Dissertations / Theses on the topic 'Semen quality'

Artificial insemination (AI) is a widely used part of the modern agricultural industry, with the number of animals inseminated globally being measured in the millions per anum. Crucial to the success of AI is that the sperm sample used is of a high Quality. Two factors which determine the quality of the sample are the number of sperm present and their motility. There are numerous methods used to analyse the quality of a sperm sample, but these are generally laboratory based, expensive and in need of a skilled operator to perform the analysis. It would, therefore be useful to have a simple and inexpensive system which could be used outside the laboratory, immediately prior to the insemination of the animal. Presented in this thesis is work developing a time of flight (ToF) technique which makes use of a quartz crystal microbalance (QCM), operating at 5 MHz, as the sensing element. Data is shown developing a device where a 50 μl sample of boar sperm is added to a liquid filled swim channel, which the sperm are allowed to self-propel down and attach to the surface of a QCM at the end. The attachment of the sperm to the surface causes a measurable frequency decrease in the QCM, aproximately 50 Hz. An average effective mass measurement was made using a QCM and gave a value of 8 ± 5 pg per sperm, which was used in conjunction with the frequency change to determine the number rate of sperm reaching the QCM. Additional data is presented to investigate the effect of environmental temperature on the ToF of the sperm, showing a decrease in ToF between 23 0C to 37 0C. The system was also used to investigate increasing the swim speed of the sperm by chemical means. A range of 20 μmol to 100 μmol of progesterone was added to the swim medium and the ToF was shown to decrease as a result. To further develop the system, large commercial electronics were replaced by smaller circuits built in-house. An oscillator circuit based on a Pierce oscillator was used to drive the QCM and a frequency counter circuit making use of a universal frequency to digital converter (UFDC-1) was used to measure the frequency of the QCM. ToF experiments were performed which showed these pieces of equipment to be effective for performing the analysis of sperm samples. The swim cell itself was also refined, resulting in a compact, modular design. Work was performed developing layer-guided, single-port acoustic resonators to replace the QCM as the sensing element in the sperm analysis device. A maximum mass sensitivity of 1110 Hzμg-1cm-2 was found for devices on a LiTaO3 substrate with a 6 μm guiding layer. While viscosity-density sensing experiments found a maximum sensitivity of 488 KHz Pa-1/2 kg1/2 for a 4 μm guiding layer.

POOLPERM, PARIWAT.  Factors Influencing Semen Quality and Fertilityin Boars. (Under the direction of Drs.Glen W. Almond and William L. Flowers)

        The objectives of this researchwere 1) determine the influence of antibiotics on semen quality, 2) determinethe association among insulin-like growth factor I (IGF-I) in seminal plasma,semen quality, and subsequent fertility, and 3) retrospectively study theassociation among semen parameters and sow fertility.  In the firststudy, the effects of gentamicin (GM), amikacin (AM), neomycin sulfate(NM), and penicillin-streptomycin (PS) in semen extender on the percentagesof motile (MOT), morphologically normal sperm (MOR), and sperm with normalacrosome (NAR) were examined.  An in vitro penetration assay was conductedusing sperm cells on day 0 and day 5 of storage.  GM and NM groupsshowed higher (p<0.05) MOT than other groups after 5 days of storage. No differences in penetration rate were found among treatments; however,the penetration rate decreased (p<0.05) on day 5 of storage.
        In the second study, ejaculateswere collected and diluted in an extender (Vital?).  Gilts (n=113)and sows (n=375) were inseminated with the extended semen in homogenetic-homospermicregimens.  Farrowing rate (FR), total pigs born (TB) and born alive(TBA) were recorded.  IGF-I was determined in seminal plasma by radioimmunoassay. Concentration of IGF-I from 204 ejaculates was 95.38 ± 3.56 ng/ml (mean± SEM) and total amount of IGF-I/ejaculate was 23.50 ± 1.20 µg.  SeminalIGF-I differed (p<0.05) among genetic lines and had no effect (p>0.05)on MOT, MOR and NAR.  However, IGF-I was associated (p<0.05) withsemen volume, sperm concentration and total number of sperm/ejaculate. No association between IGF-I level in seminal plasma and fertility indiceswas found.
        The third study determinedthe associations among insemination parameters with subsequent fertilityof boar semen using data from the second study.  MOR was associated(p<0.05) with fertility parameters.  TB and TBA were associatedwith age of semen at the first insemination (SAGE) and number of spermper insemination dose (AIDOSE).  With stepwise regression analysis,it was evident that FR was associated with semen volume, MOR and SAGE. Meanwhile, TB and TBA were associated with SAGE, AIDOSE and total numberof spermatozoa/ejaculate.  In conclusion, the assessment of semencharacteristics may not necessarily delineate fertility between boars.

The primary objective of this research was to study the effects of breed, age, season, and their interactions on semen morphological characteristics. The study was done on 329 bulls (271 Friesland and 58 Jersey) aged 12, 24, 36,48, 60, 72, 84, 96 and> 96 months. The collection of semen was carried out using the artificial vagina method in all four seasons of the year. Spermatozoa were screened for the percentages normal sperm, percentage and total major defects such as knobbed acrosome, pyriform, abnormal lose head, dag defects, nuclear vacuole, degenerative heads, mid-piece reflexes, percentages and total minor defects such as normal lose heads, distal droplets, curled end-piece, lose acrosome. Statistical analyses of the data were done using the general linear model (GLM) procedure of the Statistical Analyses System (SAS, 1999). The results of the study indicate that breed did not significantly affected the percentage normal sperm and percentage major sperm defects, but significantly affected the percentage minor defects (P = 0.01). The Least square means (LSM±SE) for the percentage normal sperm, major defects and minor defects in Friesland and Jersey bulls were 80.6 ±1.06%; versus 78.9±2.31 %; 14.8±0.90% versus 15.0± 2.62%, 5.1±0.43% versus 7.6±0.94%, respectively. The results obtained show that the prevalence of sperm defects that differed significantly between breeds was higher in Jersey bulls compared to Friesland bulls. The results of the study indicated the percentage of normal sperm to differ (P = 0.01) with season. The percentage of normal sperm during the summer, autumn, winter and spring, were 72.8±1.6%, 79.4±2.2%, 82.5±2.4% and 84.4±2.4% respectively. Season also affected the percentage of major defects (P = 0.01) and percentage of minor defects (P = 0.03). The results demonstrate that even though there was a higher variation in sperm morphology with season, better sperm morphology was recorded in spring and winter than summer and autumn. Results also indicate the percentage of normal sperm (P = 0.05) and major defects (P = 0.01) to be affected significantly by age. On the other hand, the percentage of minor defects did not differ significantly with age. Bulls of 36-48 months of age showed better semen quality than bulls older than 72 months and bulls younger than 36 months. The percentage of major defects, particularly the incidence of major defects such as knobbed acrosomes, pyriforms, dag defects and broken flagella were significantly affected by the interaction between age and breed (P = 0.05) and age and season (P = 0.05). There was an increase in the susceptibility to these sperm defects in Jersey bulls with an increase in age, while no variation was observed in Friesland bulls. With age and season combined, young bulls recorded poor semen morphology during winter, while old bulls showed poor morphology during summer. In conclusion, the study suggested that breed, age and season and their interactions are important sources of variation in sperm morphology. For a successful AI programme, semen collection should be done at the age of 36-48 months for both breeds. It is therefore recommended that age, breed and season should be given urgent attention in any bull management system employed in South Africa in order to obtain the best semen quality.
Dissertation (M Inst Agrar (Animal Production))--University of Pretoria, 2003.
Animal and Wildlife Sciences
unrestricted

The chromosomal location of endocrine genes was established, and relationships between expression of specific endocrine genes and measures of testis function in normal and poor semen quality stallions was assessed. Consensus primer sequences for glucocorticoid receptor (GR) and luteinizing hormone receptor (LHR) were used to screen the CHORI-241 equine bacterial artificial chromosome (BAC) library. The identity of PCR-positive BAC clones was confirmed by sequencing. Verified BACs were mapped to horse metaphase chromosome spreads by fluorescence in situ hybridization (FISH). The BACs containing the GR and LHR were localized by FISH to ECA 14q16-q21 and ECA15q22-q23, respectively. In addition to FISH mapping, the 5000rad horse x hamster radiation hybrid (RH) panel was screened in duplicate. Two-point linkage analysis placed GR 0 cR from LEX047, while LHR was 36.67 cR from TKY011 on ECA14 and ECA15, respectively. Total testicular parenchymal weight, mean daily sperm production (DSP) per gram parenchyma and mean apoptotic rate (406.05 ± 24.33g vs. 180.01 ± 34.41g, 15.29 ± 0.87 vs. 10.24 ± 1.10, 6.70 ± 0.88 vs. 14.25 ± 1.11, respectively) differed (P<0.05) between normal (n=8) and poor semen quality (n=5) stallions. Also, plasma estradiol and inhibin concentrations were higher (P<0.05) in normal stallions than in poor semen quality stallions. Testicular expression of estrogen receptor beta (ER beta), βB inhibin, prolactin receptor (PRLR), growth hormone receptor (GHR) and insulin-like growth factor I receptor (IGF-IR) mRNAs were all lower (P<0.05) in poor semen quality stallions than in normal stallions. The BACs and primers developed in this study will facilitate future investigations of GR and LHR gene structure in the horse as well as providing a resource for physiological investigation of these two genes that are primary regulators of stress responsiveness and fertility. These data add important endocrine genes to the horse cytogenetic map. Also, important hormonal and gene expression changes have been identified in poor semen quality stallions for further investigation.

Centrifugation of stallion semen is an integral part of the cryopreservation procedure, primarily allowing for the concentration of sperm and removal of seminal plasma. In addition, centrifugation is required for maximizing spermatozoal quality in semen from some stallions subjected to cooled transport, because of the detrimental effects of long-term exposure to high levels of seminal plasma. The centrifugation process, however, has potential deleterious effects, including reduction in sperm quality as well as loss of sperm numbers. Since centrifugation plays such a crucial role in semen processing, two experiments were designed to evaluate more efficient centrifugation methods to meet the demands of the equine industry. In Experiment 1, semen was centrifuged in two different tube types (nipple- or conical-bottom), using a cushioned technique (Eqcellsire® Component B) with two different extenders (opaque-INRA96 or clear-HGLL). For Experiment 2, nipple-tube centrifugation was conducted at two different g forces (400 or 600) for 20 min, using three different iodixanol cushion media, Eqcellsire® Component B, OptiPrep[TM], or Cushion Fluid[TM]. Regardless of tube or extender types, centrifugation of semen resulted in sperm recovery rates ≥90%; however, centrifugation in INRA 96 extender yielded higher sperm motility values than did centrifugation in HGLL extender (P < 0.05). Cushion type or g force did not impact post-centrifugation semen quality, based on the laboratory values measured (P > 0.05). These results indicate that cushioned centrifugation of stallion semen in either conical-bottom or nipple-bottom tubes can yield a high sperm harvest, while maintaining sperm function. An optically opaque extender, as is typically used in the equine breeding industry, can be used to achieve this goal. The fertility rate (94%; 131/140) following cushioned semen centrifugation in a commercial program this past year indicates that these laboratory results are transferable to the clinical setting.

The aim of this study was to determine the efficacy of vaccination in preventing LSDV excretion in semen and negative effects on semen quality. Lumpy skin disease (LSD) is caused by a virus in the genus Capripoxvirus of the family Poxviridae. The virus has been reported to be excreted in the semen of experimental infected nonvaccinated bulls. Nevertheless, vaccination has been the most widely used method to reduce and prevent the spread of the disease. This work was done to determine the efficacy of lumpy skin disease vaccination in preventing the excretion of lumpy skin disease virus (LSDV) in semen of experimentally infected vaccinated bulls. It also determined further the effect of vaccination and experimental infection on semen quality. Six serologically negative bulls 11-16 months of age were vaccinated with an attenuated Neethling strain of LSD vaccine, and a repeated dose of vaccine was given twenty one days later. These bulls were then experimentally infected by intravenous injection with a virulent field strain of LSDV (V248/93). Six unvaccinated bulls were similarly infected to act as controls. All animals were observed for clinical signs, blood and semen was collected and evaluated twice a week until day 40 post vaccination and every two days until day 28 post-infection when the trial was terminated. Serology was done using the serum neutralization test and viraemia was determined by virus isolation. Semen was examined by polymerase chain reaction (PCR) for the presence of virus. Semen evaluation was done visually and microscopically. Two of the unvaccinated controls developed severe LSD, two showed mild symptoms and two were asymptomatic. No clinical abnormalities were detected following vaccination, and clinical signs were limited to mild lymph node enlargement in four bulls following challenge of the vaccinated bulls. There was a significant difference (P<0.05) in semen quality after experimental infection of the unvaccinated bulls. In the vaccinated bulls, semen quality showed no significant difference (P>0.05) following vaccination and challenge. Three of the vaccinated bulls were serologically positive at the time of experimental infection and four at the end of the trial. Five unvaccinated bulls were found to be viraemic during the course of the trial. No vaccinated bulls were found to be viraemic at any stage. Four unvaccinated bulls excreted the virus in their semen during the course of the trial. Viral nucleic acid was not detected in any semen samples following vaccination or challenge in vaccinated bulls. This study provides evidence that vaccination against LSD prevented the excretion of viral particles in semen. It also illustrated that LSD vaccination prevented any effect on semen quality after experimental infection with virulent virus.
Dissertation (MSc (Production Animal Studies))--University of Pretoria, 2006.
Production Animal Studies
unrestricted

Studies have discovered higher frequencies of single nucleotide polymorphisms (SNPs) in different mitochondrial genes are associated with subnormozoospermia. However, the frequencies of SNPs in ND1 and ND2 are not unknown. The present research was aimed to determine the frequencies of SNPs in ND1 and ND2 genes of the mitochondrial genome in fertile and subfertile men and whether changes in codon usage was associated with fertility phenotypes. Total genomic DNA from 157 semen samples was extracted using the proteinase K/SDS digestion procedure, followed by phenol/chloroform purification and ethanol precipitation. ND1 and ND2 genes were amplified respectively from 80 and 92 DNA samples from different fertility groups. Each PCR product was sequenced to identify mutations. Codon change resulting from a nucleotide substitution was determined by comparison with a reference mtDNA sequence obtained from the NCBI database. The frequency of codon usage in the reference mtDNA was determined by the computer program MEGA version 2.1. Eleven synonymous nucleotide substitutions and two non-synonymous substitutions were found in this study. Four SNPs were previously characterized; all SNPs were homoplasmic. None of the SNPs were likely to affect the function of the proteins on the basis of the hydrophobicity plots or secondary structure predictions. Sixty two percent of synonymous mutations were found to change from a high to a low relative codon usage values; 37% of synonymous mutations changed from a low to a high relative usage value. Chi-square (χ²) test (χ²= 0.067 with 1 d.f.) showed that there was no significant difference at the 5% level between these changes. Thus, change in codon usage was not related to semen quality in men. Further, there were no statistically significant differences in the observed frequencies of SNPs of fertile and subfertile men. However, the sample size was small and this study was only focused on a single NZ Caucasian population. Further study including larger and more diverse population samples may provide further insight into the functional importance of codon usage and its relevance to fertility

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
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.
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.

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 in vitro techniques have yet to be established. The 'swim-up' test is suggested as the most suitable on farm in vitro semen assessment, and that the 'Anti-shock' dilution technique has commercial promise. Fructose should be the usual sugar used in the diluent.

Rift Valley fever (RVF) is an arthropod-borne viral disease of significant importance in both livestock and humans. Epidemics occur periodically in domestic ruminants, typically after heavy rains, which encourage rapid multiplication of mosquito vectors. Clinical symptoms in livestock vary from inapparent infection to abortions and peracute deaths. The disease has significant zoonotic potential. People in contact with infected livestock may develop disease that varies from mild flu-like symptoms to severe neurological and haemorrhagic disorders and death. An important way of controlling the disease is through vaccination of susceptible livestock. Rift Valley fever virus (RVFV) clone 13 is a relatively new livestock vaccine against RVF that is derived from an avirulent natural mutant strain of RVFV. This vaccine has been shown in previous studies to confer protective immunity against infection with live virus. The effect of this vaccine on semen quality in male animals has never been tested. The purpose of the current trial was to determine whether RVFV clone 13 vaccine had any effect on semen quality in rams. The hypothesis tested was that animals vaccinated with RVFV clone 13 vaccine would not experience a reduction in semen quality (measured by evaluating the percentage progressively motile and percentage morphologically normal spermatozoa in successive ejaculates) relative to unvaccinated control animals. A test/control model was used to evaluate the effect of this vaccine on semen quality. A group of peripubertal ram lambs were tested for antibodies to RVFV using a serum neutralisation test (SNT). Animals without detectable antibodies (n=23) were then randomly allocated to either a test group (n=12) or a control group (n=11). Daily rectal temperature measurements were taken and weekly semen evaluations were conducted. Blood samples were drawn weekly to assess serum antibody titres. Seven animals were subsequently eliminated from the statistical analysis because of potential confounding factors. Of these seven, five animals had extremely poor semen quality at the start of the trial, one animal was found to have a persistent febrile response commencing at the start of the trial, and one animal had seroconverted to Rift Valley fever virus in the period between the initial screening and onset of the trial. Logistic regression analysis was performed on data gathered from the remaining animals to determine whether an association existed between animal group, rectal temperature and semen quality parameters. It was found that no correlation existed between treatment group and values obtained for the semen quality parameters measured. There was no statistically significant post-vaccination temporal decline in the percentage of live morphologically normal spermatozoa, or the percentage of progressively motile spermatozoa, either when assessed amongst all animals or when assessed within individual groups. Based on the data from this trial, the hypothesis was not rejected. Despite this finding, it should be stated that the elimination of animals from the analysis had some effect on the statistical power of the study. A repeat of the trial with a larger sample size and a more comprehensive pre-screening process to avoid the inclusion of animals with poor semen quality may be indicated.
Dissertation (MMedVet)--University of Pretoria, 2014.
tm2015
Production Animal Studies
MMedVet
Unrestricted

La infertilidad en el Perú, actualmente no se encuentra considerada como una enfermedad importante para el ámbito de la salud pública, no obstante, afecta al 15% de la población en edad reproductiva y la demanda de Centros de Reproducción Asistida cada vez aumenta. Se considera que el factor masculino contribuye con el 40% de los casos como factor de infertilidad en la pareja. El análisis clínico del semen aporta al conocimiento de la calidad seminal que pueda presentar, que luego permitirá tratar a los pacientes exitosamente. El objetivo de este estudio fue evaluar la calidad seminal de pacientes con problemas de fertilidad que asistieron al Centro de Reproducción Humana de Lima (NACER) durante el periodo de Mayo del 2015 hasta Mayo del 2016. La población de estudio estuvo conformada por 150 individuos con edades comprendidas entre 27 y 70 años, a quienes se les realizó un análisis seminal, donde se evaluó parámetros seminales como volumen, viscosidad, pH, vitalidad, concentración y morfología espermática, según el manual del líquido seminal de la Organización Mundial de la Salud en su 5ta edición 2010. Se calcularon los estadísticos descriptivos, frecuencias y medias de las anomalías más frecuentes y se compararon con una condición normal en los parámetros seminales más importantes en fertilidad. Todos los análisis se realizaron con un nivel de confianza de 95% utilizando el programa SPSS versión 23 para Windows 2010, además se realizó una tabla de frecuencias que fue utilizada para obtener gráficos en excel indicando la prevalencia e incidencias de las alteraciones en los espermatogramas realizados. Del total de muestras analizadas el 50.6% presentaron por lo menos un parámetro alterado, mientras que el 49.3% de los casos están sobre los valores de referencia establecidos en la 5ta edición del manual de la OMS (2010). La astenozoospermia fue la anomalía más prevalente en los pacientes con un porcentaje de 28%, seguido de la oligozoospermia e hipospermia con un porcentaje de 19.3% en ambos casos. En las características macroscópicas se encontró que 45 (30%) de los casos evaluados presentaron anormalidades en su consistencia y el 8.6% de los casos estudiados presentaron un cuadro de azoospermia. La presencia de alteraciones como astenozoospermia, oligozoospermia, hipospermia y teratozoospermia refleja la mala calidad seminal poblacional que viene experimentando el hombre en los últimos años.Infertility in Peru is not currently considered an important disease for the field of public health; however, it affects 15% of the reproductive-age population and demand for Assisted Reproduction Centers each time increases. It is considered that the male factor contributes 40% of cases as a factor of infertility in couples. The clinical analysis of semen provides the knowledge of seminal quality that can present, that then it will allow treat patients successfully. The aim of this study was evaluate the semen quality of patients with fertility problems who attended to the Human Reproduction Center of Lima (NACER) during the period May 2015 to May 2016. The study population consisted of 150 individuals with aged between 27 and 70 years who underwent a seminal analysis, where seminal parameters was assessed such as volume, viscosity, pH, vitality, concentration and sperm morphology according to criteria of the World Health Organization (2010). Descriptive statistics, frequencies and media of the most frequent anomalies were calculated and compared with a normal condition in the most important seminal parameters in fertility. All analyzes were performed with a confidence level of 95% using SPSS version 23 for Windows 2010, also it was performed a frequency table that was used for get graphics in excel indicating the prevalence and incidence of alterations in semen analysis made. Of the total samples analyzed 50.6% showed at least one parameter altered, while 49.3% of cases fulfill the benchmarks set out in the 5th edition of the WHO manual (2010). The astenozoospermia was the anomaly most prevalent in the patients studied with a percentage of 28%, followed by oligozoospermia and hipospermia with a percentage of 19.3% each anomaly. In the macroscopic characteristics it found that 45 (30%) of cases evaluated presented abnormalities in consistency and 29% of the cases studied presented azoospermia. The presence of alterations as astenozoospermia, oligozoospermia, hipospermia and Teratozoospermia reflects the decline in semen quality population that man has experienced in recent years.

No Brasil, assim como em outros países tropicais, a criação de bovinos é feita de maneira extensiva e, portanto é suscetível às intempéries climáticas. Esse tipo de criação pode levar ao estresse térmico, mesmo em animais zebuínos. Sob condições de estresse térmico é possível a ocorrência de degeneração testicular, e por isso, há a procura por animais tolerantes ao calor. Desta maneira, este trabalho objetivou a avaliação da tolerância ao calor de touros da raça Brahman utilizando o teste de tolerância ao calor, e também a análise da qualidade seminal relacionada à disponibilidade de sombra nos pastos. Para tal, foram utilizados 10 touros da raça Brahman com idades entre 24 e 30 meses. Duas semanas antes do início das análises, três amostras de sêmen foram colhidas de cada animal para o nivelamento biológico do sêmen, e então os touros foram separados em dois grupos: 5 touros foram alocados em pasto com disponibilidade de sombra enquanto 5 touros foram alocados em pasto sem qualquer tipo de sombra. O teste de tolerância ao calor foi realizado em três dias consecutivos típicos de verão, e as colheitas de sêmen foram realizadas em 14 dias durante 2 meses, totalizando 4 colheitas. Anteriormente a todas as colheitas foram avaliadas a consistência testicular e o perímetro escrotal. As características seminais avaliadas foram volume, aspecto, turbilhonamento, motilidade, vigor e concentração e morfologia espermáticas. Os dados obtidos foram analisados com o pacote estatístico Statistical Analysis System (SAS Institute Inc., 2004). Não houve diferenças estatísticas entre os dois grupos de animais. O resultado do teste de tolerância ao calor mostrou que os animais testados são tolerantes ao calor e apresentam boa capacidade termolítica.
In Brazil, similarly to others tropical countries, bovines are bred under environmental conditions of extensive system. This breed system type may lead to heat stress even on zebu animals. Under heat stress conditions, testicular degeneration can occur and, therefore, there is a search for animals that are heat tolerant. Therefore, the aim of this work was to evaluate the heat tolerance of Brahman bulls using the heat tolerance test and to verify the occurrence of any possible effect of shadow availability on pasture on semen quality. Ten Brahman bulls aging between 24 and 30 months were used in this work. For biological semen evaluation, three semen samples were collected from each animal two weeks before beginning. The bulls were then separated into two groups: 5 animals were allocated on pasture with shadow availability and 5 bulls were allocated on pasture without any kind of shadow. The heat tolerance test were performed in three consecutive typical summer days and the semen samples were collected each 14 days during 2 months, in a total of 4 semen samples per animal. Testicular consistence and scrotal circumference were measured just before every semen collection. The semen\'s characteristics evaluated were volume, aspect, mass movement, motility, straight movement, sperm concentration and morphological exam. Data obtained from experimental proceedings were analyzed by Statistical Analysis System program (SAS Institute Inc., 2004). No difference was observed (P>0.05) in any of the characteristics analyzed when Brahman bulls were maintained on pastures with or without shadow availability. The performance of tested animals on the heat tolerance test shows that these animals are heat tolerant and present good thermolitic ability.

Veštačko osemenjavanje (VO) je najznačajnija reproduktivna biotehnologija u
intenzivnoj proizvodnji svinja. Efikasan izbor visoko fertilnih genetski superiornih
nerastova i visok fertilitet veštački osemenjenih krmača, ima veliki ekonomski uticaj na
efikasnost praktične primene ove biotehnologije. Međutim, prethodna istraživanja
pokazuju da procena klasičnih parametara fertiliteta ejakulata (koncentracija, ukupan
broj, pokretljivost i morfologija spermatozoida) nisu dovoljni pokazatelji fertiliteta i
reproduktivne performanse nerastova. Sa druge strane, pokazalo se da je fertilitet
veštački osemenjenih krmača, često, niži od onog kod prirodno osemenjenih krmača.
Kao osnovni razlog nižeg fertiliteta kod VO krmača, navodi se osemenjavanje sa
prekomerno razređenim dozama i/ili dozama dugotrajno čuvanim (3 do 5 dana).
Rezultati prethodnih istraživanja ukazuju da komponente semene plazme imaju ključni
uticaj na fertilizacioni potencijal spermatozoida in vivo i in vitro, kao i na fiziološke
procese važne za uspešnu oplodnju i razvoj embriona u uterusu.
Zbog toga je glavni cilj ovog istraživanja bio da se: (a) odredi sadržaj proteina u
spermalnoj plazma nerastova koji se koriste za VO na nekoliko komercijalnih farmi
svinja u Srbiji, (b) oceni uticaj sadržaja proteina u spermalnoj plazmi na pokretljivost i
morfologija spermatozoida u nativnoj i razređenoj spermi, nakon 3 dana čuvanja u
razređenom stanju i (c) ispita uticaj intrauterine infuzije spermalne plazme, pre
aplikacije klasične VO doze, na fertilitet krmača.
Sadržaj proteina u spermalnoj plazmi se kretao između 1% i 6,5%, što je
ustanovljeno kod 212 uzoraka, dobijenih iz ejakulata 106 nerastova, koji se koriste za
VO na 6 farmi u AP Vojvodini. Nizak nivo proteina (1-3.5%, prosečno 2,4%) je
ustanovljen kod 69%, a visok nivo proteina (3.6-6.5%, prosečno 4,2%) kod 31%
ispitivanih nerastova. Nije ustanovljen značajan (P>0,05) uticaj rase ili starosti nerasta,
kao ni godišnje sezone, na sadržaj proteina u spermalnoj plazmi. Citomorfološka
svojstva spermatozoida su testirana sistemom CASA i protočnom citometrijom. U
nativnoj spermi testiranih nerastova, prosečno je bilo 71% živih, 13% spermatozoida sa
oštećenim akrozomom i 32% spermatozoida sa morfološkim anomalijama. Volumen
ejakulata, koncentracija, ukupan broj i pokretljivost spermatozoida bili su značajno
(P<0,01) veći kod nerastova sa visokim, u poređenju sa niskim sadržajem proteina u
spermalnoj plazmi. Progresivna pokretljivost - PP (64%) i broj živih spermatozoida - ŽS
(66%) bio je značajno veći, dok su broj spermatozoida sa oštećenom ćelijskom
membranom - OM (19%), akrozomom - OA (29%) i hromozomima - OH (13%) bili
značajno (p<0,01) niži u uzorcima sperme sa visokim sadržajem proteina, koji su bili
čuvani 72h u razređenju 1:4, od ovih vrednosti kod uzoraka sa niskim sadržajem
proteina (PP = 48%, ŽS = 44%, OM = 27% OA = 45% i OH = 22%). Zamena autologne
spermalne plazme iz ejakulata sa niskim sadržajem proteina od jednog nerasta, sa
homologom spermalnom plazmom iz ejakulata drugog nerasta sa visokim sadržajem
proteina, značajno (p<0,01) povećava progresivnu pokretljivost spermatozoida, sa 52%
na 65%, kod uzoraka čuvanih 72h u razređenju 1:4. Intrauterina infuzija 30 ml semene
plazme, pre aplikacije klasične VO doze, značajno (p<0,05) povećava vrednost
prašenja (94%) i prosečan broj živo rođene prasadi po leglu (12.3) (p<0,01), u
poređenju sa kontrolom grupom krmača (83% i 10.5 prasadi).
Na osnovu dobijenih rezultata, može se zaključiti: (1) postoje značajne razlike u
sadržaju proteina u spermalnoj plazmi između pojedinih nerastova, (2) uzorci sperme sa
visokim sadržajem proteina, imaju veće vrednosti fertilizacionog potencijala
spermatozoida, od uzoraka sa niskim sadržajem proteina u spermalnoj plazmi, posle 72h
čuvanja u razređenju 1:4 i (3) infuzija spermalne plazme, pre aplikacije klasične VO
iv
doze, značajno povećava fertilitet tako tretiranih krmača, u poređenju sa kontrolnim
krmača. Ovi rezultati pokazuju da određivanje sadržaja proteina u spermalnoj plazmi,
može biti korisno sredstvo za predviđanje stepena fertiliteta nerasta, pre njegove
upotrebe za VO, kao i da se spermalna plazma može koristi za povećanje fertiliteta
veštački osemenjenih krmača. Dobijenim rezultatima su, u potpunosti, potvrđene radne
hipoteze i ostvareni postavljeni ciljevi istraživanja.

The ability to accurately evaluate the quality and estimate the fertilizing potential of dog semen has increased in importance as a result of the more widespread use of artificial insemination but remains challenging. The actual conception rate (CR) of a particular male of this polytocous species may provide useful information particularly in a competitive setting of a multi-sire mating or insemination. A crucial part in establishing the actual CR of a male is played by the fecundity of the female-which may ovulate from multi-ovular follicles-as well as breeding timing, breeding technique, and number of spermatozoa inseminated. This thesis determined, in a multi-sire insemination trial using DNA analysis and paternity testing, that the optimal day for surgical insemination using frozen-thawed semen was six days after the rise of the plasma progesterone concentration to between 6 and 9 nmol/L. Concurrently, the frozen-thawed semen used in the insemination trial was evaluated by means of conventional and modern semen evaluation methods, one of which, namely the Merocyanine 540 staining method, was newly validated on fresh dog sperm. Conventional assessment of sperm quality variables included individual progressive motility, viability and morphology using eosin-nigrosin staining. More modern semen evaluation procedures such as epifluorescence microscopy and flow cytometry were used to assess viability (Ethidium Homodimer, Yo-Pro 1), capacitation status (Anti-phosphotyrosine Clone 4G10), membrane destabilization (Merocyanine M540), acrosomal status (FITC-PNA), presence of progesterone receptors (P-BSA-FITC), motility parameters assessed using CASA, as well as the defragmentation index of sperm chromatin (SCSA). Males were ranked according to their CR which was then correlated to 40 sperm quality variables. Two sperm subpopulations, namely the percentage of live sperm which show signs of membrane destabilization (negatively), and the ability of sperm to maintain their viability (positively), did correlate or tended to correlate to in vivo fertility of the males. Another aspect of this thesis estimated the overall probability of a bitch having more than one conceptus derived from a smaller number of follicles, by retrospectively evaluating data of fertility trials as well as collecting data from private practice and welfare organization, thus establishing that the number of corpora lutea of a bitch may be used as a measurement for her fertility, despite the occurrence of multi-ovular follicles in the bitch. The current thesis assessed different aspects of male and female fertility in the domestic dog which, used in conjunction, may increase the ability to accurately estimate the fertilizing potential of frozen-thawed dog semen.
Thesis (PhD)--University of Pretoria, 2017.
Production Animal Studies
PhD
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La finalidad del presente estudio fue evaluar en 2 tiempos la calidad espermática de muestras de semen de ovino tratadas por la técnica de gradiente de densidad con respecto a muestras no tratadas. Se utilizó un total de 102 eyaculados de carnero que presentaron en promedio un volumen de 1.12 ± 0.34 ml, color blanquecino, aspecto cremoso, pH de 6.73 ± 0.25, concentración inicial de 3.54 x 109 ± 5.30 x 108 esp/ml y motilidad masal alrededor del grado 4. Posteriormente, cada eyaculado se dividió en un grupo control y experimental de volúmenes iguales. El semen del grupo control fue diluido con Triladyl® y el experimental, tratado con la técnica de gradiente de densidad. La tasa de recuperación espermática después del tratamiento fue del 31.50 ± 10.89%. Se encontró un aumento significativo (p<0.05) en la motilidad individual progresiva, el porcentaje de espermatozoides vivos y de espermatozoides con membrana intacta en las muestras tratadas con respecto al grupo control (92.07 ± 1.81% vs. 85.74 ± 1.75%, 78.42 ± 5.13% vs. 75.19 ± 4.59%, 78.55 ± 4.34% vs. 73.37 ± 4.48%, respectivamente); así como una disminución significativa (p<0.05) en el porcentaje de espermatozoides anormales en las muestras del grupo experimental con respecto al grupo control (8.41 ± 1.33% vs. 9.94 ± 1.73%). El resto de muestras de ambos grupos fue distribuido en pajillas de 0.25 ml y enfriadas hasta alcanzar los 5°C para su refrigeración por 24 horas. Las muestras tratadas con gradiente de densidad presentaron un aumento significativos (p<0.05), con respecto al grupo control, en motilidad individual progresiva, el porcentaje de espermatozoides vivos y el porcentaje de espermatozoides con membrana intacta (86.94 ± 3.14% vs. 82.44 ± 2.26%, 71.24 ± 4.11% vs. 68.82 ± 4.15%, 70.87 ± 3.11% vs. 67.98 ± 4.42%, respectivamente). En conclusión, la técnica de gradiente favoreció la obtención de un mayor número de espermatozoides vivos, de mejor motilidad y con membrana intacta, así como la disminución del número de espermatozoides anormales tanto en muestras frescas como refrigeradas. The purpose of the present study was to assess at 2 evaluation times the sperm quality from ram semen treated with the density gradient technique and to compare it against non-treated ram semen. A total of 102 ejaculates were evaluated. The mean macroscopic parameters were: volume of 1.12 ± 0.34 ml, creamy white color, creamy-dense appearance, pH of 6.73 ± 0.25, initial concentration of 3.54 x 109 ± 5.30 x 108 spz/ml and mass motility around grade 4. Then, each ejaculate was divided into two equal volume groups: the control and experimental group. For the control group, the sample was diluted in Triladyl®. The experimental group was treated with the density gradient technique. The recovery rate after treatment was 31.50 ± 10.89%.The results showed that, at zero hour, samples treated undergo a significant increase (p <0.05), compared with the control group, in individual progressive motility, percentage of living spermatozoa and the percentage of spermatozoa with intact membrane (92.07 ± 1.81% vs. 85.74 ± 1.75%, 78.42 ± 5.13% vs. 75.19 ± 4.59%, 78.55 ± 4.34% vs. 73.37 ± 4.48%, respectively). There was also a significant decrease (p <0.05) in the percentage of abnormal in samples from the experimental group compared to control group spermatozoa (8.41 ± 1.33% vs. 9.94 ± 1.73%). The remaining samples, from both the control and experimental group, were distributed in 0.25 ml straws and cooled to 5°C for 24 hours refrigeration. Then, the samples were evaluated. After 24 hours at 5°C, the samples treated with the density gradient technique showed significantly greater values (p <0.05), compared with the control group, in individual progressive motility, percentage of living spermatozoa and the percentage of spermatozoa with intact membrane (86.94 ± 3.14% vs. 82.44 ± 2.26%, 71.24 ± 4.11% vs. 68.82 ± 4.15%, 70.87 ± 3.11% vs. 67.98 ± 4.42%, respectively). In conclusion, in the present study, the density gradient technique allowed to obtain better a greater number of living spermatozoa with better motility, intact membrane and with lower abnormalities in fresh and refrigerated samples.

The objectives were to: 1) determine if im treatments of Lutalyse expedited the training of sexually inexperienced boars for semen collection and increased spermatozoal output, and 2) determine the effects of dietary L-carnitine supplementation on boar libido, semen quality, sperm production, and maintenance of sperm motility during liquid storage. Experiment 1 utilized lean-type, terminal-line boars (National Pig Development, Roanoke Rapids, NC) (n = 40; 177.4 ± 2.4 d of age and 112.8 ± 2.0 kg body weight) that had not previously experienced natural mating. Boars were individually moved twice weekly for 6 weeks (total of 12 training sessions) to a semen collection room equipped with an artificial sow. Upon entering the semen collection room, boars received in treatments of either deionized water (4 mL, n = 10) or Lutalyse at doses of 5 mg (n = 10), 10 mg (n = 10), or 20 mg (n = 10), and subsequently received a libido score of 1 to 5 (1 = no interest in the artificial sow; 5 = mounting the artificial sow and allowing semen collection). The percentages of boars successfully trained for semen collection during the experimental period were similar (P > 0.05) for controls (20%) and boars receiving 5 mg (30%), 10 mg (20%), or 20 mg (10%) of Lutalyse. Average libido score for boars receiving 10 mg Lutalyse (2.35 ± 0.08) was greater (P < 0.05) than for controls (2.14 ± 0.06). Libido score for the 20 mg treatment group were (1.78 ± 0.06) lower (P < 0.05) compared to the other treatment groups. Characteristics of ejaculates (volume, gel weight, sperm concentration, total spermatozoa) from control boars and boars treated with Lutalyse at doses of 5, 10, or 20 mg were similar (P > 0.05). For Exp. 2, the same group of boars was utilized in two similar trials (Trial 1, 1a, 1b: n = 9 for control and L-carnitine-treated boars; Trial 2, 2a, 2b: n = 10 for control and L-carnitine-treated boars). Boars were fed a fortified, corn and soybean meal-based diet at a rate of 2 kg/d. Boars that were randomly selected for L-carnitine treatment received the same diet mixed with L-carnitine to achieve supplementation of 500 mg/d. For 16 wk, semen was collected weekly via the gloved hand method and was analyzed for gel-free volume, gel weight, sperm concentration, sperm per ejaculate, and characteristics of sperm motility. Time to ejaculation (reaction time), duration of ejaculation, and number of false mounts were also recorded for each collection. Trials 1a and 2a were conducted during weeks 16 and 17 for each respective trial. Boars were collected once on 4 consecutive days, allowed 4 d of rest, and then collected again, to estimate daily spermatozoal production. At the end of 16 wk, a semen sample was also processed and extended in Beltsville Thawing Solution (BTS) to achieve a dilution of 3 x 109 spermatozoa/100 mL-dose for Trials 1b and 2b. The extended semen was stored in plastic bottles at 18°C and motility was evaluated daily for 7 d post collection. L-carnitine supplementation for 16 wk had no effects on semen volume, gel weight, total number of sperm cells per ejaculate, reaction time, or sperm motility (P > 0.1). Boars receiving the L-carnitine-supplemented diet displayed an increase in the number of false mounts before ejaculating and an increase in sperm concentration (P < 0.05) in Trial 2. A treatment by week interaction was detected for sperm concentration in Trial 2 (P < 0.005). Increased sperm concentrations in L-carnitine-treated boars were demonstrated after only one week of feeding the respective diets. Given that the production of a mature sperm cell requires 7 to 8 wk in boars, it is therefore difficult to conclude that differences in sperm concentration were due solely to treatment. Daily spermatozoal production was similar between control boars and boars supplemented with L-carnitine (P > 0.1) for both Trials 1a and 2a. L-carnitine supplementation did not affect percent motility in Trials 1b and 2b or sperm progressive motility in Trial 2b during 7 d storage (P > 0.1). A treatment by day interaction was determined for sperm velocity (P < 0.05) in Trial 2b. L-carnitine supplementation decreased mean sperm velocity significantly after 2 d of storage. Overall, L-carnitine had no beneficial effects on boar libido, semen quality, sperm production, or maintenance of sperm motility during liquid storage. However, Lutalyse increased libido scores, but did not affect the number of boars trained for semen collection or number of spermatozoa ejaculated.
Master of Science

[Truncated abstract]Judgements of physical attractiveness are thought to reflect evolved preferences for a high quality mate. The central aim of this thesis was to investigate the hypothesis that female preferences are adaptations for finding good quality mates and that faces and bodies signal honest information about mate quality. To date, most human mate preference studies have examined face or body attractiveness alone, and many have created stimuli using computer graphic techniques. Throughout these studies, I endeavoured to maximise the biological relevance of my studies by incorporating both face and body attractiveness, and using photographs of individual participants. Most research on attractiveness has focused on faces or bodies separately, while our preferences have evolved based on both seen together. A fundamental requirement of studying face and body attractiveness independently is that there is no interaction between the two. My first study confirmed that the face and body did not interact when an overall attractiveness judgment was made. I also investigated the independent contributions of rated attractiveness of the face and the body to ratings of overall attractiveness. Face and body attractiveness each made significant independent contributions to overall attractiveness in males and females. For both sexes, facial attractiveness predicted overall attractiveness more strongly than did body attractiveness, and this difference was significant in males. ... This study indicates that although current levels of T covary with male mating success, this effect may not be mediated by women's preferences for visual cues to T levels conveyed in static face or body features. The fourth study in this thesis investigated the phenotype-linked fertility hypothesis, which proposes that females obtain reliable information on male fertility from male expression of sexual traits. A previous study of Spanish men reported that facial attractiveness was positively associated with semen quality. I aimed to determine whether this effect was widespread by examining a large sample of Australian men. I also extended my study to determine whether cues to semen quality are provided by components of attractiveness: masculinity, averageness, and symmetry. I found no significant correlations between semen quality parameters and attractiveness or attractive traits. While male physical attractiveness may signal aspects of mate quality, my results suggest that phenotype-linked cues to male fertility may not be generalised across human populations. Together, these studies challenge current methodologies and theories of preferences for secondary sexual traits as honest signals of mate quality. The findings show that it is important to study human mate preferences in biologically relevant contexts, for example by using photographs of both faces and bodies, to maximise the real life application of results. In addition, the findings suggest that male attractiveness does not signal cues to testosterone or semen quality, although testosterone is associated with mating success. The implications of these findings and possible avenues for future research are discussed.

Equine arteritis virus (EAV) is the causal agent of equine viral arteritis (EVA), a disease of equids. Following EAV infection, up to 70% of stallions may become carriers and continuously shed the virus in their semen for varying time periods. The long-term carrier stallion has an important role in the transmission and maintenance of EAV in horse populations. Recently, it has been demonstrated a correlation between in vitro susceptibility of CD3+ T lymphocytes to EAV infection and establishment of long-term persistent infection among stallions following natural infections. In this study, we investigated whether stallions with in vitro EAV susceptible CD3+ T lymphocytes are at higher risk of becoming long-term carriers compared to those with the resistant phenotype following experimental infection with the KY84 strain of EAV. Furthermore, we investigated whether there is a significant effect of EAV infection on semen quality during acute phase of the infection. The data suggested that the establishment of the long-term carrier state seems to be associated with the in vitro CD3+ T lymphocyte susceptible phenotypes and that reduced semen quality resulted from the combined effect of fever and scrotal edema observed following EAV infection rather than the direct effect of the virus.

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Although more than 12 million doses of bull frozen semen were sold in Brazil in 2012, the production process of the doses sold by the artificial insemination centers registered in the Ministry of Agriculture, Livestock and Supply is not yet standardized. The hypothesis of the present study is that the design and implementation of a hazard analysis and critical control points (HACCP) system in a bull artificial insemination center can identify steps potentially harmful to the viability of semen doses, to decrease their rejection and to improvement their final quality, after the adoption of preventive and process control measures. The implementation of the HACCP system identified hazardous steps for the quality of the produced semen doses, leading to a reduction in their contamination during processing (P<0.05). There was also an increase in progressive motility and plasma membrane and acrosome integrity of spermatozoa (P<0.05) after implementing the HACCP system, which resulted in reduced rejection of semen batches and doses (P<0.05) and decreased production costs. Thus, the execution of the HACCP system standardized the production process of bull frozen semen, with improved sperm quality and reduction in both rejection of doses and costs to the industry. Additionally, during the sandwich doctorate program at the University of Calgary (Canada), the involvement of angiotensin converting enzyme in the capacitation process and in vitro fertilization in cattle was evaluated. After blocking the enzyme s activity by the specific inhibitor captopril, there were similar levels of tyrosine phosphorylation and cleavage and embryonic development until the blastocyst stage in comparison with the control group (P>0.05). Therefore, the present study suggests that the angiotensin converting enzyme is neither required for sperm capacitation nor involved in in vitro fertilization in cattle.
Apesar de o Brasil ter comercializado mais de 12 milhões de doses de sêmen bovino no ano de 2012, ainda não existe um padrão de processamento das doses comercializadas pelos estabelecimentos cadastrados pelo Ministério da Agricultura, Pecuária e Abastecimento. A hipótese do presente trabalho é de que a elaboração e a implantação de um sistema de Análise de Perigos e Pontos Críticos de Controle (APPCC) para um centro de coleta e processamento de sêmen bovino seja capaz de identificar as etapas potencialmente danosas para a viabilidade das doses, diminuir a sua rejeição e melhorar a sua qualidade final, a partir da adoção de medidas preventivas e de controle do processo. A implantação do sistema APPCC identificou as etapas que apresentavam perigos para a qualidade das doses de sêmen produzidas, ocorrendo redução na contaminação durante o seu processamento (P<0,05), Também houve um incremento na motilidade progressiva e na integridade das membranas plasmática e acrossomal dos espermatozóides (P<0,05) após a implantação do sistema APPCC, bem como uma redução na rejeição de partidas e doses de sêmen (P<0,05), o que se refletiu em uma diminuição dos custos de produção. Assim, a utilização do sistema APPCC resultou na padronização do processo de produção de sêmen bovino congelado, com incremento da qualidade seminal, redução na rejeição de doses e nos custos para a indústria. Adicionalmente, durante o programa de doutorado sanduíche na Universidade de Calgary (Canadá),foi avaliado o envolvimento da enzima conversora da angiotensina no processo de capacitação espermática e fertilização in vitro em bovinos. Após o bloqueio da atividade da enzima com o inibidor específico captopril,observou-se níveis semelhantes de fosforilação da proteína tirosina e de clivagem e desenvolvimento embrionário até o estágio de blastocisto, em comparação com o controle (P>0,05). Portanto, o presente estudo sugere que a enzima conversora da angiotensina não é necessária para o processo de capacitação espermática nem está envolvida na fertilização in vitro, em bovinos.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico
This dissertation is compounded for four trials. Experiment 1: This paper describes field results of some sexual behavior characteristics shown by donkeys when using an estrous mare for semen collection. Six donkeys (J: 1, 2, 3, 4, 5, 6) (Pêga breed), 3.5 16 year old and body weight from 230 to 330 kg. The jacks were conditioned to breed mares in a stud farm of mule production in Guacariaba, Minas Gerais, Brazil. For the semen collections, mares estrous signs were confirmed by a teaser stallion. The collections were done with an artificial vagina (Botucatu model), (water temperature 51° C) by the same veterinarian and far from the other donkeys. The collection intervals were from 48 to 72 h totaling 180 collections (17 to 40 semen collection/ breeder).The physical and morphological seminal parameters and were evaluated using a light microscope; gel fraction and semen volume were measured separated. Behavior data were analyzed by Kruskal Wallis test P < 0.05 by Statistical Analysis Systems-SAEG 9.1 (2007). The seminal parameters were: total motility 84.2 ± 6.0%, progressive motility 74.5 ± 7.1%, vigor 3.9 ± 0.5, semen volume 47.3 ± 28.7 ml and gel fraction 71.7 ± 54.8 ml. Gel fraction was present in 90/180 collections, in J1 39/40, but never in J4 and J5. The characteristics of sexual behavior were: reaction time 18.3 ± 17.6 min, time to the mount 5.1 ± 3.5 s, time to ejaculation 25.4 ± 7.7 s, number of mounts without erection1.1 ± 1.3 and number of Flehmen responses 7.4 ± 5.8. The older jacks group was the shorter was the reaction time and less mounts without erection. None of the sexual behavior characteristics was significantly different among jacks (P>0.05). All the collections started with sniffing of external genitalia. The ejaculation time, Flehmen response, and mounting without erection were similar to that described in natural breeding of jennies. However, no clapping mouth response was observed in the donkeys, neither difference in the mount time between jacks. In conclusion the used estrous mare was an efficient method of semen collection in donkeys neither significant effects on the sexual behavior. Experiment 2: The present study was performed with six jacks of the Pêga breed, weight from 231 to 326 kg (272 ± 34, 97). The breeders were divided in into the adult group (J1, J2 e J3, mean 15 years old) and a young group (J4, J5 e J6, mean 3,5 years old). The jacks presented records of normal rates of fertility at natural mating. Afterwards breeding soundness evaluation was performed and all animals ensure they were of capable for reproduction. Hundred eighty semen collections were performed, and the jacks presented normal seminal parameters for the specie. From August/2006 to February/2007, after semen collection each jack was submitted to three or four measurements of the extern genital tract (both right and left side) for: testis length (TL cranio caudal); testis height (TH dorsal ventral); testis width (TW lateral-lateral) and thickness of spermatic funiculum (FUN third part). The biometric data from testis biometric were used for calculation of testis indices (TI) and testis volume (TV). The average of TI observed was 7,45 (5,94 a 9,64) and TV 155,51 ± 14,45 149,34 ± 14,92 respectively left and right. The mean of FUN were for the right 25,35 ± 3,38 mm (20,38 a 30,80 mm) and 24,61 ± 1,59 mm (22 a 26 mm) for the left side. The jacks presented lesser TI than those proposed for stallions, but the asinine were more efficient than average related to stallion for sperm production. The testis volume associated with IT can be used in addition to the evaluation of the normality of the jacks breeder s testis. The measurement in vivo of spermatic funiculum can be used with other new component for the breeding soundness evaluation of jacks. The biometric values of the genital extern tract of this study it can support routine breeding evaluation and be applied to news studies. Experiment 3: This study was development in stud farm for mule and donkey production aiming to establish normal values of seminal parameters of jacks Pêga breed. Six jacks are used and arrangement in two groups adults (16, 15, 14 years old) an young (3.5, 3.5, 3.5 years old) weighting 272 ± 34,97 kg (231 326 kg). The animals were submitted to breeding soundness evaluation and all were approved for reproduction, beside that all animals present record of normal fertility. Were performed 180 semen collections (17 to 40/ jacks) by use artificial vagina Botucatu model. The values registered for physical seminal parameters and sperm morphology were: semen volume 47,27 ± 28,66 mL, gel fraction volume 71,75 ± 54,80 mL, total motility 84,22% ± 6,04, progressive motility 74,47 ± 7,06%, spermatic vigor 3,87 ± 0,51, sperm concentration 254,64 ± 91,25 x106/mL e, total sperm 10,3 ± 4525, 26 x 109, major spermatic defects and minor defects were 7,97 ± 3,02 % e 6,80 ± 1,59% in order. Were observed effect of class about seminal parameters, despite of the presence gel fraction was measurable in 30% of semen collections (54/180 collections) and the volume of gel fraction were influenced individually. This data may be used as references values for new studies. However, fields records and new studies involving greater number of breeders it should be done to establish real values for the specie. Experiment 4: This study comprehend the evaluation of the semen freezing methodology for jacks, with the application f the curve cooling pre freezing, and the use of two semen dilutors: Martim et al. (1979) and Nagase & Niwa (1964) modified. Twenty five semen collection were performed of five jacks Pêga breed; weighting 272 ± 34,97 kg and with age vary from 3,5 years to 17 years. Evaluations were performed in raw semen, cooled, and freezing semen in different phases of semen cryopreservation of the jacks Pêga breed. The analyses were: sperm morphology, membrane integrity by test stress hyposmotic and supravital stain and also physical seminal parameters. In addition, was realized fertility test in 51 mares (60 estrous cycles), 30 estrous cycles/dilutor with the semen from three breeders. The pregnancy rates were 13, 25 and 35 days were: 53,33%, 43,66% e 16,6% for Nagase & Niwa (1964) modified and 50%, 46,66% e 10% Martim et al. (1979).The hiposmotic test showed suitable test for indication the sperm cells survivors to the cryopreservation procedures, while the supravital stain was unsuitable for the present methodology. The pregnancy rates were elevated between days 13th and 25th, but slight rates of embryo loss. The traditionally dilutors used for semen bulls and stallions can be used for jacks semen cryopreservation.
Esta dissertação compreende quatro experimentos. Experimento1: Este estudo descreve algumas características do comportamento sexual de jumentos quando uma égua em estro foi usada para a coleta de sêmen. Foram usados neste estudo seis jumentos (J: 1, 2, 3, 4, 5, 6) da raça Pêga, com idade variando de 3,5 a 16 anos e pesando de 230 a 330 kg. Os animais foram previamente condicionados em um haras comercial de produção de muares localizado em Guaraciaba, Minas Gerais, Brasil. Para a colheita de sêmen, as éguas tiveram seu estro confirmado com o uso de um garanhão. As coletas foram realizadas com vagina artificial (modelo Botucatu), com água aquecida a 51ºC, pelo mesmo médico veterinário e preferencialmente distante de outros asininos. As coletas foram realizadas em intervalos de 48 a 72 horas, totalizando 180 coletas (17 a 40 coletas/reprodutor). Os parâmetros físicos e morfológicos do sêmen foram avaliados empregando-se um microscópio óptico comum, e a fração gel e a fração livre de gel foram mensuradas e avaliadas separadamente. Os dados de comportamento sexual foram analisados com uso do teste de Kruskal Wallis (P < 0.05), com o Programa de Análises Estatísticas e Genéticas SAEG 9,1 (UFV, 2007). Os parâmetros seminais foram: motilidade total 84.2 ± 6.0%, motilidade progressiva 74.5 ± 7.1%, vigor 3.9 ± 0.5, volume sêmen 47.3 ± 28.7 mL e a fração gel de 71.7 ± 54.8 mL. A fração gelatinosa esteve presente em 90 das 180 coletas, e para J1 39/40 (relação total de coletas), porém J4 e J5 não apresentaram essa fração. As características do comportamento sexual foram: tempo de reação 18.3 ± 17.6 minutos; tempo de monta 5.1 ± 3.5 segundos; time de ejaculação 25.4 ± 7.7 segundos; número de montas sem ereção 1.1 ± 1.3 e número de resposta de Flehmen 7.4 ± 5.8. Os jumentos adultos (J1, J2, J3) apresentaram menor tempo de reação em relação aos demais e menor número de montas sem ereção. Não foram observadas diferenças significativas entre jumentos no seu comportamento (P>0.05),porém foi observada diferenças entre grupos quanto ao tempo de reação. Todas as coletas de sêmen foram iniciadas com investigação olfativa do genital externo das éguas. O tempo de ejaculação, resposta de Flehmen e montas sem ereção foram similares ao descrito em monta natural usando jumentas. Entretanto, não foi observado comportamento mastigatório exibido por alguns reprodutores na presença de jumentas. O uso de égua em estro para coleta de sêmen pode ser feito eficientemente sem afetar significativamente o comportamento sexual. Experimento 2: Este experimento foi conduzido com seis jumentos reprodutores da raça Pêga, com peso médio de 272 ± 34,97 (231 326 kg), sendo subdivididos em adultos (J1, J2 e J3, 16,14 e 15 respectivamente) e jovens (J4, J5 e J6, média 3,5 anos). Os reprodutores apresentavam histórico de fertilidade normal à monta natural, e após realização de exame andrológico foram classificados como aptos à reprodução. Foram realizadas 180 coletas de sêmen, tendo os reprodutores apresentado parâmetros seminais considerados normais para a espécie. De agosto/2006 a fevereiro/2007, cada reprodutor, após a coleta de sêmen, foi submetido a três ou quatro mensurações do aparelho genital externo para comprimento testicular (CT - crânio caudal), altura testicular (AT dorso ventral), largura testicular (LT médio lateral) e espessura do funículo (FUN terço médio). Os dados de biometriatesticular foram utilizados para o cálculo do índice testicular (IT) e volume testicular (VT). O IT médio observado foi de 7,45 (5,94 a 9,64) e o VT esquerdo foi de 155,51 ± 14,45 mL, enquanto o direito foi de 149,34 ± 14,92. A medida de FUND (funículo espermático direito) foi de 25,35 ± 3,38 mm (20,38 a 30,80 mm) e a de FUNE (funículo espermático esquerdo) foi 24,61 ± 1,59 mm (22 a 26 mm). Jumentos, mesmo com IT menor do que o proposto para garanhões; se apresentaram mais eficientes que os últimos, para a produção espermática. O volume espermático associado ao IT pode ser usado como objeto auxiliar na estimativa da constatação de normalidade para testículos de jumentos. A mensuração in vivo do funículo espermático pode ser uma nova variante na utilização do exame andrológico de jumentos. Os dados descritos podem servir de valores de referência e valores práticos quanto à biometria do aparelho genital externo de jumentos. Experimento 3: Este experimento foi conduzido em um haras comercial de produção de muares. Utilizaram-se seis jumentos reprodutores agrupados em adultos (16, 15, 14 anos) e jovens (3.5, 3.5, 3.5 anos), pesando 272 ± 34,97 (231 326 kg), com objetivo de determinar os parâmetros seminais para a raça Pêga. Os animais foram submetidos a exame andrológico, sendo classificados como aptos à reprodução. Realizou-se 180 coletas de sêmen (17 a 40/animal) com o emprego de vagina artificial (modelo Botucatu). Os resultados para os parâmetros físicos e morfológicos do sêmen foram: volume sêmen: 47,27 ± 28,66 mL; volume da fração gel: 71,75 ± 54,80 mL; motilidade total 84,22% ± 6,04; motilidade progressiva: 74,47 ± 7,06%; vigor espermático: 3,87 ± 0,51; concentração espermática: 254,64 ± 91,25 x106/mL; espermatozóides totais: 10,3 ± 4525, 26 x 109; defeitos espermáticos maiores e menores: 7,97 ± 3,02 % e 6,80 ± 1,59%, respectivamente. Foram observados efeitos de classe sobre os parâmetros seminais, sendo que a presença da fração gelatinosa foi detectável em 30% das coletas (54/180 coletas) e o volume gel foi influenciado individualmente. Estes dados podem servir de referências para novos estudos. Contudo, levantamentos de campo e novos estudos devem ser conduzidos envolvendo maior número de animais para que sejam construídos valores de referência para a espécie. Experimento 4: Este delineamento compreende a avaliação de uma metodologia de congelamento do sêmen de jumentos, com o emprego de uma curva de resfriamento prévio, e com uso de dois diluidores de congelamento: meio diluidor de Martim et al. (1979) e meio diluidor de Nagase & Niwa, (1964) modificado. Foram realizadas vinte e cinco coletas de sêmen com uso de vagina artificial de cinco reprodutores da raça Pêga, pesando em média 272 ± 34,97 e com idades variando de 3,5 -17 anos. Foram realizados avaliações do sêmen fresco, resfriado e congelado nas diferentes fases da criopreservação do sêmen analisando a morfologia espermática, integridade de membrana por coloração de eosina nigrosina e teste hiposmótico, além da avaliação subjetiva dos parâmetros físicos. Em adição, foi realizado teste de fertilidade com sêmen de três reprodutores em 51 éguas (60 ciclos), sendo 30ciclos/diluidor, com o objetivo de estudar alguns aspectos envolvidos na criopreservação do sêmen de jumentos da raça Pêga. As taxas de gestação aos 13, 25 e 35 dias foram: 53,33%, 43,66% e 16,6% Nagase & Niwa (1964) modificado e 50%, 46,66% e 10% Martim et al. (1979). O teste hiposmótico se mostrou adequado para indicar a população de células que sobrevivem ao congelamento, enquanto a coloração de eosina nigrosina se mostrou inapropriada na presente metodologia. As taxas de gestação foram altas aos 13 e 25 dias pós- inseminação, porém com elevadas taxas de perdas embrionárias. Os diluidores tradicionalmente empregados para sêmen de touros e garanhões podem ser empregados para a criopreservação do sêmen de jumentos.

[ES] Cualquier mejora en el conocimiento de la nutrición y el manejo de los futuros conejos reproductores debe considerarse como prioritario en la producción cunícola. Sin embargo, los principales esfuerzos en investigación se han centrado en estos animales durante la fase de reproducción propiamente dicha. La hipótesis inicial de esta tesis fue que alcanzar un desarrollo temprano adecuado durante la gestación, lactancia, engorde y recría, con la ayuda de programas ajustados de alimentación en algunos de estos períodos, mejoraría el rendimiento reproductivo y la vida útil de los futuros conejos reproductores. Para evaluar esta hipótesis, se llevaron a cabo dos tipos de ensayos, tanto en conejos hembra de líneas maternas como en machos de líneas seleccionadas por velocidad de crecimiento y destinadas a la inseminación artificial. El primero de estos ensayos se centró en los diferentes patrones de crecimiento, desde el nacimiento hasta el final de la fase de recría, y su influencia en la carrera reproductiva y la vida útil del conejo. En el segundo tipo de pruebas, se evaluó el efecto del uso de diferentes dietas o programas de alimentación durante el período de recría en la productividad a corto y medio plazo. Los resultados muestran dos fases importantes en el desarrollo temprano, de acuerdo con sus efectos sobre el rendimiento reproductivo y la vida útil. Mostrar un mayor desarrollo corporal desde el nacimiento hasta los 63 días de vida parece tener una influencia positiva en el grado de madurez y la productividad al comienzo de su vida reproductiva, además de poder mejorar su supervivencia a corto plazo, pero con posibles efectos negativos en la esperanza de vida a largo plazo. Sin embargo, el desarrollo durante el período de recría (desde los 63 días de vida hasta el comienzo de la carrera reproductiva) parece tener un mayor impacto en la productividad y la esperanza de vida de nuestros futuros conejos reproductores. Las hembras más engrasadas en la primera inseminación tuvieron tamaños de camada más pequeños y un mayor riesgo de ser sacrificadas que las magras (P<0.05). Además, los machos que mostraron una mayor ganancia media diaria durante el período de recría tuvieron un número y porcentaje de eyaculados viables más bajos a lo largo de su vida productiva y mayor riesgo de muerte o sacrificio. Estos resultados podrían indicar que la clave para llevar a cabo un desarrollo adecuado durante el período de recría debería basarse tanto en conseguir que los conejos alcancen un grado de madurez apropiado, tanto fisiológica como reproductivamente, así como evitar un excesivo engrasamiento al inicio de su vida productiva. En los ensayos destinados a evaluar los programas de alimentación durante la recría, el uso de una restricción alimentaria en hembras, y especialmente el uso de dietas fibrosas (de 8.5 a 10 MJ ED/kg MS) permiten un desarrollo corporal gradual y adecuado a la edad de cubrición, con suficientes reservas corporales para afrontar el comienzo de la vida reproductiva y mejorando tanto su rendimiento reproductivo como su vida útil. El éxito de estos programas de alimentación de recría depende del momento y la duración de la aplicación de estos, mejorándose con la aplicación de un cambio a un pienso enriquecido energéticamente alrededor del primer apareamiento. En los machos, se observó que un programa de alimentación durante la recría adecuado debería adaptarse a la capacidad de ingesta de estos y a sus necesidades nutricionales en períodos concretos de la recría, consiguiendo así mejoras en algunos parámetros del semen al inicio de su vida reproductiva. En conclusión, tanto un desarrollo temprano adecuado desde la gestación hasta el comienzo de su vida reproductiva, como el uso de programas adecuados de alimentación durante la recría podrían mejorar significativamente el rendimiento reproductivo futuro, así como la esperanza de vida, tanto en conejos he
[CAT] Qualsevol millora en el coneixement tant de la nutrició com del maneig dels futurs conills de criança ha de considerar-se com a prioritari en la producció de conills. No obstant açò, els principals esforços en investigació s'han centrat en aquests animals durant la fase de reproducció pròpiament dita. La hipòtesi inicial d'aquesta tesi va ser que aconseguir un desenvolupament primerenc adequat durant la gestació, lactància, engreix i criança, amb l'ajuda de programes ajustats d'alimentació en alguns d'aquests períodes, milloraria el rendiment reproductiu i la vida útil dels futurs conills reproductors. Per a avaluar aquesta hipòtesi, es van dur a terme dos tipus d'assajos, tant en conills femella de línies maternes, com en mascles de línies seleccionades per a la velocitat de creixement i destinades a la inseminació artificial. El primer d'aquests assajos es va centrar en els diferents patrons de creixement, des del naixement fins al final de la fase de criança, i la seua influència en la carrera reproductiva i la vida útil del conill. En el segon tipus de proves, es va avaluar l'efecte de l'ús de diferents dietes o programes d'alimentació durant el període de criança, en la productivitat a curt i mig termini. Els resultats obtinguts semblen indicar que podríem diferenciar dues fases en el desenvolupament primerenc, d'acord amb els seus efectes sobre el rendiment reproductiu i la vida útil. Mostrar un major desenvolupament corporal des del naixement fins als 63 dies de vida sembla tenir una influència positiva en el grau de maduresa i la productivitat al començament de la seua vida reproductiva, a més de poder millorar la seua supervivència a curt termini, però amb possibles efectes negatius en l'esperança de vida a llarg termini. No obstant açò, el desenvolupament durant el període de sembla tenir un major impacte en la productivitat i l'esperança de vida dels nostres futurs conills reproductors. Les femelles més greixades en la primera inseminació van tenir grandàries de ventrada més xicotets (tant nascuts com deslletats) i un major risc de ser sacrificades que les magres (P<0.05). A més, els mascles que van mostrar un major guany mitjà diari durant el període de criança van tenir un nombre i percentatge d'ejaculats viables més baixos al llarg de la seua vida productiva i major risc de mort o sacrifici. Aquests resultats podrien indicar que la clau per a dur a terme un desenvolupament adequat durant el període de criança hauria de basar-se tant a aconseguir que els conills aconseguisquen un grau de maduresa apropiat, per a afrontar els futurs desafiaments productius, com evitar un excessiu greixatge a l'inici de la seua vida productiva. En els assajos destinats a avaluar els programes d'alimentació durant la criança, l'ús d'una restricció alimentària en femelles, i especialment l'ús de dietes fibroses (de 8.5 a 10 MJ ED/kg MS) permeten un desenvolupament corporal gradual i adequat a l'edat de cobriment, amb suficients reserves corporals per a afrontar el començament de la vida reproductiva i millorant tant el seu rendiment reproductiu com la seua vida útil. L'èxit d'aquests programes d'alimentació de criança depèn del moment i la durada de l'aplicació d'aquests, millorant-se amb l'aplicació d'un canvi a un pinso enriquit energèticament al voltant del primer cobriment. En els mascles, es va observar que un programa d'alimentació durant la criança adequat hauria d'adaptar-se a la capacitat d'ingesta d'aquests i a les seues necessitats nutricionals en períodes concrets de la criança, aconseguint així millores en alguns paràmetres del semen a l'inici de la seua vida reproductiva. En conclusió, tant un desenvolupament primerenc adequat des de la gestació fins al començament de la seua vida reproductiva, com l'ús de programes adequats d'alimentació durant la criança podrien millorar significativament el rendiment reproductiu futur, així com l'esperança
[EN] Any improvement on the knowledge of both nutrition and management of the future rabbit breeders should be considered as crucial. However, main research efforts have been focused on animals during reproduction phase. The initial hypothesis of this thesis was that achieving an adequate early development during young rabbits' gestation, lactation, fattening and rearing, with the help of fitted feeding programmes in some of these periods, would be improved reproductive performance and lifespan of the future breeding rabbits. To evaluate this hypothesis, two types of trials, both in rabbit females from maternal lines selected for litter size at weaning and males from paternal lines selected for growth rate and intended on artificial insemination, were carried out. The first of these trials were mainly focused on the different growth patterns, from birth to the end of the rearing age, and their influence on rabbit's reproductive career and lifespan. In a second type of trials, the effect of the use of different diets or feeding programs during the rearing period in the performance in the short- medium-term was evaluated. The results obtained from the evaluation of the different growth patterns trials seem to indicate that, we could differentiate two parts on the early development according to their effects on reproductive performance and lifespan. To show a greater body development from birth to 63 days of life seems to have a positive influence on the maturity degree and productivity at the beginning of their reproductive life, as well as being able to improve their survival in the short-term, but it could be even negative for long-term lifespan. However, the development during the rearing period (from 63 days of life to the beginning of reproduction career) seems to have a higher impact on the productivity and lifespan of our rabbit breeders. Fatter rabbit females at first insemination had smaller litter sizes (born and weaned) and a higher risk of being culled (+13% per positive change in mm of perirenal fat) than lean ones (P<0.05). Also, rabbit males that showed a greater average daily gain during rearing period had a lower number and percentage of profitable ejaculates through it productive life (-10.1 and -4.9% per each 10 g of daily gain increase, respectively; P<0.05) and higher risk of death or culling (+16.2% per each increase in one standard deviation unit on live weight; P<0.05). These results could indicate that the key to carrying out an adequate development during the rearing period should allow the rabbits to reach an appropriate degree of physiological and reproductive maturity to face the future productive challenges, avoiding an excessive fatness at the beginning of their productive life. The use of a feeding restriction in rabbit females, and especially the use of fibrous diets (from 8.5 to 10 MJ DE/ kg DM) lead a gradual and adequate body development at mating, with enough body reserves to face the beginning of reproduction life and improving their whole reproductive performance and lifespan. The success of these rearing feeding programs depends in the moment and duration of application of them and improved with the application of a nutritional flushing around first mating. In rabbit males, it was observed that an adequate rearing feeding program should adapt it to the male's intake capacity and their nutritional requirements rearing period, especially to their high DP needs at the beginning of rearing period or when their requirements were close to maintenance, to improve some semen parameters at the beginning of their reproduction life. In conclusion, both the adequate early development of future rabbit breeders, from their own gestation to the beginning of their reproductive life, and the use of appropriate rearing feeding programs could significantly improve their future reproductive performance, as well as their life expectancy, both in rabbit females and males.
Martínez Paredes, EM. (2019). Rearing programmes for breeding rabbits. Effect of early development and feeding on future reproductive performance of rabbit females and males [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/124969
TESIS

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The objective of this study was to study the influence of magnetized water intake on body weight gain, body morphometry and reproductive system, semen characteristics and body and scrotal area temperatures with digital infrared thermography in young Nellore bulls in extensive management and In confinement. Twenty bulls with initial age of 14 months were used, divided into two groups: Treated - water ingestion treated by magnetic field; And Control - drinking water intake. Four collections were made, 2 in extensive management (14 and 18 months of age) and 2 in confinement (20 and 21 months of age), in which they were measured: climatic factors, rectal temperature, body weight, wither height, Length, width and height of the testicles, semen collection and infrared digital thermography of the eyeball, mufla, flank, ischium and scrotum. It was concluded that the intake of water treated by magnetic field was not efficient in relation to weight gain in pasture and in confinement, as well as for body morphometry and reproductive system. In confinement, there was also no influence of water treated by magnetic field on the quantitative and qualitative characteristics of the semen. Digital infrared thermography proved to be effective for measuring body and scrotal areas, where the grazing group treated with magnetic field water showed higher temperatures for the ocular globe, mufla, flank and ischium. For the scrotal pouch, the pasture, the temperature of the spermatic cord and testis were higher in the treated group; And in confinement, the temperature of the testes and tail of the epididymis were higher in the group with drinking water intake.
Objetivou-se estudar a influência da ingestão da água magnetizada no ganho de peso corpóreo, morfometria corpórea e do aparelho reprodutor, características do sêmen e temperaturas de áreas do corpo e escroto com termografia digital de infravermelho em touros jovens da raça Nelore em manejo extensivo e em confinamento. Foram utilizados 20 touros com idade inicial de 14 meses, divididos em dois grupos: Tratado - ingestão de água tratada por campo magnético; e Controle - ingestão de água potável. Foram realizadas 4 coletas sendo 2 em manejo extensivo (14 e 18 meses de idade) e 2 em confinamento (20 e 21 meses de idade), nas quais foram aferidos: fatores climáticos, temperatura retal, peso corpóreo, altura de cernelha, mensuração do comprimento, largura e altura dos testículos, colheita de sêmen e termografia digital por infravermelho do globo ocular, mufla, flanco, ísquio e escroto. Conclui-se que a ingestão da água tratada por campo magnético não se apresentou eficiente em relação ao ganho de peso a pasto e em confinamento, assim como para morfometria corpórea e do aparelho reprodutor. Em confinamento, também não houve influência da água tratada por campo magnético sobre as características quantitativas e qualitativas do sêmen. A termografia digital por infravermelho se mostrou eficaz para mensuração das áreas do corpo e da bolsa escrotal, onde a pasto o grupo com ingestão de água tratada por campo magnético apresentou temperaturas superiores para globo ocular, mufla, flanco e ísquios. Para bolsa escrotal, a pasto, a temperatura do cordão espermático e testículo foram superiores no grupo tratado; e em confinamento, a temperatura do testículo e cauda dos epidídimos foram superiores no grupo com ingestão de água potável.

Após a inseminação artificial, as fêmeas apresentam reação inflamatória transitória e fisiológica, que pode resultar em alterações na vascularização uterina. A ultrassonografia Doppler colorido é uma técnica não invasiva, que vem sendo utilizada para estudar a hemodinâmica do trato reprodutivo de bovinos. Os objetivos deste trabalho foram verificar a resposta inflamatória uterina por ultrassonografia Doppler colorido e a taxa de fertilidade em bovinos após inseminação artificial (IA) com sêmen avaliado por sondas fluorescentes. Para isso foram delineados dois experimentos. O experimento 1 foi realizado para estudar a hemodinâmica uterina por ultrassonografia Doppler colorido em bovinos após a IA. Foram utilizadas 18 vacas da raça Nelore, submetidas a um protocolo para a inseminação artificial em tempo fixo (IATF) e divididas em dois grupos: Grupo não inseminado (controle, GC, n=9) e grupo submetido à inseminação artificial (GIA, n=9). Avaliações por ultrassonografia foram realizadas em cinco momentos em modo espectral (30 horas antes da IA, 4, 24, 48 e 168 horas após a IA) para avaliar o índice de resistência (RI) da artéria uterina e em quatro momentos em modo color Doppler (4, 24, 48 e 168 horas após a IA) para avaliar a vascularização dos cornos uterinos. O diagnóstico de gestação foi feito após 30 dias da IA. No experimento 2 objetivou-se verificar se o percentual de espermatozóides com integridade das membranas plasmática e acrossomal e função mitocondrial (PIAIC) avaliadas por sondas fluorescentes influencia a fertilidade e estudar a hemodinâmica uterina após a deposição do sêmen com diferentes percentuais de espermatozóides PIAIC. Foram utilizadas 182 vacas paridas da raça Nelore, submetidas a um protocolo para IATF e divididas em três grupos de acordo com a qualidade do sêmen: Bom (B) 44,5%, Médio (M) 23% e Regular (R) 8,5% de espermatozóides PIAIC/palheta. Uma amostra de 30% das fêmeas (54, sendo 18 de cada grupo) foram submetidas a avaliações com ultrassonografia Doppler colorido 30 horas antes da IA, 4 e 24 horas após a IA. Foram considerados os RI das artérias uterinas direita (AD) e esquerda (AE) e vascularização subjetiva dos cornos uterinos direito (CD) e esquerdo (CE). Posteriormente, foram separados levando em consideração o lado do ovário contendo o folículo dominante para comparar RI das artérias ipsi (AI) e contralateral (AC) e vascularização subjetiva dos cornos uterinos ipsi (CI) e contralateral (CC). O diagnóstico de gestação foi realizado 30 dias após a IA. Para a análise estatística foi utilizado o programa SAS versão 9.2 (SAS 2010). As variáveis foram analisadas pelo PROC MIXED, utilizando modelo linear misto para medidas repetidas no tempo. Foram considerados os efeitos fixos de grupo, tempo e interação grupo x tempo. Os dados obtidos foram submetidos à análise de variância e ao teste de Duncan, pelo PROC GLM. A análise de Qui-Quadrado foi utilizada para determinar a taxa de prenhez, de acordo com a qualidade do sêmen pelo PROC FREQ. O nível de significância considerado foi de 5%. No experimento 1 não foram observadas diferenças dos parâmetros avaliados por ultrassom Doppler entre GIA e GC, mas foram notados efeitos do tempo para RI, sendo mais baixos 4 horas após a IA. Quando os dados foram avaliados de acordo com o lado do folículo dominante foi observado que 4 e 24 horas após a IA, RI da AC do GIA foi mais baixo do que o GC. No experimento 2 não houve diferença entre os animais inseminados com sêmen B, M ou R quanto a hemodinâmica uterina, mas houve efeito do tempo, sendo que 4 horas após a IA verificou-se menor valor de RI e maior fluxo sanguíneo. A taxa de prenhez dos animais inseminados com o sêmen B (64,7%) foi maior quando comparada ao sêmen R (36,2%), sendo que ambos não diferiram do sêmen M (50,0%). Pode-se concluir que não é possível detectar por ultrassonografia Doppler alterações na hemodinâmica uterina devido à IA em fêmeas bovinas. A deposição de sêmen com diferentes qualidades não altera a hemodinâmica uterina de forma detectável por ultrassonografia Doppler em bovinos. Por outro lado, é possível detectar por ultrassonografia Doppler mudanças na perfusão sanguínea do útero em relação ao tempo da IA, o que pode ser devido a alterações hormonais que ocorrem durante o ciclo estral. Por fim, a IA com sêmen contendo menor percentual de espermatozoides PIAIC causa redução na taxa de fertilidade em bovinos.
After insemination, females have physiological and transient inflammatory reaction, which can result in changes in the uterine vasculature. The color Doppler ultrasound is a noninvasive technique that has been used to study the hemodynamics of cattle reproductive tract.The purposes of this work were to verify uterine inflammatory response by color Doppler ultrasound and fertility rate in cattle after artificial insemination (IA) with semen evaluated by fluorescent probes. For this, two experiments were delineated. The first experiment was conducted to study the uterine hemodynamic by color Doppler ultrasound in cattle after AI. Were used 18 Nellore cows that were subjected to a protocol for fixed timed artificial insemination (IATF). The animals were divided in two groups: Not inseminated group (control, GC, n=9) and inseminated group (GIA, n=9). Ultrasonografic evaluation were done in five moments on spectral mode (30 hours before AI, 4, 24, 48 e 168 hours after IA) to evaluate the resistence index (RI) of uterine artery and in four moments in color Doppler mode to evaluate the uterine horns vascularization. The pregnancy diagnostic was done 30 days after IA. In the second experiment the objective was to evaluate whether the percentage of sperm with plasma membrane integrity and acrossomal and mitochondrial function (PIAIC) assessed by fluorescent probes influences fertility and study the uterine hemodynamic after the deposition of semen with different percentages of PIAIC sperm. Were used 182 caved Nellore cows submitted to a protocol for IATF and divided into three groups according to semen quality: Good (B) 44.5%, Medium (M) 23% and Regular (R) 8,5% of PIAIC sperm/blade. A sample of 30% of female (54, 18 in each group) were submitted to evaluation by color Doppler ultrasound 30 hours before IA, 4 and 24 hours after IA. Were considered RI of right (AD) and left (AE) uterine arteries and subjective vascularization of right (CD) and left (CE) uterine horns. Later, they were separated taking into account the side of the ovary containing the dominant follicle to compare RI of ipsi (AI) and contralateral (AC) arteries and subjective vascularization of ipsi (CI) and contralateral (CC) uterine horns. The pregnancy diagnostic was done 30 days after IA. Statistical analysis was performed using the program SAS version 9.2 (SAS 2010). The variables were analyzed by PROC MIXED, using mixed linear model for repeated measures in time. We considered effects of group, time and group x time interaction. The data obtained were submitted to ANOVA and Duncan test, using PROC GLM. The chi-square analysis was used to determine pregnancy rate, according to the quality of semen by PROC FREQ. the significance level was 5%. In experiment 1 there were no differences of the parameters evaluated by Doppler ultrasound between GIA and GC, but time effects were noted for RI, it was lower four hours after IA. When the data were evaluated according to the side of the dominant follicle was observed that 4 and 24 hours after IA, RI of AC of GIA was lower when it was compared with GC. In experiment 2 there was no difference between animals inseminated with semen B, M or R about uterine hemodynamics, but there were effects of time, so that 4 hours after IA showed lower value of RI and increased blood flow. The pregnancy rate of animals inseminated with semen B (64.7%) was higher when compared with semen R (36.2%), both of them did not differ of semen M (50.0%). It can be concluded that is not possible to detect uterine hemodynamic changes by Doppler ultrasound due to IA in cattle. The deposition of semen with different qualities does not change the uterine hemodynamics enough to be detectable by Doppler ultrasonography in cattle. On the other hand, is possible to detect changes in blood perfusion of the uterus in relation to the time of IA by Doppler ultrasound, which may be due to hormonal changes that occur during the estrous cycle. Finally, IA with semen containing a lower percentage of PIAIC sperm causes a reduction in fertility rate in cattle.

Content: We are a group company of a major department store in Japan. We are in charge of quality control and customer service of items sold by our stores. We constantly monitor our customers’ concerns, and we have been keeping customer correspondence records since 1996. We hold over 100,000 cases for all items of food, clothing, living and services. The record includes not only customer requests but also test results of each item before sales as well as reproduction test results based on customer requests. We use the database to build our quality standards for everything we sell and for the education of our sales staff. All personal information in these correspondence records are, of course, kept completely confidential. However, improving the quality of the items we sell is not simply a problem of finding solutions within our supply chain. Using compiled results, we aim to clarify the type and level of problems users experience with leather goods, leading to overall quality improvement of supply items of scale. However, we cannot perform this task alone. As a survey method, it is based on compiling all 7,000 records of leather goods over the past decade sorted by our original ‘product-code’ and ‘consultation-code’. By using compiled results, we are able to clarify what kinds of items and what kind of complaints is more often in major sales items. Additionally, by using text extraction from the content of customers’ requests and compiling them manually, totals can be determined for each use period and product color. For example, we can compare customer complaint data with test results for colour fastness. Referring to required performance items specified by ISO standards, the gap between customer requests and quality standards can be clarified. With the internationalization of purchasing networks of sales items, quality control based on setting various safety and quality standards is becoming increasingly important. In terms of the characteristics of leather, we think that there is probably a level that is impossible to realize, however, not only the problems of regulations and safety but the nature of the customer's viewpoint must also be discussed. Take-Away: We are going to present current data showing proof of what consumers actually want. Using this data, leather goods manufacturers can efficiently make products that consumers want and avoid making products that consumers don’t want. We suggest that quality standards , including ISO, should consider not only safety and regulations of environment, but consumer`s opinions as well.

Insomnia is common in children, and is associated with decreased school performance and increased psychopathology. Although adult insomnia is linked to worsened health-related quality of life (HRQOL), there is insufficient data evaluating insomnia and HRQOL in children. We examined the HRQOL and health associations of insomnia in a longitudinal cohort of 194 children (96 girls, age at study start 8.7 +/- 1.6 years, age at data analysis 15.0 +/- 1.8 years) over 7 years. International Classification of Sleep Disorders, second edition (ICSD2) derived insomnia was seen intermittently in 27% of children, and was persistent in 4%. Children reporting ICSD2-derived insomnia had lower HRQOL. Additionally, the presence of insomnia was associated with an increased risk of reporting a new medical condition (intermittent insomnia odds ratio 5.9 [95% CI 1.3-26.7, p = 0.04], persistent insomnia odds ratio 8 [95% CI 2.3-27.7, p = 0.001]). Persistent ICSD2-derived insomnia was associated with an increased risk of reporting a new medication (odds ratio 4.9 (95% CI 1.0-23.6), p = 0.049), and reporting a new psychiatric medication (odds ratio 13.7, 95% CI: 2.6-73.5, p = 0.002). These associations were present even after adjusting for socioeconomic factors and the presence of obstructive sleep apnea. Insomnia in children is associated with worsened HRQOL and health outcomes.

Master’s thesis occupies with problems of quality in company Karsit L.L.C. First part is directed at analysis of present situation and at description of problems in a theory level. In next part tools of quality are applied at particular product and analysis of spoilage and process capability is executed.

Les aérosols sont une composante variable de l'atmosphère, ayant un rôle important et complexe sur la qualité de l'air et le climat. La plupart des observations d'aérosols au sol sont limitées à des emplacements fixes, ce qui réduit la connaissance sur leur variabilité spatiale. Afin de compléter cette information, un système mobile d’observation des aérosols (MAMS - Mobile Aerosol Monitoring System) a été mis au point pour explorer la variabilité verticale et spatiale des propriétés optiques et microphysiques des aérosols. MAMS est un « laboratoire » conçu sur un véhicule, se distinguant des autres plateformes transportables par sa capacité à effectuer des mesures au cours du mouvement. Equipé d'instruments de télédétection et in situ, MAMS a exploré la variabilité des aérosols au travers de campagnes mobiles sur routes dans diverses situations. Ce travail présente les études de variabilité et des événements remarquables observés en France et en Chine. L’une des variables aérosols recherchée est leur concentration massique et leur distribution verticale. Ce défi est relevé dans ce travail. Le système mobile offre une grande flexibilité, et un bon rapport coût-efficacité, car il permet de réagir rapidement en cas d’événements aérosols soudains, tels que des épisodes de pollution, poussières, incendies ou éruptions volcaniques, démontrant son utilité pour les réseaux d’observation recherche et opérationnels. Les applications de la plateforme d'exploration mobile pour les activités de validation des missions spatiales d’observation de la Terre et de l’atmosphère, pour l’évaluation des modèles et les campagnes d’intercomparaison d’instruments sont présentées. Des nouvelles perspectives, tant au niveau instrumental que méthodologique, sont introduites, laissant la place à des améliorations dans les observations mobiles des aérosols
Aerosols are a variable component of the atmosphere, having an important and complex role on air quality and climate. Most of the ground-based aerosol observations are limited to fixed locations, narrowing the knowledge on their spatial variability. In order to overcome this issue, a compact Mobile Aerosol Monitoring System (MAMS) has been developed to explore the vertical and spatial variability of aerosol optical and microphysical properties. MAMS is a vehicle-based laboratory, distinguished by other transportable platforms through its ability to perform measurements during the movement. Equipped with remote sensing and in situ instruments, MAMS explored the aerosol variability through mobile on-road campaigns in various situations. This work presents variability studies and remarkable events observed both in France and in China. One of the researched aerosol variables is their mass concentration and vertical distribution. This challenge is taken up in this work. The mobile system presents a great flexibility and is cost-effective, being able to respond quickly in case of sudden aerosol events such as pollution episodes, dust, fire or volcano outbreaks, proving to be a useful platform for aerosol observation and operational networks. Applications of the mobile exploratory platform for validation activities of space missions observing the Earth and the atmosphere, for models’ evaluation and for instrument intercomparison campaigns are presented. New perspectives, both on instrumental and methodology levels are introduced, leaving place for improvements in the mobile aerosol observations

En el presente estudio se analizan los efectos de los fotoperiodos ambientales de otoño e invierno y los fotoperiodos experimentales de 24, 12 y 0 horas de luz artificial sobre la calidad del semen de machos reproductores porcinos de raza Landrace.
El estudio se realizó sobre 30 machos postpuberales de 8 meses de edad y testados con el fin de comprobar la homogeneidad. Los machos fueron distribuidos aleatóriamente en los 3 grupos de luz artificial durante 3 meses. El tratamiento de 12 horas de luz artificial fue considerado como grupo control. Previamente al inicio de cada tratamiento, se sometió a cada grupo de machos al fotoperiodo ambiental correspondiente a la época del año; así, se caracterizó también la calidad seminal en otoño e invierno, manteniendo la temperatura constante.
La nave experimental que acogió a los machos tiene una superficie de 100 m2 y una altura de 3,5 m. Un pasillo central divide la nave en dos hileras de 5 y 6 celdas respectivamente. En una de las celdas pequeñas se instaló el maniquí y fue utilizada para las extracciones de semen. La iluminación artificial se consiguió con la instalación de 6 lámparas fluorescentes en el techo del pasillo central que proporcionaron una luz homogénea superior a 200 lux. Así mismo, la nave se mantuvo en todo momento a 21±1ºC y la humedad relativa osciló entre el 60-75%.
A todos los verracos se les proporcionó una dieta nutritiva y equilibrada y se les sometió a un regimen de extracciones de semen de 2 veces por semana, habiendo sido previamente entrenados en la monta del maniquí.
Les muestras de semen fueron recogidas según la técnica de la mano enguantada (Martín, 1982; Daza, 1992) y se analizaron los siguientes parámetros: el volumen y el pH seminales, la concentración, la vitalidad y la motilidad espermáticas, la resistencia acrosómica de los espermatozoides, la morfología espermàtica a partir de la frecuencia de los espermatozoides maduros, inmaduros y aberrantes, la producción testicular y el número de dosis seminales. Se analizó, además, bioquímicamente el plasma seminal al principio y al final de cada tratamiento experimental de luz artificial a partir de la concentración de proteína total, de la identificación de residuos fosforilados de proteína y del contenido de azúcares. También se determinaron los índices de fertilidad y prolificidad.
El volumen y el pH de los eyaculados se utilizaron como marcadores del estado funcional de las glándulas sexuales accesorias; la concentración espermàtica como un indicador de la actividad testicular (Pinart y col., 1999). La vitalidad y la motilidad espermáticas fueron estimadores del grado de diferenciación del espermatozoide tanto a nivel testicular como epididimario; la resistencia acrosómica fue utilizada para valorar el nivel de diferenciación de la membrana acrosómica durante la espermiogénesis y/o maduración epididimària (Briz i col., 1996; Pinart i col., 1999). Referente a la morfología espermática, los espermatozoides inmaduros fueron marcadores de anomalías en la maduración de éstos a lo largo del conducto epididimario y los espermatozoides aberrantes se utilizaron como marcadores de una diferenciación defectuosa a nivel de testículo (anomalías primarias) y a nivel de conducto epididimario (anomalías secundarias) (Briz i col., 1996). La concentración de proteína total se utilizó para valorar la integridad funcional de las membranas del espermatozoide y la actividad de las glándulas sexuales accesorias. La identificación de proteínas con residuos de tirosina fosforilados fue un estimador de la viabilidad celular y la actividad de las glándulas sexuales, y el contenido de azúcares como un indicador de la producción de las vesículas seminales.
La determinación del volumen y el pH de los eyaculados se realizó en las instalaciones de la granja a partir de semen fresco el mismo día de la extracción. El resto de parámetros se analizaron en el laboratorio durante las 48 horas posteriores a la extracción a partir de semen diluido en BTS (diluyente de Bestville) (Daza, 1992) y transportado y conservado a 15ºC. Las muestras fueron previamente filtradas con el fin de eliminar la tapioca.
El estudio estadístico de los resultados obtenidos se realizó a partir del análisis de la varianza (ANOVA) con un nivel de significación de =0,05.
En cuanto al estudio comparativo de los fotoperiodos ambientales estacionales se ha observado un incremento significativo del pH del eyaculado en los machos expuestos a otoño (P0,0001), mientras que el volumen seminal se mantiene en valores similares en ambos tratamientos (P=0,1650). La concentración espermàtica, la producción espermàtica y el número de dosis seminales que se pueden preparar a partir de un eyaculado se duplica en los verracos sometidos al fotoperiodo de primavera (P0,0001). La vitalidad y la motilidad espermáticas no experimentan cambios significativos entre tratamientos (P=0,3440 y P=0,9220, respectivamente). La resistencia osmótica de los acrosomas desciende únicamente en los machos expuestos a condiciones estacionales de otoño (P0,0001). En referencia a la morfología espermàtica aunque no se observan diferencias entre primavera y otoño (P0,05), sí se detecta un incremento de los porcentajes de espermatozoides inmaduros y aberrantes en ambos fotoperiodos estacionales, y en especial en los machos expuestos a condiciones fotoperiódicas de otoño. Según los resultados obtenidos en este estudio la calidad seminal de los verracos es inferior en el fotoperiodo de otoño debido a un descenso de la concentración y la producción espermáticas, un aumento del pH seminal, una disminución de la resistencia de la membrana acrosómica y a un incremento en la frecuencia de espermatozoides inmaduros y aberrantes. Parece ser, pues, que en el otoño tiene lugar la disminución de la producción testicular, cambios en la actividad de las glándulas sexuales accesorias y disfunciones en el proceso de diferenciación testicular y epididimària de los espermatozoides y especialmente del acrosoma.
En relación a los resultados obtenidos en el estudio de los diferentes fotoperiodos artificiales se observa que la iluminación continua provoca un aumento significativo del volumen del eyaculado en el primer y segundo mes de tratamiento (P0,0001), disminuyendo en el tercer mes. La oscuridad absoluta no modifica este parámetro (P0,05). En cuanto al pH seminal la iluminación continua provoca un incremento progresivo del valor del pH a lo largo del periodo experimental (P0,0001), mientras que la oscuridad absoluta tiene un efecto más irregular. La exposición de los machos a iluminación continua y a oscuridad absoluta se manifiesta en un descenso de la concentración y la producción espermáticas que se mantiene hasta el segundo mes de tratamiento (P0,0001), observándose un incremento en el tercer mes de exposición de los machos a oscuridad absoluta (P=0,1010). De todas maneras, este descenso es mas severo en los machos sometidos a iluminación continua ya que no presentan recuperación. La vitalidad y la motilidad espermáticas no se ven alteradas por la iluminación continua y la oscuridad absoluta, ni tampoco el contenido de los azúcares mayoritarios del plasma seminal (P0,005). La glucosa aparece como un azúcar minoritario y sí que presenta concentraciones inferiores en los tratamientos experimentales de luz continua y de oscuridad absoluta (P0,0001 y P=0,0002, respectivamente). La resistencia osmótica de los acrosomas desciende en ambos tratamientos artificiales extremos de luz continua y oscuridad total (P0,0001), aunque en los machos expuestos a iluminación continua se produce una recuperación a partir del segundo mes de tratamiento (P=0,4930). Dado que tampoco se han observado diferencias significativas en las concentraciones de proteína total (P0,05), es probable que las anomalías de la membrana acrosómica se originen durante el proceso de espermiogénesis y/o maduración epididimària. La exposición de los verracos a oscuridad absoluta no altera la morfología espermàtica de los eyaculados, aunque se observa un aumento de la frecuencia de espermatozoides con anomalías en la forma de la cola en el primer mes (P0,0001), y un aumento de la frecuencia de espermatozoides inmaduros con gota distal y de espermatozoides con anomalías en el número de colas en el tercer mes de experimentación (P=0,0030 y P0,0001). La luz continua, sin embargo, provoca un incremento de la frecuencia de espermatozoides inmaduros con gota distal (P0,0001) y de espermatozoides con anomalías en la forma de la cola (P=0,0040) ya en el primer mes. El fotoperiodo provoca un descenso de la fertilidad de los machos expuestos a oscuridad absoluta en el tercer mes de tratamiento (P0,0001) y un incremento de ésta en los machos sometidos a iluminación continua (P=0,0005). La prolificidad no se ve modificada por ambas condiciones extremas de luz artificial (P0,05). Así pues, los resultados obtenidos demuestran que el fotoperiodo afecta la actividad testicular, provoca alteraciones en la actividad de las glándulas sexuales accesorias, altera el proceso de expulsión de la gota citoplasmática y provoca anomalías en el proceso de diferenciación de la cola tanto a nivel testicular como epididimario, siendo los verracos expuestos a luz continua más sensibles a estos parámetros que los verracos sometidos a oscuridad absoluta. El fotoperiodo, sin embargo, no altera de forma esencial la integridad de las membranas del espermatozoide ni la capacidad fecundante de éste.
The present study analizes the effects of ambiental photoperiods of srping and autumn and of experimental photoperiods of 24, 12 and 0 hours lighting on the semen quality of Landrace boars.
For this purpouse 30 healthy postpuberal boars of 8 months of age were selected; the males chosed showed both facility to mount on the dummy and high semen quality. In order to determine the effects of light on seminal features. Boars were randomly distributed into 3 experimental groups throughout 3 months. The artificial photoperiod of 12 h was considered a control treatment. Previous to the initiation of each experimental treatment, the males were exposed in a natural photoperiod of autumn or spring for 15 days by maintaining the temperature constant.
In all groups the boars were kept in a experimental room of 100 m2 and 3,5 m high, divided into 2 files of 5 and 6 boxs, respectively. One box contained the dummy and was reserved for semen collections. The artificial lighting, upper the 200 lux, was obtained from 6 regulaly distributed lamps in the ceiling. The experimental room was always maintained in a temperature of 201ºC and an humidity of 60-75%.
Boars were fed with a nutritious diet, and subjected to a semen collection rythm of twice per week. Semen samples were obtained by the gloved hand method. For each semen sample the parameters analized were the ejaculate volume and pH, the sperm concentration, vitality and motility, the acrosomic resistance, the sperm morphology from the frequency of mature, immature, and aberrant spermatozoa, the testicular production and the number of dosis for each ejaculate. Moreover, samples from the day 30th and 90th of each experimental treatment were processed for biochemical analysis in order to determine protein concentracion and identificate the tyrosine phosphorilated residues and to determine fructose, sorbitol and glucose concentrations; and, also they were processed to value fertility and prolificity indexs.
The volume and pH of the ejaculates were measured from the sperm fraction the same day of the extractions. The other seminal parameters were analized in the 48 hours after the obtention of the samples from the filtration and dilution of the sperm fraccion. Statistical comparisons were made using the analysis of variance ANOVA at a significance level of =0,05.
The comparative study of natural photoperiods indicated that pH was significantly higher in males under autumn conditions, while seminal volume did not differ. Sperm concentration, sperm production and number of seminal dosis were higher in males exposed to spring conditions. Sperm vitality and sperm motility was similar in males exposed to both spring and autumn conditions, whereas osmotic resistance of acrosomes was lower in autumn photoperiod. The sperm morphology did not differed between treatments, but in males exposed to autumn photoperiod an increase of the frequency of inmature and aberrant spermatozoa was found.
These results show that the sperm quality of boars was lower in autumn photoperiod than in spring photoperiod due to low sperm concentration and sperm production, a high pH value of semen, a low osmotic resistance of acrosome and a high frequency of inmature and aberrant spermatozoa. Thus, autumn conditions induces a decrease in testicular production, and dysfunctions in both testicular and epididymal differentiation of spermatozoa
The comparative study of artificial photoperiods showed that 24 h lighting resulted in an increase of seminal volume in the first and second months of treatment, whereas 0 h lighting did not affect this parameter. The pH of semen increased progressively throughout the 24h treatment, whereas the 0h treatment showed an irregular effect on this parameter. Both experimental conditions produced a decrease in sperm concentration and sperm production until the second month; this decrease was more severe in males exposed to 24 h lighting. Neither the sperm vitality and motility nor the fructose and sorbitol concentration were affected by experimental condictions. In our study, glucose appeared as a minoritary sugar that was affected by artificial photoperiod. The effect of artificial photperiod on osmotic resistance of acrosome was more severe in darkness than in continuous lighting. The exposure of boars to darkness and continuous lighting did not alter the sperm morphology; however an increase of the frequency of inmature spermatozoa with distal droplet and aberrant spermatozoa with tail anomalies. The total protein concentration and the content of phophorilated tyrosine residues showed normal values throughout the experimental treatments. Fertility and prolificity were not affected by experimental conditions; however in males exposed to darkness a decrease in fertility was found in the third month of threatment.
Data obtained indicate that photoperiod affects the semen quality of boars by decreasing the sperm concentration and sperm production, the osmotic resistance of acrosome and the glucose concentration, and by increasing the frequencies of inmature and tail aberrant spermatozoa; these alterations were more severe in those males exposed to 24h lighting thant in males exposed to 0h lighting. However, despite these anomalies both the fecundity and prolificity are not disturbed. Lack of abnormalitites in fecundity and fertility are correlated with the normal content of both sugar and proteins in seminal plasma of boars.

碩士
國立屏東科技大學
動物科學與畜產系所
100
Heterologous insemination (HI), also known as heterospermic inseminations, is a common practise to inseminate sows with extended semen made of ejaculates from 2 or more boars. HI may increase sows’ conception rates by ‘risk diversification’, especially when boar’s fertility is not certain. Because of this , HI has becomed popular in Taiwan. However, it is still not clear that ejaculate mixing would affect quality of the extended semen during storage. This study was conducted to compare daily change in semen traits for three days among various combinations of heterospermic semen. In experiment 1 (Exp 1), three ejaculates each from 29 Duroc boars were collected and there motility (MOT) and hypoosmotic swelling test (HOST) were determined. Boars were classified into one of three groups by cluster analysis, including SH (single ejaculate with high MOT and HOS), SM (low MOT but high HOST), and SL (high MOT but low HOST). In Exp 2, each four combinations of heterospermic semen samples were made, including HH, HM, HL, ML, and MM, by either mixing equal sperm number (MEN-) or equal ejaculate volume (MEV-). These combined or noncombined semen sample were all extended into a typical dose (5×108 total sperm in 80 mL) before stored at 16℃. Sperm traits were measured daily for 3 days. In Experiment 3, ejaculates from same boars were mixed at a ratio of 1:2 or 2:1. The mixed and unmixed samples were prepared and sperm traits were examined as described in Exp 2. Data were arc-sine transformed and then analyzed using ANOVA, orthogonal contrast, and Duncan multiple range test. Among MEN groups in Exp 2, MEV-HL group exhibited a higher MOT than SH and SL (56.2 ± 5.2% vs 66.4 ± 2.3 and 64.8 ± 3.7%). SL group had a lower HOST than SH, SM and any other combined groups. MEN-MM group had lower acrosome integrity than all uncombined groups and the rest of combined groups. Among MEV groups in Exp 2, MEV-HH and MEV-HM had a higher MOT than SM group (69.7 ± 3.2, 70.0 ± 2.9% vs 60.9 ± 2.4%). All the heterospermic groups had a higher HOST than SL group. In addition, SH (41.9 ± 1.5%) and SM (43.1 ± 1.3%) groups both had higher HOST than MEV-HL (36.8 ± 2.9%) (P < 0.05). In Exp 3, M-HL12 had a higher MOT than M-HL21 (62.8 ± 4.8% vs 53.1 ± 5.3%). Groups SH and SM had higher HOST than M-HL12 (37.2 ± 2.6%), M-ML12 (34.7 ± 2.6%), M-HL21 (34.5 ± 3.1%) and M-ML21 (36.1 ± 2.5%). SM group also exhibits a higher HOST than M-HM12 (41.4 ± 2.1%). M-HM21 group had a higher acrosome integrity than M-HM12 (69.8 ± 3.5% vs 64.4 ± 4.0%), while acrosome integrity in SH group (69.2 ± 2.8%) was higher than groups M-HM12 and M-HL21 (64.4 ± 4.0%, 64.8 ± 3.5%), and M-HM21 higher than SM (65.1 ± 3.8%). These results suggest that ejaculates mixed with equal volume tends to have higher while ejaculates mixed with equal number tend to have a lower motility than unmixed ones. Extended semen made from single ejaculate also had higher motility than mixed group, no matter what the ratios of sperm numbers were. Semen made from equal number sperm may have a higher HOST than those mixed with equal volumes or the unmixed groups, and the unmixed group have a higher HOST than those mixed with various ratios. It is concluded that ejaculate mixing may exhibit dissimilar effects on measurements of motility and HOST.

碩士
國立宜蘭大學
生物技術與動物科學系動物科學碩士班
102
Liquid and frozen methods are usually used to apply in boar semen preservation. The principal semen preservation is liquid method in pig breeding farm. However the number of semen dosages of collected and diluted the boar semen is usually greater than the number of mated sows. It wastes the excellent boar semen. If we could cryopreserve the liquid boar semen, we would increase the application of the excellent boars. The objective of this study was to evaluate the effect of Trolox in diluent boar semen for 2 and 3 days storage on the sperm quality and in vitro fertility of cryopreserved boar diluent semen. In experiment 1, the fresh boar semen was mixed with SafeCell diluent containing 200 μM Trolox in a 50% ratio, and stored at 16 to 18 ˚C in a refrigerator for 2 and 3 days. Before and after cryopreservation, the sperm motility, acrosomal integrity, viability and mitochondrial activity of the stored semen were analyzed using phase contrast microscopy and flow cytometry. The thawed semen was examined during 4 hours (0, 2 and 4 hours) post-thawed. The results indicated that the sperm quality parameters including motility, acrosomal integrity, viability and mitochondrial activity of liquid stored semen with Trolox after 2 d and 3 d storage were no significant difference compared with the control. The acrosomal integrity of cryopreserved diluent semen with Trolox for 2 d storage was 37.8% that was significantly better than the control (28.1%) (P < 0.05). At post-thawed 2 hours, the motility of sperm in diluent semen with Trolox for 2 d storage was 46% that was significantly higher than the same treatment for 3 d storage (40%) (P < 0.05). At post-thawed 4 hour, all quality parameters of the 4 groups were not different significantly.In experiment 2, fresh boar semen was mixed with SafeCell or BTS diluents containing 200 μM Trolox in a 50% ratio, and stored at 16 to 18 ˚C in a refrigerator for 3 d. Before cryopreservation, the free radical scavenging of diluent semen sored at 0 and 3 days was analyzed by DPPH antioxidant assay. Before and after cryopreservation, the sperm motility, acrosomal integrity, viability and mitochondrial activity of the cryopreserved semen were analyzed by phase contrast microscopy and flow cytometry. Finally, the fertility of the frozen semen was evaluated by in vitro fertilization of porcine oocytes. The results showed that the free radical scavenging among all groups was not significantly different before semen cryopreserved. The quality parameters of thawed semen, including motility, acrosomal integrity, viability and mitochondrial activity did not differ significantly between SafeCell and BTS diluent semen with or without Trolox. However, the incubated time (0, 2 and 4 hours) of post thawed semen affected all quality parameters. The longer time of incubation decreased the sperm quality. The diluent semen supplemented with Trolox did not improve the sperm quality of cryopreserved semen. Finally, there was no difference of penetration rate and polyspermy rate in IVF experiments among the 4 groups.In conclusion, the Trolox in diluent semen could improve the integrity of sperm acrosome, and extension the time of liquid stored semen decreased the sperm motility of frozen-thawed semen. All parameters and fertility of BTS and SafeCell diluent semen with or without Trolox that stored 3 days and cryopreserved did not differ significantly after been thawed. We also suggested that the shorter time diluent (BTS) could be used to dilute boar semen for cryopreservation, but the longer time diluent (SafeCell) maintained the better sperm quality of post-thaw semen.

碩士
國立屏東科技大學
動物科學與畜產系所
105
Cryopreservation process produces significant changes in mammalian spermatozoa with the subsequent decrease in its fertility potential. Different factors are involved in the alteration of sperm during the cryopreservation: the cryoprotectant used and chilling, freezing and thawed processes employed. Therefore, a large number of extenders have been studied in order to preserve the canine semen quality during the cryopreservation. This study was investigated to explore the effect of glycerol (G), low density lipoprotein (LDL) and trehalose concentration on post-thawing canine semen quality. Semen were collected from three mature Maltese canine (2 and 5 years of age)；The aim of this study is to evaluate the semen quality and is divided into motility (%), progressive (%), viability (%), acrosome integrity (%), mitochondrial integrity (%), and DNA integrity. Results showed that the final glycerol concentration of 5% of the cryoprotectant is significantly higher (P <0.05) than other extender is 3%, 4%, 6% and 7% glycerol concentration in 11% lactose with 20% egg yolk basic extender. When the extender added 11% LDL, canine semen quality (P < 0.05) is significant higher than 10%, 12%, 13% and control group (20% egg yolk) in 11% lactose with 5% glycerol-based extender. Based on the 11% LDL with 5% glycerol based extender, the final optimal trehalose concentration of canine semen cryoprotectant is 350 mM which is better than 300 mM and the control group (11% lactose). In conclusion, the best concentration of glycerol, LDL and trehalose for freezing canine semen are 5 %, 11% and 350 mM. Keywords: Canine, Cryoprotectation, low density lipoprotein, Glycerol, Semen quality, Trehalose

碩士
南華大學
自然醫學研究所
94
This study is intended to assess the purpose of employing Semen Zizyphi Spinosae and Semen Biotae, two kinds of traditional Chinese medicine (TCM), to improve the sleep quality of insomniac. For the purpose of conducting this assessment, 16 patients ( 2 males and 14 females, average age of 50.3 ± 13.4 ) diagnosed by a TCM doctor as yin-xu (passive-weak) insomnia were prescribed of concentrated granules of Semen Zizyphi Spinosae and Semen Biotae,（2 grams each） to be taken orally at 8 p.m. every night for a period of four weeks. The heart rate variabilities (HRV) of patients were measured five times each, prior and posterior to taking the granules. In the meantime, the Pittsburgh Sleep Quality Index (PSQI) would be employed and assessed by the patients themselves and their sleep quality would be further assessed by the TCM doctor. As the results, indicate the PSQI felt from prior 13.3 ± 3.1 points to posterior 8.4 ± 4.9 points (p＜0.05); and the TCM doctor assessment also declined from prior 12.6 ± 5.4 points to posterior 2.2 ± 3.7 points (p＜0.05). These results clearly indicate that Semen Zizyphi Spinosae and Semen Biotae are helpful to improve the sleep quality of yin-xu insomniacs. Since no any critical variances were observed with the HRV measurement, Semen Zizyphi Spinosae and Semen Biotae did not appear to impose any significant effects on the patients'' cardio-autonomic nervous system in the prescribed duration. It is concluded that Semen Zizyphi Spinosae and Semen Biotae can be employed as a choice of complementary and alternative medicine (CAM) to improve the symptoms of insomnia.