Dissertations / Theses on the topic 'Semen parameter'

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2018-04-18T16:43:21Z No. of bitstreams: 1 texto completo.pdf: 1119071 bytes, checksum: 3dad9c3c8b4da25dfaf3676d6dcc7e69 (MD5)<br>Made available in DSpace on 2018-04-18T16:43:21Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1119071 bytes, checksum: 3dad9c3c8b4da25dfaf3676d6dcc7e69 (MD5) Previous issue date: 2017-09-04<br>Coordenação de Aperfeiçoamento de Pessoal de Nível Superior<br>O uso generalizado da inseminação artificial (IA) contribuiu grandemente para o sucesso da indústria de suínos, por meio do auxílio e disseminação do progresso genético. Atualmente, reprodutores suínos jovens são selecionados para IA com base em seus valores genéticos para características de produção e, a seleção de reprodutores para características de sêmen, como volume, concentração, motilidade e morfologia, bem como para menor variação intra- reprodutor na sua produção e qualidade, ainda não é uma prática comum. Esta seleção é importante para melhorar o desempenho dos reprodutores nas estações de IA, cujo objetivo é maximizar o número de doses inseminantes produzidas por cada ejaculado. A estimação de parâmetros genéticos e quantificação da variação genética para características de sêmen e para variação intra-reprodutor permitem analisar se essas características devem ser incluídas nos objetivos do melhoramento. Além da estimação de parâmetros genéticos para fins de seleção, o interesse em estudar os processos moleculares e os mecanismos genéticos que afetam as características de sêmen está aumentando nos últimos anos. Os estudos de associação genômica ampla (GWAS) são comumente usados para identificar polimorfismos de base única (SNPs) associados a loci de características quantitativas (QTL) com maiores efeitos. O GWAS em passo único ponderado (WssGWAS) é um método que permite a estimação de efeitos de SNP utilizando informações de todos os animais genotipados, fenotipados e com pedigree na população. Expandindo as fronteiras dos estudos de reprodução em suínos, outro campo importante a ser explorado em programas de melhoramento é a fertilidade dos reprodutores. As características reprodutivas, como a duração da gestação (GL), o número total de leitões nascidos (TNB) e nascidos mortos (SB) são algumas características-chave para a produção eficiente de suínos. Devido às baixas ou moderadas herdabilidades para essas características, é importante identificar todos os fatores que as influenciam e incluir esses fatores nos modelos de avaliação genética. Os efeitos do reprodutor cujo ejaculado foi utilizado para inseminar a matriz e do ejaculado são dois desses fatores importantes que têm o potencial de melhorar os modelos tradicionais utilizados nas avaliações genéticas das características reprodutivas. Dentre os elementos que controlam o tamanho da leitegada, as taxas de fertilização e de sobrevivência pré-natal podem ser influenciadas pelo reprodutor, devido às diferenças genéticas na capacidade de fertilização relacionadas à qualidade do sêmen e/ou à contribuição genética do reprodutor para a viabilidade do embrião. Nesse contexto, os objetivos gerais com este estudo foram 1) estimar os parâmetros genéticos para qualidade e quantidade de sêmen, bem como para a variação intra-reprodutor para essas características; 2) identificar regiões de QTL e genes candidatos associados a características de sêmen por meio do WssGWAS e, subsequentemente, realizar análises de redes gênicas para investigar os processos biológicos compartilhados por genes identificados em diferentes linhas de suínos e 3) estimar parâmetros genéticos para o efeito do reprodutor na GL, TNB e SB e avaliar a inclusão dos efeitos do reprodutor e do ejaculado nos modelos de avaliação genética dessas características. Os resultados desta tese mostraram estimativas moderadas de herdabilidade e correlações genéticas favoráveis entre características de sêmen, indicando que a seleção de reprodutores para essas características pode resultar em razoável progresso genético. Além disso, variação genética relevante foi encontrada para a variabilidade intra-reprodutor para essas características, revelando a possibilidade de seleção de reprodutores para uma menor variação na qualidade e produção de sêmen. Os resultados do WssGWAS apontaram regiões relevantes de QTL que explicaram grandes proporções da variância genética (até 10,8%) para as características de sêmen em vários cromossomos suínos, confirmando a suposição de complexidade genética dessas características. Esta identificação foi possível com o baixo número de animais com fenótipos e genótipos, devido à escolha apropriada do método. Os genes candidatos SCN8A, PTGS2, PLA2G4A, DNAI2, IQCG, LOC102167830, NME5, AZIN2, SPATA7, METTL3 e HPGDS foram identificados associados às características de sêmen nas regiões de QTL identificadas para as linhas de suínos avaliadas. A análise de redes gênicas mostrou genes candidatos encontrados para diferentes linhas de suínos compartilhando caminhos biológicos que ocorrem nos testículos de mamíferos. No que diz respeito à fertilidade do reprodutor, os resultados mostraram que há variação genética devido ao efeito do reprodutor em GL, TNB e SB; e o modelo com inclusão de efeitos de ambiente permanente e genéticos do reprodutor, além do efeito do ejaculado, mostrou o melhor ajuste para os dados. Esta tese resultou em informações científicas importantes e inovadoras na área de reprodução em machos, o que contribuirá para aumentar o conhecimento ainda escasso sobre a seleção genética e a arquitetura genômica de características de qualidade de sêmen e de fertilidade em reprodutores suínos.<br>The widespread use of artificial insemination (AI) has greatly contributed to the success of the pig industry by assisting and disseminating the genetic progress. Currently, young boars are selected for AI based on their breeding values for production traits and selecting boars for semen traits, such as volume, concentration, motility and morphology, and for low variation in semen quality and production is still not a common practice. This selection is important for better performance of boars at AI stations, whose objective is to maximize the number of insemination doses produced by each ejaculate. The estimation of genetic parameters and the quantification of genetic variation for semen traits and within-boar variation allow an analysis of whether these traits should be included in the breeding goal. Besides the estimation of genetic parameters for selection purposes, the interest in studying the molecular processes and genetic mechanisms affecting semen traits is increasing in recent years. Genome-wide association studies (GWAS) are commonly used to identify single nucleotide polymorphisms (SNPs) associated with quantitative trait loci (QTL) with major effect. The weighted single-step GWAS (WssGWAS) is a method that allows estimation of SNP effects using information from all genotyped, phenotyped and pedigree animals. Expanding the frontiers of reproduction studies in pigs, another important field to be explored in breeding programs is the boar fertility. Reproductive traits, such as gestation length (GL), total number of piglets born (TNB) and stillborn (SB) are some of the bottleneck traits for efficient pig production. Because of the low to moderate heritabilities for these traits, it is important to identify all factors influencing them and to include these factors in the genetic evaluation models. The service sire (boar which ejaculate dose was used to inseminate the sow) and ejaculate effects are two of those important factors that have the potential to improve the traditional models used in the genetic evaluations of reproductive traits. Among the elements controlling the litter size, the fertilization rate and prenatal survival rate might be influenced by the service sire due to genetic differences in the capacity of fertilization, which is related to sperm quality and/or the boar genetic contribution to viability of the embryo. In this context, my overall aims were 1) to estimate genetic parameters for semen quality and quantity traits, as well as for within-boar variation of these traits; 2) to identify QTL regions and candidate genes associated with semen traits through a WssGWAS and, subsequently, to perform gene network analyses to investigate the biological processes shared by genes identified in different pig lines and 3) to estimate genetic parameters for service sire on reproductive traits GL, TNB and SB and to evaluate the inclusion of service sire and ejaculate effects in the genetic evaluation models of these traits. The results of this thesis showed moderate estimates of heritability and favorable genetic correlations between semen traits, indicating that boar selection for these traits could make reasonable genetic progress. In addition, relevant genetic variation was found for within-boar variability of these traits, revealing the possibility of selection of boars for reduced variation in semen quality and production. Results from WssGWAS pinpointed relevant QTL regions explaining high proportions of genetic variance (up to 10.8%) for semen traits in several pig chromosomes, confirming the assumption of genetic complexity of these traits. This identification was possible with low number of animals having both phenotypes and genotypes due to the appropriate choice of the method. Candidate genes SCN8A, PTGS2, PLA2G4A, DNAI2, IQCG, LOC102167830, NME5, AZIN2, SPATA7, METTL3 and HPGDS were identified associated with semen traits in the QTL regions identified for the pig lines evaluated. The gene network analysis showed candidate genes found for different pig lines sharing biological pathways that occur in mammalian testes. Regarding boar fertility, the results showed that there is genetic variation due to service sire effect on GL, TNB and SB; and the model with inclusion of permanent environmental and genetic effects due to service sire, in addition to ejaculate effect, showed the best fit to the data. This thesis resulted in important and innovative scientific information on male reproduction field in pigs, which will contribute to increase the still scarce knowledge about genetic selection and genomic architecture of boar semen quality and fertility traits.

This dissertation focuses on the effect of breeding on the myocardium of stallions using cardiac troponin I (cTnI) as the preferred blood biomarker for detection of myocardial cell injury. Haematological, echocardiographic and heart rate (HR) parameters are also reported. Fourteen clinically healthy phantom-trained stallions were assessed. Cardiac troponin I concentration was determined pre-semen collection and at 4, 6, 12 and 24 hours following semen collection. Predictors that were measured included mean HR during each stage of semen collection, maximum HR, area under the curve for HR and fractional shortening (FS). Pre-semen collection cTnI concentrations were within reported reference ranges for all stallions. Following semen collection, cTnI concentration was significantly increased at 4 and 6 hours post-semen collection. Results also suggest that the HR of stallions peak during mounting of the phantom. Five stallions failed to show an expected increase in FS immediately post-semen collection. No association was identified between the measured predictors and increased cTnI concentration. This study concluded that cTnI concentration may increase in stallions following semen collection although the clinical significance of this observed increase in healthy stallions is unclear without histopathological evaluation of the myocardium. No conclusion regarding FS immediately post-semen collection could be drawn from results in this study. The research reported in this dissertation can serve as a reference for future studies investigating cTnI concentrations in stallions.<br>Dissertation (MSc)--University of Pretoria, 2010.<br>Companion Animal Clinical Studies<br>unrestricted

The rationale of this thesis was to examine and extend knowledge of boar semen quality, its evaluation and factors which influence quality; with the objective of improving the effectiveness of pig AI. The literature review led to the conclusion that the development of a semen evaluation technique which describes the overall quality of the sperm, correlates well with most semen quality parameters, and most important, applicable at farm level would be a valuable advance. In addition knowledge is needed of the factors affecting semen quality and in particular better semen dilution techniques. Studies on the effect of various factors on semen quality showed: i. With two boar lines (purebred, PB) and (crossbred, CB), CB gave higher total sperm counts (P<0.05), but PB gave better farrowing rates (87% V 78%, P<0.05), ii. Semen collected in Autumn/Winter had higher total sperm counts than semen collected in Spring/Summer (66.4 V 47.8 x 10⁹, P<0.001), iii. General semen quality did not change significantly over 48 hours in the study, iv. Pooling of semen had no effect on semen quality. Semen dilution by a controlled 'Anti-shock' procedure gave a higher proportion of litters with 11 or more piglets born than semen diluted by the standard procedure (78% vs 62%, P<0.05). Sperm motility was affected when, in Kiev diluent, glucose (G) was replaced by fructose (F). Motility was F vs GF vs G: 84.8%; 84.1; 81.0, but by day-3 was 27.7, 60.2 and 62.0 (P<0.01). No significant differences were found between diluents in AI sow fecundity, probably due to the limited numbers of sows. It is concluded that although farrowing rate and litter size are the ultimate indices of semen quality, good correlations with <I>in vitro</I> techniques have yet to be established. The 'swim-up' test is suggested as the most suitable on farm <I>in vitro</I> semen assessment, and that the 'Anti-shock' dilution technique has commercial promise. Fructose should be the usual sugar used in the diluent.

Objectives of this research were to estimate heritabilities of scrotal circumference and semen traits, and genetic correlations among these traits and birth weight. Line 1 Hereford bulls (n = 841), born in 1963 or from 1967 to 2000, were selected for use by USDA-ARS at Miles City, Montana or for sale. Semen was collected by electro-ejaculation when the bulls were approximately one year of age (mean = 446d) and all samples were evaluated by one person. Traits analyzed were scrotal circumference, color, volume, concentration, swirl, motility, and percents normal, live, abnormal heads, abnormal mid-pieces, proximal distal droplets, bent tails, coiled tails, distal proximal droplets, and primary and secondary abnormalities. Data were analyzed using MTDF-REML. Models included fixed effects for contemporary group, age of dam, age of bull at evaluation, inbreeding of the bull and his dam, and random animal, maternal, permanent maternal environmental, and residual effects. Heritability estimates for scrotal circumference, color, volume, concentration, swirl, motility, and percents normal, live, abnormal mid-pieces, proximal distal droplets, coiled tails, and primary and secondary abnormalities were 0.57, 0.15, 0.09, 0.16, 0.21, 0.22, 0.23, 0.34, 0.17, 0.34, 0.30 0.34, and 0.29, respectively. Estimates of genetic correlations between birth weight and scrotal circumference, color, volume, concentration, swirl, motility, and percents normal, live, abnormal mid-pieces, proximal distal droplets, coiled tails, and primary and secondary abnormalities were 0.36, 0.60, 0.07, 0.58, 0.44, 0.21, 0.20, 0.34, -0.03, -0.52, -0.20, -0.25, and 0.05, respectively. The moderate estimates of heritability for many of the traits indicate potential for favorable selection response. Positive genetic correlations between birth weight and majority of the traits suggest selection to reduce birth weight may compromise semen traits. However, for most traits the expected correlated responses are small. Desirable genetic correlations among scrotal circumference and semen traits suggest selection for one trait would not compromise the other semen traits. Expected correlated responses in semen traits to selection for increased scrotal circumference appear favorable.

Since the main ambition of the European community is to create a Single Market among the European countries, a unified set of structural design codes recently became mandatory to use. These standards, the Eurocodes, contain Nationally Determined Parameters (NDPs) that have to be fixed by the National Standard Bodies in the various countries. The investigation of the adopted values for Eurocode 2 in seven European States has been carried out i  this thesis for quantifying the differences that arise from the selected values of these parameters. The aim of this investigation was to find out which countries are advantaged or disadvantaged by the choice of the value of these parameters. The analysis has been limited to the Part 1-1 of Eurocode 2 that concerns “General Rules for Buildings” and the most important parts of Eurocode 0 and 1. The investigated countries were Denmark, Finland, France, Germany, Italy, Sweden and the United Kingdom. Even if these countries represent only seven out of 27 EU States, this comparison gives a good view of the European situations since they cover nearly half of the continental concrete consumption. The analysis includes a theoretical comparison of the national choices of all the 170 national parameters as well as their influence on the more important formulas contained in the Code. Five practical case studies concerning the design and the verification of some structural elements have been carried out in order to study the differences in real and common design situations. Through all the work, the different choices have been compared to the recommended values proposed by the Eurocode that have been used as a mean of comparison. Due to the large number of parameters and the complexity of the problem it has not been possible to estimate and quantify general trends of differences for the countries although Finland, France, Italy and the United Kingdom have been found to have, in general, more disadvantageous choices than the other States. Moreover, Denmark and Germany show variable trends (both above and below the recommended values) while Sweden is always close to the recommended values. The results that have been found show that still a lot of work and research has to be done in order to achieve an even set of structural standards for the design of concrete structures. A list of more important national parameters has been presented as well as a proposal to convert some national parameters to fixed ones. The analysis and the results that have been obtained give details and indications about the future works that need to be done for decreasing the differences among the European countries.

Czech Republic is one of the world's leading producers of the poppy seeds. Concerning the growing field area of poppy, the Czech Republic is lead producer of the poppy seeds at the international level (Vašák, 2010; Cihlář, 2012). Poppy seed is very sensitive on many internal and external factors during cultivation, especially in the beginning of germination and emergence. At present, there are not registered preparations available for seed dressing for poppy seed in the Czech Republic (Prokinová, 2006). However, there has been done many experiments based on testing of natural substances (eg. humates and plant extracts), microorganisms and especially the application of soil fungi in seed treatment recently The aim of this study is to evaluate the effect of selected regional varieties of poppy cultivation and seed treatment to yield parameters of poppy seed both individually and in combination,. For the thesis was essential to describe the suitability of varieties for growing in organic and integrated agriculture in relation to the efficiency of production. Evaluation of the seed quality of the varieties was carried out in the field as well as under laboratory conditions. Before sowing, seeds were treated by preparations Gliorex and TS Osivo, untreated variant has been used as a control. During plant cultivation were monitored shape of the plants, the degree of infestation, weed control and other treatments based on natural substances. Neem-Azal was used as the pest regulator. The second variant was treated by foliar application of TS Květa. Seeds harvested from variants of our trial were tested under laboratory conditions for comparison of differences between varieties. The tests also revealed the seed infestation and what percentage of the seed is viable. The last evaluation was carried out on the basis of the weight of thousands of seeds (HTS), which reflects the yield determined by the selected seed treatment.

U okviru ove doktorske disertacije ispitani su i utvrđeni optimalni uslovi za proces lju&scaron;tenja semena suncokreta visokooleinskog tipa, kao sirovine za proizvodnju ulja. Sprovedena istraživanja su dala podatke o optimalnim vrednostima sadržaja vlage u semenu i pritiska vazduha u lju&scaron;tilici na osnovu kojih je utvrđen model za lju&scaron;tenje semena. Sprovedenim istraživanjima obuhvaćeno je i ispitivanje uticaja promenljivih tehnolo&scaron;kih parametara, sadržaja ljuske u opsegu od 0 do 20% i sadržaja nečistoća u opsegu od 0 do 10% na sastav i kvalitet hladno presovanih ulja i pogače, prinos ulja, oksidativnu stabilnost, senzorski kvalitet i boju hladno presovanih ulja semena suncokreta visokooleinskog tipa.Dobijeni rezultati su obrađeni primenom savremenih klasifikacionih (klaster analiza i analiza glavnih kompomenata) i regresionih hemometrijskih metoda i postavljeni hemometrijski modeli čija bi primena doprinela proizvodnji hladno presovanog ulja vrhunskog kvaliteta i visoke oksidativne stabilnosti. Utvrđeno je da su definisani modeli statistički validni i da se mogu primeniti za optimizaciju tehnolo&scaron;kih parametara u proizvodnji hladno presovanog ulja semena suncokreta visokooleinskog tipa. Validnost postavljenih modela, odnosno kvalitet dobijenih korelacija su procenjeni na osnovu parametara unakrsne validacije.Maksimalna efektivnost lju&scaron;tenja semena suncokreta visokooleinskog tipa može se postići pri optimalnim uslovima tj. sadržaju vlage u semenu 6,0% i pritisku vazduha u lju&scaron;tilici 800 kPa. Hladno presovano ulje semena suncokreta visokooleinskog tipa se, u pogledu sastava i kvaliteta ulja i pogače, prinosa ulja, oksidativne stabilnosti, senzorskog kvaliteta i boje ulja, može proizvoditi od semena koje sadrži 10-20% ljuske i 0-5% nečistoća.<br>In this dissertation the optimal conditions for the dehulling process of high-oleic sunflower seeds, as raw material for oil production, were tested and determined. The conducted studies provided data for the optimal values of moisture content in the seed and the air pressure in the seed dehulling machine, what presented the base for determination of the dehulling model. Also, conducted studies examine the effect of variable technological parameters, such as content of hull (ranging from 0 to 20%) and the impurities content (ranging from 0 to 10%) on the composition and quality of oil and cake, oil yield, oxidative stability, sensory quality and colour of cold pressed high-oleic sunflower oil.The obtained results were analyzed by modern classification (cluster analysis and principal component analysis) and regression chemometrics methods, and chemometric models whose implementation could contribute to the production of cold pressed oil of high quality and high oxidative stability were set. It was found that the defined mathematical models are statistically valid and that they could be used to optimize the technological parameters of production of cold pressed high-oleic sunflower oil. The validity of the set models, that is the quality of obtained correlations, were estimated based on the parameters of cross-validation.The maximum effectiveness of high-oleic sunflower seeds dehulling can be achieved under optimal conditions, i.e. when the moisture content in seed is 6,0% and when the air pressure in seed dehulling machine is 800 kPa. Regarding composition and quality of the oil and cake, oil yield, oxidative stability, sensory quality and color, cold pressed high-oleic sunflower oil could be produced from seed containing 10-20% hull and 0-5% impurities.

Orientador: Anaglória Pontes<br>Banca: Antonio Alberto Nogueira<br>Banca: Edilberto de Araújo Filho<br>Resumo: A infertilidade é definida pela inabilidade de engravidar após 12 meses ou mais de coito regular não protegido. O uso de polivitamínico e polimineral parece influenciar na qualidade seminal. Dados da literatura sobre o uso oral isolado ou combinado desses micronutrientes na melhoria dos parâmetros seminais e eventual fertilidade são controversos e escassos. Analisar os parâmetros seminais de indivíduos inférteis em uso de polivitamínico e polimineral e compará-los com indivíduos normais, comprovadamente férteis sem uso destas substâncias. Foram analisados os parâmetros Seminais de 57 casais inférteis acompanhados no ambulatório de esterilidade do Hospital das Clínicas da Faculdade de Medicina de Botucatu, no período de 2003 a 2007. Nos indivíduos inférteis a análise seminal foi realizada antes e com 90 dias de micronutrientes por via oral os quais foram comparados com 50 indivíduos saudáveis comprovadamente férteis sem uso destas substâncias. A avaliação do sêmen foi feita de acordo com os critérios da Organização Mundial de Saúde - OMS (1999) e morfologia de Kruger et al. (1986). A análise estatística foi feita utilizando os testes t de Student, Mann-Whitney e Wilcoxon, considerando um nível de significância de 5%. Os indivíduos inférteis e os comprovadamente férteis apresentaram similaridade quanto a idade (31,0±5,6 versus 30,3±6,5) (p=0,55) e ao tabagismo (29,8% versus 22,0) (p=0,36). Nos indivíduos inférteis, o uso desses micronutrientes aumentou significativamente a morfologia tanto pelos critérios estabelecidos pela OMS (18,3±9,6 para 22,6±11,8) (p=0,006) e por Kruger (6,9±4,1 para 9,1±5,2) (p=0,002). Verificou-se que os homens inférteis antes do uso de micronutrientes quando comparados aos férteis apresentavam significantemente uma menor concentração de espermatozóides/ml (68,0[37,8;101,2]... (Resumo completo, clicar acesso eletrônico abaixo)<br>Abstract: Infertility is defined as the inability to become pregnant after 12 months or more of regular unprotected intercourse. The use of multivitamin/multimineral supplements seems to influence semen quality. Data on the isolated or combined use of these micronutrients to improve semen parameters and eventual fertility are controversial and scarce. To assess semen parameters in infertile individuals using multivitamin and multimineral supplements in comparison with healthy proven fertile individuals not using these substances. Semen parameters were evaluated in 57 infertile couples followed up in the Sterility Outpatient Clinic of Botucatu Medical School between 2003 and 2007. Semen analysis was performed before and after 90 days of oral micronutrient use in infertile individuals that were compared with 50 healthy proven fertile individuals not using these substances. Semen was evaluated according to the recommendations of the World Health Organization- WHO (1999) and the criteria described by Kruger et al. (1986). Statistical analysis was carried out using Student's t test and the tests of Mann-Whitney and Wilcoxon with significance set at 5%. Infertile and proven fertile individuals showed similar age (31.0±5.6 versus 30.3±6.5) (p=0.55) and smoking status (29.8% versus 22.0) (p=0.36). In infertile individuals, the use of micronutrients significantly improved morphology according to the criteria of WHO (18.3±9.6 to 22.6±11.8) (p=0.006) and Kruger (6.9±4.1 to 9.1±5.2) (p=0.002). Before micronutrient use, infertile individuals compared with fertile males showed lower spermatozoa/ml concentration (68.0[37.8;101.2] versus 96.5[49.0;144.2]) and vitality (85,4±9,2 versus 89.6±6.9) and higher leukocyte count (600.0 [300.0;121.5] versus 350.0[100.0;675.0]). In infertile individuals using multivitamin and multimineral supplements, semen parameters... (Complete abstract click electronic access below)<br>Mestre

CoordenaÃÃo de AperfeÃoamento de Pessoal de NÃvel Superior<br>O presente estudo foi realizado com o objetivo de identificar o perfil das principais proteÃnas do plasma seminal suÃno e suas associaÃÃes com parÃmetros seminais. Amostras de sÃmen foram coletadas de 12 cachaÃos adultos e submetidas à avaliaÃÃo dos parÃmetros seminais (motilidade total, morfologia, vitalidade e porcentagem de cÃlulas com acrossoma intacto). O plasma seminal foi obtido por centrifugaÃÃo e as proteÃnas seminais foram analisadas por eletroforese 2-D e espectrometria de massa. Foram testados modelos de regressÃo usando a soma das intensidades dos spots referentes Ãs mesmas proteÃnas como variÃveis independentes e parÃmetros seminais como variÃveis dependentes (p < 0,05). Cento e doze spots foram identificados nos gÃis do plasma seminal, equivalentes a 39 proteÃnas diferentes. As espermadesinas PSP-I, PSP-II, AQN1, AQN3 e AWN1 representaram 45,2  8 % do total da intensidade de todos os spots detectados nos geis. Outras proteÃnas expressas no plasma seminal suÃno incluem a albumina, proteÃnas do complemento (complement factor H precursor, complement C3 precursor e adipsin/complement factor D), imunoglobulinas (IgG heavy chain precursor, imunoglobulina delta heavy chain membrane bound form, imunoglobulina gamma-chain, Ig lambda chain V-C region PLC3 e CH4 and secreted domain of swine IgM), IgG-binding proteins, epididymal specific lipocalin-5, epididymal secretory protein E1 precursor, epididymal glutathione peroxidase precursor, transferrin, lactotransferrin e fibronectin tipo 1 (FN1). Em funÃÃo das anÃlises de regressÃo, o percentual de espermatozoides com defeito na peÃa intermediÃria foi relacionado com a quantidade de âCH4 and secreted domain of swine IgMâ e FN1 (R2 = 0,58), a proteÃna IgG-binding (R2 = 0,61), o fator H do complemento (R2 = 0,61) e lactaderina (R2 = 0,45). Porcentagem percentual de defeitos de cauda tambÃm foi relacionado com âCH4 and secreted domain of swine IgMâ e FN1 (R2 =0,40), a proteÃna IgG-binding (R2 = 0,34), e lactaderina (R2 = 0,74). A motilidade espermÃtica, por sua vez, teve associaÃÃo com a intensidade dos spots identificados como lactaderina (R2 = 0,48). Em conclusÃo, descreve-se, neste trabalho, o proteoma do plasma seminal suÃno e associaÃÃes significativas entre proteÃnas especÃficas do plasma seminal e parÃmetros seminais. Tais relaÃÃes servirÃo como base para a determinaÃÃo de marcadores moleculares da funÃÃo espermÃtica na espÃcie suÃna.<br>The present study was conducted to identify the major seminal plasma protein profile of boars and its associations with semen criteria. Semen samples were collected from 12 adult boars and subjected to evaluation of sperm parameters (motility, morphology, vitality and percent of cells with intact acrosome). Seminal plasma was obtained by centrifugation, analyzed by 2-D SDS-PAGE and proteins identified by mass spectrometry (electrospray ionization quadrupole-time-of-flight). We tested regression models using spot intensities related to the same proteins as independent variables and semen parameters as dependent variables (p < 0,05). One hundred and twelve spots were indentified in the boar seminal plasma gels, equivalent to 39 different proteins. Spermadhesins PSP-I, PSP-II, AQN1, AQN3 and AWN1 represented 45,2  8 % of the total intensity of all spots. Other proteins expressed in the boar seminal plasma include albumin, complement proteins (complement factor H precursor, complement C3 precursor and adipsin/complement factor D), immunoglobulins (IgG heavy chain precursor, immunoglobulin delta heavy chain membrane bound form, immunoglobulin gamma-chain, Ig lambda chain V-C region PLC3 and CH4 and secreted domain of swine IgM), IgG-binding proteins, epididymal specific lipocalin-5, epididymal secretory protein E1 precursor, epididymal secretory glutathione peroxidase precursor, transferrin, lactotransferrin and fibronectin type 1 (FN1). Based on regression analysis, the percentage of sperm with midpiece defects was related to the amount of CH4 and secreted domains of swine IgM and FN1 (R = 0,58), IgG-binding protein (R = 0,61), complement factor H precursor (R = 0,61) and lactadherin (R = 0.45). The percentage of sperm with tail defects was also related to CH4 and secreted domains of swine IgM and FN1 (R = 0,40), IgG-binding protein (R = 0,34) and lactadherin (R = 0,74). Sperm motility, in turn, had association with the intensities of spots identified as lactadherin (R = 0,48). In conclusion, we presently describe the major proteome of boar seminal plasma and significant associations between specific seminal plasma proteins and semen parameters. Such relationships will serve as the basis for determination of molecular markers of sperm function in the swine species.

Introdução: Em torno de 15% dos casais apresentam o diagnóstico de infertilidade, sendo que 50% se devem a fatores masculinos. Diversos testes de função espermática são propostos para a avaliação da fertilidade, mas o espermograma é o principal exame para o diagnóstico de infertilidade masculina. O valor prognóstico das características seminais como concentração, morfologia e motilidade como marcadores de infertilidade masculina é confundido. A avaliação desses e a classificação quanto à normalidade dos mesmos permanece sendo tema de discussão frequente. Objetivos: Comparar as diferentes classificações de parâmetros seminais (OMS 1999, OMS 2010 e TMSC) e avaliar as características de pacientes em terapia de reprodução humana assistida que se relacionam com esses parâmetros. Métodos: Foi realizado estudo longitudinal, retrospectivo, baseado em revisão de banco de dados para avaliar as características e parâmetros seminais e comparar as mesmas conforme as classificações da OMS de 1999, OMS de 2010 e TMSC de 477 homens submetidos a investigação de infertilidade ou tratamentos de reprodução assistida na clínica Generar – Reprodução Humana, entre 2011 e 2015. Resultados: 401 pacientes tiveram alguma alteração pela OMS 1999, 223 pela OMS 2010 e 200 pela TMSC. O critério que mais alterou a condição de uma amostra seminal de anormal em 1999 para normal em 2010 foi a morfologia. Conclusão: Os parâmetros se tornaram menos rígidos de 1999 para 2010 alterando significativamente a proporção de indivíduos que deixaram de ser classificados como inférteis. A classificação baseada no TMSC pode ser útil na indicação de alguns tratamentos, mas não pode definir um indivíduo como fértil ou infértil em função de não levar em consideração a morfologia espermática.<br>Background: Approximately 15% of couples presenting the diagnosis of infertility, and 50% of cases are due to male factors. Several sperm function testing is proposed for the evaluation of male fertility, but sperm is the first test for the diagnosis of male infertility causes. The prognostic value of the seminal characteristics as concentration, morphology and motility as male infertility markers is often confused. Evaluation of semen parameters and classification for normality remains frequent topic of discussion. Objectives: Compare the different classifications of seminal parameters (OMS 1999, WHO 2010 and TMSC) and evaluate the characteristics of patients treated in assisted reproduction therapy relating to these parameters. Methods: Retrospective study based on chart review evaluated 477 semen samples from men undergoing investigation or infertility treatments in assisted reproduction between 2011 and 2015. Results: 401 patients were considered abnormal by the WHO 1999, 223 by WHO 2010 and 200 for TMSC. The criteria that most changed the classification was sperm morphology. Conclusion: The parameters have become less rigid 1999 to 2010 significantly changing the proportion of individuals who are no longer classified as infertile. The classification based on TMSC can’t define an individual as fertile or infertile regardless due to not take into account the sperm morphology, but may be helpful when it comes to the indication of the intrauterine insemination.

Introduction International guidelines endorse the screening of patients for human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV) and Chlamydia trachomatis before assisted reproductive techniques (ART). At present no such guidelines exists in South Africa. At the Reproductive and Endocrine Unit (referred to as “the Unit”) of Steve Biko Academic Hospital, all patients with unknown HIV status are counselled and a blood sample is collected during the initial visit for automated laboratory based HIV screening. These HIV results are not available before semen samples are processed. Furthermore, patients are not screened for HBV, HCV and Chlamydia trachomatis. Couples attending the Unit are of a low to middle socio-economic status and experience financial constraints. Moreover, automated laboratory based assays are expensive to perform. Rapid testing is a cost effective and practical method from screening patients, with a 20–30 minute result turnover time. Until screening at the Unit is improved, the possible identification of semen characteristics that could indicate HIV infection would be a useful tool. Materials and Methods The following rapid point-of-care assays were evaluated: Determine® HIV-1/2 combo test (n=100), Determine® HBsAg test (n=100), DIAQUICK HCV kit (n=74), and the DIAQUICK Chlamydia trachomatis kit (n=30). For profiling, parameters from a basic semen analysis of HIV-positive males (n=60) were compared with HIV-negative males (n=60). Information pertaining to CD4 count, antiretroviral treatment and plasma viral load of HIV-positive males were analysed. Results From all patients included in the study, 8% tested positive for HIV. The risk of a female being HIV-positive was 3.73 times higher than for males. In the pilot study to explore rapid testing for HBV and HCV, 1% and 1.4% of patients tested positive respectively. When testing for Chlamydia trachomatis 31.3% of females, but no males tested positive. Comparing semen profiles, no significant differences were found between samples from HIV positive and negative males or between HIV positive males categorised by CD4 cell count (p>0.05). For the HIV-positive group with a detectable plasma HIV viral load (>40 copies/ml), a significant difference was observed in the semen viscosity (p=0.0460). Significant differences were noted in the sperm motility (immotile sperm p=0.0456, progressive sperm p=0.0192) of patients receiving antiretroviral (ARV) therapy. Discussion and Conclusion The use of rapid testing is an acceptable and feasible option for improving current screening protocols at the Unit. The absence of definite alterations in the semen characteristics of HIV-positive men further motivates the need for a simpler, point-of-care screening protocol. The prevalence of HBV was lower than that reported in the general population of South Africa and further investigation is needed. Although the sample size was small, HCV prevalence was similar to that of the general population. One third of females tested positive for Chlamydia trachomatis. The methodology used was possibly not appropriate for males. This study highlighted the need for guidelines that address the specialised needs of ART clinics in resource-limited and developing countries with a high HIV prevalence.<br>Dissertation (MSc)--University of Pretoria, 2013.<br>gm2014<br>Obstetrics and Gynaecology<br>unrestricted

Thesis (MScMedSc)--Stellenbosch University, 2013.<br>ENGLISH ABSTRACT: CHAPTER 1 A review of the application of traditional semen parameters for the investigation and diagnosis of male infertility and the role of predictive values in assisted reproductive techniques (ART) is presented. The importance of sperm morphology, with special emphasis on sperm morphology evaluation, is discussed. Also presented is an overview of the physiology of sperm DNA, the process of spermatogenesis, as well as the contribution of the spermatozoon to the embryo. The different causes of sperm DNA damage and techniques to determine DNA damage in spermatozoa are described. A survey is presented of the correlation of sperm DNA with sperm morphology. CHAPTER 2 All the materials and methods applicable to this study are described. Sperm morphology assessment and two different sperm DNA tests, the chromomycin A3 (CMA3) staining test and the terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick-end labelling (TUNEL) assay, are discussed in detail. CHAPTER 3 Results obtained in this study are presented. Results include the prevalence of abnormal sperm DNA and association with sperm morphology, specifically in the p-pattern and g-pattern morphology groups. Further results include the correlation of sperm morphology and sperm DNA with fertilization in vitro, embryo quality and pregnancy outcome. The percentage CMA3 positive spermatozoa (abnormal DNA) and percentage TUNEL positive spermatozoa (abnormal DNA) had a significant negative association with normal sperm morphology. P-pattern and g-pattern morphology groups differed significantly from each other for both CMA3 and TUNEL. A significant positive association between CMA3 and TUNEL was observed. No association between the percentage normal sperm morphology, percentage CMA3 positive spermatozoa and percentage TUNEL positive spermatozoa and IUI pregnancy outcome was observed. A significant negative association between the percentage TUNEL positive spermatozoa and IVF/ICSI pregnancy outcome was established. The percentage CMA3 positive spermatozoa had a significant positive (unexpected) association with IVF/ICSI pregnancy outcome. There was no association between the three variables and IVF/ICSI fertilization rates. A significant positive association between the percentage normal sperm morphology and IVF/ICSI embryo quality was found. There was a significant positive association between the percentage CMA3 positive spermatozoa and IVF/ICSI embryo quality (unexpected). The percentage TUNEL positive spermatozoa and IVF/ICSI embryo quality was negatively associated. CHAPTER 4 Interpretation of the results and future perspectives are discussed. The CMA3 staining test and TUNEL assay has a limited ability to distinguish between the p-pattern and g-pattern morphology groups. P-pattern spermatozoa are more likely to possess poor chromatin packaging and show increased levels of DNA fragmentation, but some p-pattern patients also may have normal DNA and g-pattern patients abnormal DNA. It is recommended that a sperm DNA test should be implemented routinely in andrology laboratories for the clinical diagnosis of sperm DNA damage in patients.<br>AFRIKAANSE OPSOMMING: HOOFSTUK 1 'n Samevatting wat handel oor die toepassing van tradisionele semen parameters vir die evaluasie en diagnose van manlike infertiliteit, asook die rol van voorspellingswaardes in kunsmatige voortplantingstegnieke word voorgelê. Die belangrikheid van sperm morfologie, met die klem op sperm morfologie evaluering, word ook bespreek. 'n Oorsig van sperm DNS fisiologie, die proses van spermatogenese, sowel as die sperm se bydrae tot die embrio word hier aangebied. Die verskillende oorsake van sperm DNS skade en die tegnieke om sperm DNS skade vas te stel, asook die die korrelasie tussen sperm DNS en sperm morfologie word ook bespreek. HOOFSTUK 2 Alle materiale en metodes wat van toepassing is op hierdie studie word beskryf. Sperm morfologie evaluering en twee verskillende sperm DNS toetse, die chromomycin A3 (CMA3) kleuringstoets en die “terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick-end labelling (TUNEL)" toets, word ook in meer besonderhede aangebied. HOOFSTUK 3 Resultate wat verkry is tydens hierdie studie word hier uiteengesit. Resultate behels die voorkomsyfer van abnormale DNS en die assosiasie met sperm morfologie, spesifiek in die p-patroon en g-patroon. Verdere resultate sluit die korrelasie van sperm morfologie en sperm DNS met bevrugting in vitro, embriokwaliteit en swangerskap uitkomste in. Die persentasie CMA3 positiewe sperme (abnormale DNS) en persentasie TUNEL positiewe sperme (abnormale DNS) het 'n betekenisvolle negatiewe assosiasie met normale sperm morfologie getoon. P-patroon en g-patroon morfologie groepe het betekenisvol van mekaar verskil vir beide CMA3 en TUNEL. 'n Betekenisvolle positiewe assosiasie is tussen CMA3 en TUNEL waargeneem. Geen assosiasie is tussen die persentasie normale sperm morfologie, persentasie CMA3 positiewe sperme en persentasie TUNEL positiewe sperme en IUI swangerskap uitkomste waargeneem nie. 'n Betekenisvolle negatiewe assosiasie is tussen die persentasie TUNEL positiewe sperme en IVB/ICSI swangerskap uitkomste vasgestel. Die persentasie CMA3 positiewe sperme het 'n betekenisvolle positiewe (onverwags) assosiasie met IVB/ICSI swangeskap uitkomste opgewys. Daar was geen assosiasie tussen die drie veranderlikes en IVB/ICSI bevrugting nie. 'n Betekenisvolle positiewe assosiasie is tussen die persentasie normale sperm morfologie en IVB/ICSI embryo kwaliteit waargeneem. Daar was 'n betekenisvolle positiewe assosiasie tussen die persentasie CMA3 positiewe sperme en IVB/ICSI embrio kwaliteit (onverwags). Die persentasie TUNEL positiewe sperme het 'n negatiewe assosiasie met IVB/ICSI embrio kwaliteit getoon. HOOFSTUK 4 Interpretasie van die resultate en toekomstige vooruitsigte is bespreek. Die CMA3 kleuringstoets en TUNEL toets het 'n beperkte vermoë om tussen die p-patroon en g-patroon morfologie groepe te onderskei. P-patroon spermatozoa sal heel waarskynlik oor swakker chromatien verpakking en meer DNS fragmentasie beskik. Sommige p-patroon pasiënte mag egter normale DNS toon, terwyl g-patroon pasiënte abnormale DNS het. Die implementering van 'n sperm DNS toets in andrologie laboratoriums, vir die kliniese diagnose van sperm DNS skade in pasiënte, word aanbeveel.

Overall, this doctoral study aimed to contribute to the existing knowledge and clinical practice regarding male infertility, especially in relation to the potential sources and clinical impact of sperm apoptosis and reactive oxygen species (ROS) in the context of assisted reproductive technology (ART). This thesis encompasses three independent studies, each of which addresses a single aim related to different aspects of male infertility and ART practices. Study 1 examined the relationship between sperm apoptosis and ROS levels and ART outcomes. On the day of oocyte retrieval, the levels of sperm apoptosis (measured using Annexin V staining) and intracellular ROS (measured using dihydroethidium staining) were measured in neat and processed semen samples from a total of 170 male patients whose partners were undergoing in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI) treatment cycles. In general, higher levels of apoptosis and ROS were associated with lower sperm motility, sperm count and advanced paternal age. No correlation was observed between sperm apoptosis and fertilisation rate, blastulation rate, day 5 embryo quality (D5EQ) and clinical pregnancy. Similarly, there was no correlation between sperm ROS levels and fertilisation rate and clinical pregnancy. However, higher levels of ROS might be associated with poorer early stages of embryonic development, as indicated by blastulation rate and D5EQ. Together, the findings suggest that although sperm apoptosis and ROS had an adverse effect on sperm parameters, the effect on clinical outcomes was minimal. Study 2 compared the efficacy of three sperm preparation methods, namely density gradient centrifugation (DGC), the swim-up method and SeaforiaTM, in reducing the levels of sperm apoptosis and ROS. The measurement of sperm apoptosis and ROS levels were performed on a total of 17 pooled semen samples before and after sperm preparation. The main findings suggest that all three methods effectively reduced sperm apoptosis levels in the neat samples. However, only swim-up and SeaforiaTM were effective in reducing the ROS levels. Compared with DGC and swim-up, SeaforiaTM also produced samples with a higher total yield. Study 3 investigated the effects of different time intervals during ART sperm preparation on clinical outcomes. Three time intervals including pre-wash interval (time between ejaculation and sperm preparation), post-wash interval (time between sperm preparation and insemination/sperm injection) and total interval (time between ejaculation and insemination/sperm injection) were calculated and retrospectively analysed from a total of 8,079 IVF/ICSI cycles. The results showed that prolonged pre- and post-wash and total intervals had an adverse effect on sperm motility and clinical outcomes including fertilisation rate and pregnancy outcomes. Intracellular ROS production during prolonged incubation was considered to be the most likely facilitator of this observed effect. In summary, this thesis demonstrates that although their effect on ART outcomes were at low levels, higher levels of sperm apoptosis and ROS were associated with poorer sperm parameters, and suggestive of, a lower chance of natural conception. The cumulative findings of this thesis are indicative of the central role of ROS levels in sperm physiological functions and suggest that minimising intracellular ROS production through sperm preparation techniques and time optimisation may be beneficial to improving ART outcomes

Seven aerobic field trichloroethene (TCE) bioremediation projects were evaluated to determine key parameters leading to in situ TCE bioremediation effectiveness. Key parameters identified were: 1) presence of other contaminants, 2) efficacy of the cometabolic inducer, 3) technology design, and 4) site soils and hydraulics. These four parameters were then used to evaluate the pilot bioremediation operation at Air Force Plant #44 in Tucson, Arizona. The pilot operation was poorly designed. Site characterization appeared insufficient; laboratory studies were not representative of site conditions; 1, 1-dichloroethene appeared to inhibit TCE degradation; the purpose of the injected methanol ( cometabolic inducer) was unclear. Well design, specifically screen interval location, also contributed to technology deficiency. Soil type appeared to be the most limiting component; hydraulic conductivity (K) representative of the contaminated clay at the APP #44 site was estimated at 1.5 x 10-5 cm/sec. Over the course of the trial, spatially averaged TCE concentrations decreased by 41 %. Well chloride data calculations indicated that a 27% reduction may be attributable to dilution, thereby suggesting that only a 14% decrease in concentrations may be attributable to biological degradation.

O objetivo central deste trabalho é implementar modelos constitutivos hiperelásticos não lineares em um código computacional que faz análise não linear geométrica de cascas. São necessários, para este propósito, conceitos sobre álgebras linear e tensorial, cinemática, deformação, tensão, balanços, princípios variacionais, métodos numéricos e hiperelasticidade. Tal programa usa a formulação Lagrangiana posicional, o método dos elementos finitos, o princípio dos trabalhos virtuais e o método iterativo de Newton-Raphson para solução das equações não lineares. O elemento finito de casca possui dez nós, sete parâmetros por nó e variação linear da deformação ao longo da espessura. Para dedução dos novos modelos usou-se a decomposição multiplicativa do gradiente da função mudança de configuração, o tensor deformação de Green-Lagrange e o tensor da tensão de Piola-Kirchhoff de segunda espécie. O código desenvolvido foi usado em simulações de diversos exemplos e apresentou boa precisão na análise mecânica de polímeros naturais altamente deformáveis. A ocorrência do fenômeno travamento não se manifestou nas análises realizadas. A presente pesquisa confirmou outros trabalhos, reforçou a necessidade de se usar modelos hiperelásticos não lineares para simular o comportamento mecânico de polímeros naturais e apresentou resultados condizentes com dados experimentais existentes na literatura científica e às respectivas soluções analíticas.<br>The main objective of this work is to implement nonlinear hyperelastic constitutive models in a computational code of geometrically nonlinear analysis of shells. For this purpose, concepts of linear and tensor algebras, kinematics, strain, stress, balances, variational principles, numerical methods and hyperelasticity are necessary. Such program uses the positional Lagrangian formulation, the finite element method, the principle of virtual work and the iterative method of Newton-Raphson for the solution of the nonlinear equations. The shell finite element has ten nodes, seven parameters per node and presents linear variation of the strain along the thickness. To achieve the new constitutive models the multiplicative decomposition of the deformation gradient, the Green-Lagrange strain tensor and the second Piola-Kirchhoff stress tensor are used. The developed code is tested for simulations of various examples and presents good accuracy in the mechanical analysis of highly deformable natural rubber. The locking phenomena didn\'t appear in the proposed analysis. The present research confirms other works, corroborates the need of using nonlinear hyperelastic models to simulate the mechanical behavior of natural rubber and presents suitable results when compared to existent experimental data of the scientific literature and to the respective analytical solutions.

AIM OF THE THESIS Since Metabolic Syndrome (MetS) is essentially based on increased adiposity and it is associated with male hypogonadism, erectile dysfunction, psychological disturbances, and BPH/LUTS, and all these factors might, in different ways, affect reproductive capacity, we investigated their possible correlations with MetS. Hence, we performed two studies. In the first study (study 1) we evaluated possible associations between MetS, semen and hormonal parameters, as well as clinical characteristics, including sexual, ultrasound and psychological characteristics, in a cohort of men with couple infertility. In the second study (study 2), we systematically investigated the possible associations between MetS and prostate-related symptoms and signs in a cohort of young men in infertile unions and tried to establish whether these associations correlate with fertility. Study 1 conclusions We report that an increasing number of MetS factors are dose-dependently associated with relevant organic (poor sperm quality, hypogonadism, ED) and psychological (depression, somatization) features that might affect reproductive outcomes of men seeking medical care for couple infertility. This might tailor ad hoc therapeutic intervention. Behavioural interventions targeting lifestyle factors, such as dietary practice and physical activity, might ameliorate not only metabolic and psychological parameters but also male infertility, as has been demonstrated for female infertility. Study 2 conclusions This study demonstrates that in a cohort of men with infertility, a component-dependent, stepwise association was observed between an increase in the number of MetS components and the total and transitional zone prostate enlargement and prostate related-inflammatory signs but not symptoms or current infection of the male genital tract, which suggests a sub-clinical inflammation of the prostate. Relative prostate overgrowth may also correlate with MetS-related hyperinsulinaemic state. In addition, MetS but not MetS’s related prostate CDU abnormalities was associated with poor sperm morphology. FINAL CONCLUSIONS In men with couple infertility, MetS is associated with hypogonadism, poor sperm morphology, testis ultrasound inhomogeneity, erectile dysfunction, somatization and depression. In addition, MetS is positively associated with prostate enlargement, biochemical (seminal interleukin 8) and ultrasound-derived signs of prostate inflammation but not with prostate-related symptoms, which suggests that MetS is a trigger for a subclinical, early-onset form of benign prostatic hyperplasia. Recognizing MetS could help patients to improve not only fertility but also sexual and overall health.

Maintaining reproductive health and diagnosing and treating conditions of infertility in stud dogs is important in canine theriogenology. However, there is still a great deal to be learned about reproductive physiology and factors that affect reproductive organs and semen quality in dogs. This thesis is an investigation of two factors in the male dog; serum 25-hydroxy Vitamin D (25OHVD) and prolactin (PRL) concentrations, and their possible associations with benign prostatic hyperplasia (BPH), prostate volume and/or sperm morphology and motility characteristics. 28 (Vitamin D Study) and 29 (28 plus one for the Prolactin study) client dogs of various breeds from the Ontario Veterinary College and Graham Animal Hospital in Southwestern Ontario, Canada were enrolled in the study from March to December 2009. Of these dogs 22 were successfully collected for semen. BPH was diagnosed using prostate volume measured by ultrasound, as well as clinical signs including blood in the ejaculate. Semen analysis was performed using manual microscopic techniques for morphology and computer assisted sperm analysis equipment for motility. In the vitamin D study, no associations were found between BPH and serum 25OHVD concentrations. In contrast, several sperm motility (motility, progressive motility, beat cross frequency (BCF), distance average path (DAP), curvilinear distance (DCL), linear distance (DSL), average path velocity (VAP), curvilinear velocity (VCL) and straight line velocity (VSL), amplitude lateral head displacement (ALH) and average orientation change (AOC)) and morphology characteristics (percentage normal sperm, head defects and detached heads) had desirable outcomes with 25OHVD concentrations between 120-180 nmol/l. Using bivariable analysis, positive associations were observed with 25OHVD and some semen quality characteristics from 4 to 8 years of age (motility, progressive motility, BCF, DCL, VCL, ALH, AOC) and at transformed prostate volumes smaller than or equal to 4.5 (motility, progressive motility, DCL, VCL, and normal morphology) while negative associations of these semen parameters were found at ages greater than 8 years and transformed prostate volumes greater than or equal to 5.5. Head defects were negatively associated with 25OHVD. Vitamin D may have an impact on spermatogenesis and normal sperm physiology that warrants further research. The prolactin study showed no statistically significant associations between serum PRL and BPH and serum PRL and sperm motility characteristics. However, two sperm morphology characteristics (percentage proximal droplets and percentage midpiece defects) had significant negative associations with PRL concentrations. Age interaction with PRL was also a factor in the percentage of midpiece defects with desirable outcomes associated at 4 years of age compared with older ages. Overall, undesirable outcomes occurred at PRL concentrations less than 2.5 ng/ml. In conclusion, both 25OHVD and PRL may have important roles in spermatogenesis and normal sperm physiology in the dog.<br>Ontario Veterinary College Pet Trust

The presence of pathogens in semen can compromise the outcome of assisted reproductive treatment, together with the possibility of the female partner or offspring becoming infected. This is cause for concern, especially in South Africa with a high prevalence of HIV-1. Most of these infected individuals are in their reproductive years with the desire to have their own genetically related children. Therefore, assisted reproductive treatment with effective risk reduction procedures, such as semen processing for the elimination of these pathogens is crucial. However, during sperm preparation by standard discontinuous density gradient centrifugation, the supernatant is aspirated to allow access to the purified sperm pellet. Pathogens from the upper layers can adhere to the inside surface of the test tube and flow down to re-infect the purified sperm sample. The use of a centrifuge tube insert may prevent the re-contamination of sperm samples after discontinuous density gradient centrifugation. Furthermore, seminal pathogens can bind specifically or non-specifically to spermatozoa, rendering semen decontamination procedures ineffective. Serine proteases, such as trypsin, have been demonstrated to effectively inactivate viruses and to break pathogen-sperm bonds. However, the addition of a protease to density gradient layers during semen processing could have a negative impact on sperm parameters. This research was therefore aimed towards the determination of: i) The effect of semen processing with trypsin and trypsin inhibitor on sperm parameters. ii) The prevalence of various bacteria in semen samples from men attending the Reproductive and Endocrine Unit at Steve Biko Academic Hospital. iii) The effectiveness of semen processing by discontinuous density gradient centrifugation with a centrifuge tube insert, for the elimination of some of the most prevalent bacteria, white blood cells and in vivo derived HIV-1. Evaluation of sperm parameters after semen processing indicated that trypsin and trypsin inhibitor did not have an impact on sperm mitochondrial membrane potential, vitality, motility and zona binding potential, or acrosin activity, respectively. Seminal bacteria were highly prevalent in patients wishing to participate in the Unit’s assisted reproductive program, with 49.5% of semen samples presenting with positive bacterial cultures. Semen processing by means of discontinuous density gradient centrifugation with the tube insert, eliminated significantly more in vitro derived (spiked) bacteria and white blood cells from semen compared to processing without the insert. Furthermore, the semen decontamination procedure was effective in removing HIV-1 RNA from 100% of samples and proviral DNA from 98.1% of semen samples from HIV-1 sero-positive patients. The effectiveness of discontinuous density gradient centrifugation for the elimination of seminal pathogens could, therefore, be improved by the addition of trypsin to the upper density layer, without supplementing the bottom layer with trypsin inhibitor. Additionally, semen decontamination efficiency could also be improved by the prevention of re-contamination of processed sperm samples by the utilization of a tube insert during discontinuous density gradient centrifugation.<br>Thesis (PhD)--University of Pretoria, 2014.<br>gm2014<br>Obstetrics and Gynaecology<br>unrestricted

The current generation of cosmological hydrodynamical simulations offer new levels of fidelity in galaxy formation and evolution that can be benchmarked against observations. However, it is crucial that the comparison between the simulation products and observations is performed on level-ground. Using mock-observations of galaxies from hydrodynamical simulations with observational realism, an image-based comparison is enabled between the simulations and modern galaxy surveys that allow galaxy properties to be derived consistently. A new methodology is presented that provides an unprecedentedly comprehensive suite of observational realism to synthetic images of galaxies from simulations and performs detailed decomposition of their morphological structures. The crux of the methodology is that the same procedure for image-based surface-brightness decompositions of galaxy structures is employed for the simulations and observations -- facilitating a fair and unbiased comparison of galaxy properties. The methodology is piloted on galaxies from the Illustris simulation and is designed to enable comparison with galaxies from the Sloan Digital Sky Survery (SDSS). The biases from observational realism on the decomposition results for the simulated galaxies are characterized in detail using several controlled experiments. Then, the decompositions are used in a comparison of the galaxy size-luminosity and bulge-to-total vs. total stellar mass relations. The comparisons show that galaxies from Illustris contain too many discs and too few bulges at low masses M*/M☉≤11 relative to the SDSS. A comparison of the photometric and kinematic bulge-to-total is also enabled by the methodology. The comparison suggests that photometry tends to systematically under-estimate the bulge fractions relative to the kinematics -- allowing no discernible connection to be made between photo-bulges and kinematic bulges. Several possibilities that may be driving the bulge deficit in Illustris' low-mass galaxies are discussed, though particle resolution is argued to be the main culprit. The methodology that is presented in this thesis has broad applications to comparisons between computational and observational galaxy astronomy and stands to provide a wealth of feedback between each community.<br>Graduate<br>0606<br>cbottrel@uvic.ca

碩士<br>國立中興大學<br>生命科學院碩士在職專班<br>95<br>The semen analysis is most often used for evaluation of the seminal fluid quality and its fertility potential. The traditional semen analysis is subjective and its result has high variations. The computer-assisted sperm analyzer (CASA) may provide an objective, precise, and relative high reproducible value on quantification of sperm motility characteristics. The purpose of this study is to determine the relationship between sperm characteristics, including motility and morphology, and in vitro fertilization rates. In this study, the CASA execution semen analysis was used, the seminal fluid test samples came from sterile husbands and wives who were on the treatment course of in vitro fertilization are in total of 66 couples. The CASA analysis was used to compare the fresh seminal fluid and PureSperm-screened sperm for motility characteristics and fertilization rates of in vitro test samples. The sperm morphology is evaluated by the light microscope according to Kruger’s strict criteria. The findings showed that, for the fresh seminal fluid, the progressive motile sperm proportion, straight line velocity (VSL), the linearity (LIN), and sperm morphology have a remarkable relevance with the rate of fertilization. However, for those PureSperm screened sperm''s test samples, only the percentage of progressive motile sperm (r = 0.27, p<0.05) is significantly correlated with fertilization rates. Our results confirmed that the factors including progressive motile proportion, VSL, LIN, and normal form of sperm can be used as prognostic indicators for evaluation of fertilization potential of fresh seminal fluid. We found that the progressive motile sperm proportion is the only factor which can be used to forecast fertilization potential on PureSperm screening sperm''s test sample. The above results will be useful to aid the laboratory in planning its strategy on insemination and to reduce probability of fertilization failure.

Aim of this thesis was to evaluate influence of inorganic selenium given, in a form of mineral supplement compound, to cows and their calves in order to supply their organism with selenium and also for the utilitarian traits. These were divided into two groups of six and supplemented with two different forms of selenium: group A was given inorganic sodium selenite, group B was given organic selenomethionine. Cows that were examined before the start of supplementation had average activity GPX in blood samples from group A 847,06 +- 170,77 ukat/l and group B 791,30 +- 91,20 ukat/l and concentration of Se of group A: 136,70 +- 27,97 ug/l and group B 95,77 +- 20,05 ug/l. It was found that cows postpartum had higher activity of GPx by 26% in group A and by 45% in group B. Difference between groups was statistically inconclusive (p < 0,28). Increase in concentration of selenium in blood of cows in group A was 7,5% and in group B 5%. In blood of newborn calves of both groups was not found conclusive difference of GPx (p < 0,43). Concentration of selenium in group calves A 160,18 +- 18,07 ug/l and group B 105,30 +- 24,26 ug/l. Difference in concentration of selenium in blood of cows and calves was statistically conclusive (p < 0,05), but because this difference was conclusive already before the supplement compound was given both inorganically and organically bound selenium, this result cannot be interpreted as conclusive influence of different forms of selenium supplemented. Weight and weight gain of calves were balanced between groups and statistically did not differ (p < 0,39). It can be concluded that both forms of selenium had analogical biological effect on focused parameters.In blood of newborn calves no conclusive difference in GPx was found

The aim of this master thesis was to experimentally determine the effect of grape seeds (their metabolically active compounds) on blood parameters of laboratory rats. The experiment was conducted in a laboratory of The Mendel University, Brno in late 2015. Four groups of laboratory rats were supplied with; 1) water, 2) water in combination with grape seed extract, 3) water with grape seed extract and cadmium, 4) water in combination with cadmium. Half of the laboratory animals were killed after 14 days and second half were killed after 28 days. Blood samples were analyzed in the veterinary laboratory (VLAB). The results were evaluated and compared. From our results, we can assume that feeding grape seed extract for its protective effect against damage of organism caused by cadmium, cannot be recommended because this effect was not proved.

Introduction. Sperm parameter analysis is the standard method of male fecundity testing; however, minimal evidence supports associations between individual sperm parameters and reproductive outcomes. Our previous work shows strong associations between sperm mitochondrial DNA copy number (mtDNAcn) and time-to-pregnancy (TTP) in general populations, and between mtDNAcn and fertilization outcomes in clinical populations. Thus it is possible for sperm mtDNA biomarkers to act as summary measures of semen quality. In this study, we developed a sperm quality index (SQI) from semen parameters and compared its ability to measure fecundity to sperm mtDNAcn. Methods. We received 384 semen samples from the Longitudinal Investigation of Fertility in the Environment Study. Sperm mtDNAcn and mtDNA deletions (mtDNAdel) were quantified using a triplex probe-based qPCR method. The SQI was developed by ranking and summing select sperm parameters within the study population, including sperm concentration, sperm count, normal morphology, high DNA stainability, and DNA fragmentation to create a cumulative index. Discrete-time proportional hazards models were used to determine fecundability odds ratios (FOR), indicating associations between mtDNAcn, SQI, and TTP. Receiver operating characteristic (ROC) analyses determined the validity of the SQI and mtDNAcn as predictors of pregnancy within 12 months. Results. The SQI was highly associated with mtDNAcn, both continuously (Spearman Rho: -0.487; p-value: <0.001) and in deciles (ANOVA p-value: <0.001). The SQI (FOR: 1.25; 95% confidence interval (CI): 1.09, 1.43) and mtDNAcn (FOR: 0.754; 95% CI: 0.657, 0.866) performed similarly in discrete-time survival models and indicated a significant decrease and increase in TTP, respectively. MtDNAcn more effectively predicted pregnancy within 12 months (AUC: 0.703; 95% CI: 0.617, 0.789) than the SQI (AUC: 0.642; 95% CI: 0.531, 0.753). With multiple predictors, mtDNAcn outperformed summary models, with addition of the SQI and percent normal morphology minimally increasing model efficacy (AUC: 0.718, 95% CI: 0.617, 0.819). Conclusion. The association between the SQI and mtDNAcn suggest that mtDNAcn may serve as a summary biomarker for overall sperm quality. Neither individual nor summed sperm parameters are useful indicators of couple fecundity and reproductive outcomes compared to mtDNAcn. These results suggest that mtDNAcn has potential for use as a biomarker of fecundity.

The theoretical part is concerned with the functional application of selenium and iodine supplementation methods, risks disproportionate to their income, including changes in biochemical and hematological parameters and evaluation methods of adequate saturation. The practical part analyzes the results of experiments on sheep with a defined content of selenium and iodine in the ration evaluation of selected indicators of the metabolic profile of ewes and their lambs. The results are processed statistically. It reflected the dynamics and interactions of monitored parameters.

The aim of this thesis was to evaluate the quantitative and qualitative parameters of boar semen for insemination center Radouňka. There were 74 breeding boars of Czech Large White, Czech Landrace and line L48 monitored at the station. Basic quantitative and qualitative parameters of semen were monitored (volume in ml, sperm concentration in 1mm3. motility in % and the percentage of pathological sperms) which were assessed according to breed, age of boar, influence of season and sperm production. The individual parameters of breeding boar?s semen were monitored from January 2009 to December 2011.