higher were the classification scores, and no bull was able to perform a complete mating service within 10 minutes under testing. The bull classified as xiiivery good showed a conception rate of 84.80%, while the questionable bulls showed 86.67% and 96.55% at the end of the mating period. The correlation among physical and morphologic characteristics of the semen were close to zero or null (p>0.05). The bulls do recognize the female being in oestrous by either the act of smelling or licking the female body or smelling or licking the vulva, following or not the Flehmen reflex. The receptivity of the females is tested and the males perform the coitus preceded by some events such as: penis exhibition, mating reflex, mating attempt, aborted mating) and complete mating service or copulation.