Orientador: Gilson Hélio Toniollo / Coorientador: Márcia Rita Fernandes Machado / Banca: Marion Burkhardt de Koivisto / Banca: Benedito Dias de Oliveira Filho / Banca: Lindsay Unno Gimenes / Banca: Tais Harumi de Castro Sasahara / Resumo: O objetivo geral deste estudo foi descrever as respostas inflamatórias de jumentas Pêga ao sêmen fresco (MN), sêmen extendido (SE) e sêmen congelado (SC) de jumento, tendo como objetivo específico correlacionar esses achados com os baixos índices de gestações de jumentas Pêga por inseminação artificial (IA) com SC. Foram avaliadas diariamente 25 jumentas, através de rufiações, palpações transretais e exames ultrassonográficos durante maio de 2012 a março de 2013. Foram observados 234 estros, sendo 33 (14,22%) estros anovulatórios e 11 (4,74%) duplas ovulações; o comprimento do ciclo estral foi de 28,88±0,95 dias, com sinais externos de estro de 4,50±0,35 dias e o tempo de ovulação em relação ao término do estro de 8,0±2,45 horas entre as 25 jumentas. Diante dos resultados obtidos, as jumentas não apresentaram sazonalidade reprodutiva. Selecionaram-se 12 jumentas ao acaso, as quais foram biopsiadas em diestro para avaliação reprodutiva e nos ciclos estrais subsequentes, as mesmas foram submetidas à uma das técnicas: monta natural (MN), IA-SE e IA-SC próximo à ovulação, realizando-se biópsia endometrial 10 horas após cada evento. Os fragmentos foram conservados em formalina a 10%, solução de Bouin e solução de glutaraldeído tamponado a 10% para análise em microscopia de luz (ML), microscopia eletrônica de varredura (MEV) e microscopia eletrônica de transmissão (MET). As amostras endometriais das jumentas em diestro apresentaram pregas e sulcos do endométrio, células microvilosas e ciliadas, livre de processos patológicos incompatíveis com a reprodução; em estro apresentaram epitélio colunar simples, evidenciando células microvilosas e células ciliadas, com núcleos ovalados, cromatina condensada e, no citoplasma, presença de mitocôndrias, vacúolos e corpúsculos elétron-densos. Formações semelhantes a bolhas estavam presentes nas superfícies das células microvilosas. As glândulas ... / Abstract: The general objective of this study was to describe the inflammatory responses in Pêga Jennies from fresh (NM), extended (ES) and frozen (CS) semen from Jackass. The specific objective was to correlate these responses with low pregnancy rates in Jennies after artificial inseminations (AI) with frozen semen. Twenty five Jennies were daily evaluated by teasing, rectal examination and ultrasound from May 2002 to March 2013. Estrous cycles (n=234) were followed, with 33 (14.22%) non- ovulatory estrous and 11 (4.74%) double ovulations. The estrous cycle length was 32.68±0.76 days, with external heat signs of 4.49±0.23 days and time of ovulation in relationship to end of estrous of 5.68±7.18 hours for Jennies during the observed period. The estrous cycle was 28.88±0.95 days; with external heat signs of 4.50±0.35 days and time to ovulation in relation to end of estrous of 8.0±2.45 hours among 25 Jennies. Thus, considering the obtained results there was no reproductive seasonality. Twelve Jennies were casually selected for biopsy during diestrous being reproductively evaluated and subsequently submitted to one of those: natural mount (NM), AI- ES and AI-CS near the time of ovulations; and then submitted to endometrial biopsy 10 hours latter. Each fragment was maintained in formalin 10%, Bouin solution and Buffer glutaraldehyde 10% for Ligh Microscopy (LM), Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). The endometrial samples from Jennies in diestrous presented folds and endometrial grooves, microvillus cells and ciliated cells, free of process that could compromise reproduction. The estrous samples presented simple columnar epithelium, circled nuclei, condensed chromatin and at the cytoplasm, mitochondrial vacuoles and electro- dense corpuscles. Similar structures to blebs were present at the microvilli cell surface. The endometrial glands were completed by secretions that were shown with major evidence ... / Doutor

Orientador: João Carlos Pinheiro Ferreira / Banca: Eunice Oba / Banca: Fabiana Ferreira Souza / Banca: Anthony Cesar Castilho / Banca: Anibal Ballarotti Nascimento / Resumo: O objetivo desse estudo foi descrever as características do corpo lúteo (CL) durante a luteólise parcial, em diferentes dias do ciclo estral, após desafio com sub-dose de cloprostenol sódico. Para tanto foram realizados dois experimentos que foram descritos em dois artigos. No artigo I, 32 vacas foram submetidas à sincronização da ovulação (Ovsynch + P4), sendo dessas, 28 ovularam entre 24 e 32 horas (h) após último GnRH (Dia 0) do protocolo. No D6 as vacas foram divididas aleatoriamente em três tratamentos (momento 0h): Controle (2 mL, salina, i.m.; n=10), 2XPGF (duas doses de 500 μg de cloprostenol sódico com 2 horas de intervalo, i.m.; n=8) ou 1/6PGF (83,3 μg de clorprostenol sódico, i.m.; n=10). Amostras de sangue e volume luteal, foram coletadas antes dos tratamentos e a cada 8 h durante às 48 h de observações. Também foram coletadas duas biopsias de CL nos momentos 24 e 40 h pós-tratamento. Valores com P < 0,05 foram considerados diferentes estatisticamente. Nos animais do tratamento 2XPGF, a diminuição da concentração plasmática de progesterona (P4) foi observada a partir de 16 h permaneceu diminuindo até 48 h pós-tratamento. No tratamento 1/6PGF foi observada a luteólise parcial, caracterizada pelas menores concentrações da P4 no momento 16 h seguido do aumento de nos momentos seguintes. As marcações de COX-2 e PGDH foram intensas nos momentos 24 e 40 h, porem a marcação para StAR foi intensa somente no momento 40 h. Ainda no tratamento 1/6PGF o volume luteal também diminuiu com 24 h e esse momento foi caracterizado pela menor área das células luteais grandes (LLC), comparada com o Controle, também algumas células positivas para a marcação de FAS e FAS-L. Nos momentos seguintes o volume luteal aumentou assim como a área das células tiveram tendência em aumentar. No artigo II, 39 vacas foram divididas em dois grupos e submetidas a sincronização da ovulação (Ovsynch + P4), sendo dessas ... / Abstract: We aimed describe the CL characteristics during partial luteolysis, in different days of estrous cycle, after cloprostenol sodium low-dose challenge. Therefore, two projects were conduct and describe in two articles. In the first article, 32 cows were submitted to ovulation synchronization (Ovsunch +P4), 28 cows ovulated between 24 and 32 hours (h) after last GnRH (Day 0) of ovulation synchronization protocol, and were used in the project. At D6, cows were randomly divided in three treatments (moment 0): Control (2 mL of saline 0.9%; IM, n=10), 2XPGF (two doses of 500 μg of cloprostenol sodium with 2 hours interval, n=8) and 1/6PGF (83.33 μg of cloprostenol sodium, IM, n=10). Blood samples and luteal volume were collect before treatments and every 8 h, during 48 h of observations. We also collect two CL biopsies at 24 and 40 h post-treatments. The data was considered statically different when P < 0.05. Animals from 2XPGF had progesterone concentration (P4) decreased 16 h after treatment and it remain low until 48 h after treatment. In 1/6PGF, we observe a partial luteolysis, with decrease on circulating P4 and luteal volume 16 h, after that, we observe an increase in the next moments. The COX-2 and PGDH staining were more intense at 24 and 40 h, however the staining for StAR was intense only on 40 h moment. Still in 1/6PGF treatment, the luteal volume also decrease after 24 h and this moment was characterized by large luteal cells (LLC) minor area than Control, also some positive cell for FAS and FAS-L staining. In the next moments, luteal volume increase as well as cell area tend increase. In the second article, 39 cows were submitted to ovulation protocol (last GnRH = D0), 34 ovulated between 24 and 32 h after GnRH of protocol. The cows were synchronized in a way that data collect were at the same day. Each estrous cycle day were submitted in the same 3 treatments described in Article I: D7 (n=5/2XPGF, n=5/1/6PGF e n=6/control) and ... / Doutor

Recently, several glycerophosphocholine biomarkers for multiple sclerosis were discovered in serum, plasma, and cerebrospinal fluid; little is known, however, about brain glycerophosphocholine metabolism during multiple sclerosis despite evidence that lysophosphocholines can elicit demyelination experimentally. Using a lipidomics approach, glycerophosphocholine metabolites in the hippocampus and temporal-parietal-entorhinal cortex of female C57BL/6J mice subjected to experimental autoimmune encephalomyelitis (a mouse model of multiple sclerosis) were quantified and compared to metabolite levels in healthy mice. To control for potential hormonal regulation, glycerophosphocholine metabolites from these same regions were quantified across the estrous cycle in healthy female N5 C57BL/6J x C3h/HeJ mice. I found that several critical glycerophosphocholine metabolites were significantly decreased over the course of experimental autoimmune encephalomyelitis in both brain regions, although the hippocampus was more affected compared to the temporal-parietal-entorhinal cortex. Similarly, hippocampal glycerophosphocholine metabolism was more responsive to fluctuations in female sex hormones than the cortex. Overall, these results suggest that glycerophosphocholine metabolism differs not only between brain regions, but also between conditions, namely experimental autoimmune encephalomyelitis and the estrous cycle.

At the author’s request, the abstract has been removed due to the confidential nature of the thesis. It will be added once the embargo period has passed.

There is a very few papers to provide an overview of the characteristics of the estrous cycle, the relationship of the estrous cycle to physiological manifestations such as the pH of the vaginal environment, as well as the dynamics of the vaginal microbiota in wild mice. The aim of this thesis is to contribute to the understanding of the dynamic relationship between external influences and the physiology of the female reproductive system, to develop a reliable methodology for measuring the pH of the vaginal microenvironment in mice as well as to quantify the overall abundance of some bacterial taxons by comparing sequencing and qPCR methods. The results suggest that the physical presence of the male in the cage has the most significant effect on the prolongation of the estrus phase, in contrast to non-significant olfactory stimulation of the urine. Fluctuation in the pH of the vaginal environment have also been shown to be cyclic, and the qPCR method shows that the composition of the vaginal microbiota, during the estrus phase, differs significantly from other phases of the estrous cycle, as we confirmed by 16S rRNA sequencing. Thus, these results provide a comprehensive view of the variability of the estrous cycle with an emphasis on the variability of the vaginal microbiota and the change in the...

No description available.

Estrous synchronization (ES) protocols enhances profitability of beef cow-calf operations by improving percentage of cows pregnant early in the breeding season and increasing kg of calf weaned per cow exposed. Many factors, however, influence the fertility of cattle enrolled to ES protocols. The overall goal of this work was to determine the influences of estrus expression and timing of artificial insemination (AI), as well as temperament on fertility of Bos taurus beef females exposed to ES protocols. Multiparous cows (n = 1,676) were enrolled to the 7-d CO-Synch+ Controlled internal drug releasing device (CIDR) protocol and cows expressing estrus were inseminated. Cows not expressing estrus received a gonadotropin-releasing hormone injection and were either inseminated immediately or delayed by 8 hours. Pregnancy rates were greater for cows expressing estrus to the protocol. Delaying AI for 8 hours in cows that failed to express estrus did not improve pregnancy rates. The effects of temperament on fertility of beef heifers enrolled to the 7-d CO-Synch+CIDR protocol were investigated. Heifers (n = 297) had temperament assessed by chute score and exit velocity, and classified as adequate or excitable. Hair and blood were collected for cortisol evaluation. Pregnancy rates were greater for adequate temperament heifers. Circulating cortisol concentrations were greater for excitable heifers, and overall plasma and hair cortisol concentrations were reduced from the start of the protocol to the end. Heifers with adequate temperament have improved fertility and ES protocols acclimated heifers and reduced plasma and hair cortisol concentrations due to handling. / Master of Science / Infertility is a major issue that affects profitability of beef cow-calf producers. Estrous synchronization (ES) protocols are a tool that allow artificial insemination (AI) without estrus detection and increases the proportion of females pregnant at the start of the breeding season, providing a strategy to improve profitability. However, females enrolled in the ES protocol and that fail to express estrus have reduced pregnancy rates when compared to females expressing estrus. Furthermore, beef females with excitable temperament have reduced fertility when compared to females with adequate temperament. The effects of delaying insemination to 8 hours post injection of gonadotropin-releasing hormone for animals failing to express estrus during ES were determined in beef multiparous cows. Cows expressing estrus had greater pregnancy rates compared to cows not expressing estrus, and delayed insemination did not improve pregnancy rates. To determine the effects of temperament on fertility heifers were enrolled in an ES protocol and had temperament determined as adequate or excitable based on chute score and exit velocity. Hair and blood samples were collected for cortisol evaluation. Excitable heifers had reduced pregnancy rates and greater circulating cortisol concentrations, but hair cortisol did not differ between temperaments. Overall cortisol profiles indicate that heifers became acclimated to handling during ES protocol. Development of ES that maximizes estrus expression prior to AI and selection of cattle with adequated temperament can enhance profitability of cow-calf operations.

The objective was to evaluate the insulin-leptin interactions in grazing mares and their impact on aspects of reproductive function using a series of four studies. Fourteen mares were first examined, 10 grazing and 4 confined to stalls in four 22-h studies in April, August, October (2005) and January (2006). There was a positive relationship between insulin and leptin concentrations (r = 0.50; P &lt; 0.001). The second study used 24 mares maintained at pasture and fed supplements rich in either FF, or SS, or pasture forage. Nonstructural carbohydrate (NSC) content of forage was correlated to insulin concentration (r = 0.55; P &lt; 0.01). Insulin was associated with leptin (r = 0.55; P &lt; 0.001) and progesterone (r = 0.48; P &lt; 0.001). In the third study nine mares adapted to FF, SS or forage only underwent two FSIGT tests; once during the luteal phase and again during the follicular phase of the estrous cycle. Minimal model analysis was used to describe insulin sensitivity (SI), glucose effectiveness (Sg), acute insulin response to glucose (AIRg) and the disposition index (DI). SI was lower (P &lt; 0.001) in the luteal phase (3.1 &plusmn; 0.6) compared to the follicular phase (5.0 &plusmn; 0.6). In the fourth study 15 mares had their follicular fluid collected during the luteal and follicular phase of the estrous cycle. Insulin concentration (ln (x+1)) was 52% higher (P &lt; 0.01) in large (&gt; 25 mm) follicles (1.4 ± 0.1 mIU/L) than either medium (16 to 25 mm) or small (le; 15 mm) follicles (0.9 &plusmn; 0.1; 0.9 &plusmn; 0.1 mIU/L, respectively) irrespective of estrous cycle phase. A correlation was observed between follicular fluid (FFL) leptin and plasma leptin (r = 0.30; P &lt; 0.001). A similar relationship was observed between FFL insulin and plasma insulin (r = 0.25; P &lt; 0.001). Plasma insulin and leptin were positively associated (r = 0.45, P &lt; 0.0001), along with FFL insulin and FFL leptin (r = 0.46, P &lt; 0.0001). Both leptin and insulin were affected by season and this observation contradicts the use of single sample analysis for determining detrimental concentrations of these hormones. / Ph. D.

In CBA/NJ mice, splenic natural killer (NK) cell activity varies with stages of estrous. Susceptibility of ICR mice to intravaginal inoculation of herpes simplex virus type 2 (HSV-2) decreases with age. Susceptibility of female ICR and CBA/NJ mice to HSV-2 inoculated intravaginally and intraperitoneally was examined during the estrous cycle. In cycling ICR mice, greatest susceptibility to intravaginal inoculation was observed during diestrous and the least during metestrous. CBA/NJ mice were most susceptible to intravaginal inoculation of HSV-2 during diestrous. ICR mice were ovariectomized to mimic diestrous and found to be highly susceptible to intravaginal inoculation at all virus doses. No difference in susceptibility among phases of the estrous cycle was seen following intraperitoneal inoculation.

O trabalho teve por objetivo estudar o ciclo ovariano de fêmeas de gato-mourisco (Herpailurus yaguaroundi), mantidas em cativeiro, utilizando técnicas de extração e quantificação de metabólitos fecais de hormônios esteróides sexuais e corticosterona. A mensuração dos metabólitos fecais de estradiol, progesterona e cortisol foi efetuada por meio da técnica de radioimunoensaio (RIE). Foram utilizadas oito exemplares fêmeas, mantidas na Fundação Parque Zoológico de São Paulo, São Paulo. As colheitas foram realizadas cinco vezes por semana durante o período de nove meses. As amostras foram acondicionadas individualmente em sacos plásticos e mantidas em freezer a - 20°C. A mediana da duração do ciclo estral (n=33) de gato-mourisco, definido por dois picos consecutivos de metabólitos fecais de estradiol, encontrada no presente estudo, foi de 24 dias; e a duração média±EPM do estro (n=41) foi de 8,65±0,38 dias. O valor da mediana dos valores basais para os metabólitos de estradiol foi de 4,67ng/g de fezes, e dos valores de pico 87,32ng/g de fezes. Para as progestinas os valores encontrados para mediana foram de 0,75ug/g de fezes e 7,49ug/g de fezes para valores basais e de pico respectivamente. As variações no perfil de progestinas sugeriram não terem ocorrido ovulações espontâneas. Não foi encontrada correlação entre progestinas e corticosterona durante os períodos de gestação, lactação e após a retirada dos filhotes. / The aim of this study was analyze the ovarian cycle of captive jaguarundi (Herpailurus yagouaroundi) trough fecal steroid. Radioimmunoassay (RIA) was used for the measurement of estradiol, progesterone and cortisol metabolites in feces. Fecal samples were collected five days per week from eight females kept at Fundação Parque Zoológico de São Paulo during nine months. These samples were placed into zip-lock bags and stored in freezer at - 20°C. The median of estrous cycle length, defined by two consecutive peaks of fecal estradiol metabolites, found in this study was 24 days (n=33 cycles) and the mean (±SEM) of estrous duration was 8,65±0,38 days (n= 41 cycles). The median concentrations of fecal estradiol metabolites were 4,67ng/g of feces and 87,32ng/g of feces for basal and peak values respectively. Progestins median found was 0,75ug/g of feces for basal and 7,49ug/g of feces for peak values. The fluctuations in progestins profiles suggested that spontaneous ovulation did not occur. No correlations were found between progestins and fecal corticosterone during pregnancy, lactation and kitten removal.