Two studies were conducted to assess factors affecting estrogen excretion in dairy heifers. The objective of the first study was to quantify estrogenic activity in feces and urine during the estrous cycle. Ten non-pregnant Holstein heifers were fed the same diet for 28 d. Plasma, feces, and urine samples were collected daily. Plasma 17-β estradiol (17-β E2) was quantified with RIA and used to confirm day of estrous. Feces and urine samples from days -12, -6, -2, -1, 0, 1, 2, 6, 12 of the estrous cycle were analyzed with RIA and Yeast Estrogen Screen (YES) bioassay. Plasma 17-β E2 concentrations peaked on day of estrus, with feces and urine estrogenic excretion peaking a day after. The objective of the second study was to quantify variation in estrogenic activity in feces and urine due to increased dietary phytoestrogen content. Ten Holstein heifers were randomly assigned to treatment sequence in a two-period crossover design. Dietary treatments consisted of grass or red clover hay, and necessary supplements. Feces and urine samples were collected and pooled for analysis. Estrogenic activities of pooled samples were quantified using the YES bioassay. Estrogenic excretion in feces and urine was higher for heifers fed red clover hay. Fecal and urine samples from five heifers were analyzed using LC/MS/MS to quantify excretion of phytoestrogenic compounds. Heifers fed red clover hay excreted more equol than heifers fed grass. Identifying sources of variation in estrogenic activity of manure will aid in the development of practices to reduce environmental estrogen accumulation. / Master of Science

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京都大学 / 0048 / 新制・論文博士 / 博士(農学) / 乙第13101号 / 論農博第2847号 / 新制||農||1051(附属図書館) / 学位論文||H29||N5033(農学部図書室) / (主査)教授 廣岡 博之, 教授 松井 徹, 准教授 熊谷 元 / 学位規則第4条第2項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

Submitted by Ubirajara Cruz (ubirajara.cruz@gmail.com) on 2017-05-11T15:29:24Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Marilia_Carvalho.pdf: 854756 bytes, checksum: 8b327668b2ab1f2615a6a8f3f2dd9304 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-05-12T12:17:06Z (GMT) No. of bitstreams: 2 Dissertacao_Marilia_Carvalho.pdf: 854756 bytes, checksum: 8b327668b2ab1f2615a6a8f3f2dd9304 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-05-12T12:17:06Z (GMT). No. of bitstreams: 2 Dissertacao_Marilia_Carvalho.pdf: 854756 bytes, checksum: 8b327668b2ab1f2615a6a8f3f2dd9304 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-05-20 / Sem bolsa / Esta dissertação está apresentada na forma de um artigo científico cujo o objetivo de estudo foi determinar a prevalência e incidência de Leucose Enzoótica Bovina (LEB) e Diarréia Viral Bovina (DVB), avaliando a dinâmica de transmissão de enfermidades virais em um centro de recria de novilhas leiteiras submetidas a Pastoreio Racional Voisin (PRV). Estas doenças virais têm grande impacto econômico na criação de bovinos leiteiros levando a queda na produção leiteira e imunossupressão pré-dispondo as novilhas a outras infecções. Para detecção de anticorpos contra Leucose Enzoótica Bovina foi realizado um teste de Imunodifusão em Agar Gel (IDGA) comercial. Nos animais soropositivos, foi realizado o leucograma pareado para a determinação de linfocitose persistente (LP). Para a determinação da prevalência do vírus da Diarréia Viral Bovina, foi realizado o teste de vírus neutralização(VN) com cepa citopática NADL (BVDV-1). A detecção de possíveis animais persistentemente infectados (PI) foi utilizada a técnica de RT-PCR. O lote experimental era composto por 179 animais de raças leiteiras e cruzas, divididos em cinco lotes por faixa etária e peso, não fixos, submetidos ao manejo pelo método do Pastoreio Racional Voisin (PRV). As taxas de prevalência iniciais para LEB e DVB foram de 3,91% e 51,98% respectivamente. Foram calculadas taxas de incidência para as duas doenças resultando em 21,51% para LEB e 11,77% para DVB. A dinâmica de transmissão da Diarréia viral Bovina foi relativamente, lenta justificada pela ausência de animais PI dentro do lote experimental. Constatou-se que dos 39 animais positivos para LEB ao final do período experimental, dez apresentaram linfocitose persistente (LP), ou seja, uma prevalência de 25,64%, considerada alta, elevando os riscos de transmissão do vírus para outros animais dentro do centro de recria. A intensificação do manejo das novilhas dentro do centro de recria aumenta os riscos de disseminação de destas doenças. Como formas de controle destas enfermidades no rebanho, sugeriu-se a identificação de animais LEB positivos com estratégias de manejo e utensílios separados dos animais negativos; quanto a DVB como não foram encontrados animais persistentemente infectados no rebanho a vacinação das novilhas na idade reprodutiva deve ser preconizada reduzindo o risco da ocorrência de animais PI nas unidades de produção após o retorno dos animais e evitando problemas reprodutivos. / This work is presented in the form of a scientific paper whose objective of the study was to determine the prevalence and incidence of Bovine Leukosis (LEB) and bovine viral diarrhea (DVB), evaluating the dynamics of transmission of viral diseases in a center for rearing dairy heifers Pastoreio Racional Voisin (PRV). These viral diseases have a significant economic impact on the creation of dairy cattle leading to reduced milk production and pre-arranging the heifers to other infections immunosuppression. For antibodies against Bovine Leukosis was carried out immunodiffusion test Agar Gel (AGID) commercial. In seropositive animals, was held the WBC paired for the determination of persistent lymphocytosis (PL). To determine the prevalence of bovine viral diarrhea virus, we performed the virus neutralization test (VN) with cytopathic strain NADL (BVDV-1). The detection of possible persistently infected (PI) animals was used for RT-PCR. The experimental plot consisted of 179 animals of dairy breeds and crosses, divided into five lots by age and weight, not fixed, subject to management by the method of grazing Rational Voisin (PRV). The initial prevalence rates for LEB and DVB were 3.91% and 51.98% respectively. Incidence rates were calculated for the two conditions resulting in LEB 21.51% and 11.77% for DVB. The dynamic transmission of bovine viral diarrhea was relatively slow explained by the absence of PI animals within the test batchIt was found that out of 39 animals positive for LEB the end of the trial, ten presented persistent lymphocytosis (LP), or a prevalence of 25.64%, which is considered high, increasing the risk of virus transmission to other animals within the center recreates. Intensified management of heifers in the center of growing increases the risk of spread of these diseases As ways to control these diseases in the herd, suggested the identification of positive LEB animals with separate management strategies and tools of negative animals; as the DVB as vaccination of heifers in the reproductive age persistently infected animals in the herd were not found to be recommended, reducing the risk of occurrence of animal IP in production units after the return of animals and avoiding reproductive problems.