A predição do desempenho de touros na produção in vitro de embriões (PIVE), pela avaliação seminal, não está plenamente estabelecida. Com este estudo, objetivou-se caracterizar as diferenças nos atributos espermáticos de touros com baixo e alto desempenho, baseado nas taxas de clivagem e de blastocisto, visando estabelecer métodos preditivos de maior eficiência na PIVE. A partir do histórico de três anos da empresa In Vitro Brasil®, foram selecionados touros que haviam resultado nas menores e maiores taxas de clivagem e de blastocisto, estabelecendo-se 4 grupos experimentais: baixa taxa de clivagem (BC, n=5), alta taxa de clivagem (AC, n=5), baixa taxa de blastocisto (BB, n=5) e alta taxa de blastocisto (AB, n=5). Foram adquiridas palhetas de sêmen congelado dos touros de cada grupo, nas quais foram avaliados, antes (Pré-Percoll®) e após centrifugação em gradiente de Percoll® (Pós-Percoll®), os atributos espermáticos: cinética, em sistema CASA; integridade de membranas plasmática e acrossomal (FITC-PSA/PI), potencial de membrana mitocondrial (JC-1), status oxidativo (DHE, CellROXTM green/PI, MitoSox red), apoptose (FITC-FMK-VAD/PI) e DNA (SCSA), por citometria de fluxo; morfologia; atividade mitocondrial (DAB); capacitação (CTC); teste funcional e TBARS induzido. Diferenças entre grupos (BC vs. AC e BB vs. AB) foram estimadas pelo teste t. Para todas as análises estatísticas foi considerado nível de significância de 5%. Na ausência de interações significativas, o efeito dos grupos foi analisado ao fundir os momentos Pré e Pós-Percoll®. Os grupos AC e AB apresentaram valores superiores de VAP, VCL, ALH e DNC e valores inferiores de BCF, STR, LIN e WOB, cinética indicativa de hiperativação. Adicionalmente, estes grupos apresentaram indícios de menor ocorrência de estresse oxidativo, evidenciados pelo percentual de espermatozoides com membrana lesada e estresse oxidativo (CellROXTM green/PI) no grupo AC e nível de DHE no grupo AB. Para os demais atributos estudados, não se obteve diferença entre os grupos. Por meio de análise de regressão logística multivariada, constatou-se que a taxa de clivagem sofre efeito de: VCL, BCF e percentual de espermatozoides com integridade de membrana e sem estresse oxidativo; já a taxa de blastocisto, sofre efeito de: defeitos menores, VSL, DABIII e DHE, os quais foram inclusos nos respectivos modelos preditivos. As taxas obtidas com o modelo preditivo foram comparadas às taxas reais por análise de correlação de Pearson, que resultaram em intensidade moderada tanto para clivagem (r=0,56) quanto para blastocisto (r=0,44). Subpopulações espermáticas com diferentes perfis de cinética em CASA, foram determinadas em três etapas estatísticas segundo Núñez-Martínez et al. (2006). Foram detectadas 4 subpopulações: 1-Rápidos e progressivos; 2- Hiperativados; 3- Lentos e sinuosos; 4- Lentos e progressivos. Touros com alto desempenho (AC e AB) apresentaram maior percentual de subpopulação 2, Pré-Percoll®, e das subpopulações 1 e 2, Pós-Percoll®. Com isto, demonstra-se ser possível diferenciar touros de baixo e alto desempenho na PIVE, pela avaliação dos atributos espermáticos, com o estabelecimento de modelos preditivos para as taxas de clivagem e de blastocisto. Ademais, o estudo das subpopulações espermáticas constitui-se no método mais efetivo para a predição do desempenho in vitro. / The prediction of bull performance on the in vitro production (IVP) of bovine embryos, by the seminal evaluation, is not fully established. Thus, the aim of this study was to characterize the differences in the sperm features of bulls with low and high performance, based on cleavage and blastocyst rates, in order to establish predictive methods for a greater efficiency in the IVP. Based on the results obtained in three years from the company In Vitro Brasil®, bulls were selected by the lowest and highest cleavage and blastocyst rates, establishing 4 experimental groups: low cleavage rate (BC, n=5), high cleavage rate (AC, n=5), low blastocyst rate (BB, n=5) and high blastocyst rate (AB, n=5). Frozen semen were obtained from the bulls of each group, in which were evaluated, before (Pre-Percoll®) and after Percoll® gradient centrifugation (Post-Percoll®), the sperm features: kinetics, in CASA; integrity of plasma and acrosomal membranes (FITC-PSA/PI), mitochondrial membrane potential (JC-1), oxidative status (DHE, CellROXTM green/PI, MitoSox red), apoptosis (FITC-FMK-VAD/PI) and DNA (SCSA), by flow cytometry; morphology; mitochondrial activity (DAB); capacitation (CTC); functional test and induced TBARS. Differences between groups (BC vs. AC and BB vs. AB) were estimated by t test. A significance level of 5% was considered for all statistical analysis. In the absence of significant interactions, the effect of the groups was analyzed by merging the moments Pre and Post-Percoll®. The AC and AB groups presented higher values of VAP, VCL, ALH and DNC and lower values of BCF, STR, LIN and WOB, kinetics indicative of hyperactivation. In addition, these groups showed less evidence of oxidative stress, observed by the percentage of spermatozoa with damaged membrane and oxidative stress (CellROXTM green/PI) in the AC group and level of DHE in the AB group. For the others sperm features, there was no difference between groups. By multivariate logistic regression analysis, it was verified that the cleavage rate is affected by: VSL, BCF and percentage of spermatozoa with membrane integrity and without oxidative stress; and the blastocyst rate, is affected by: minor defects, VSL, DABIII and DHE, which were included in the respective predictive models. The rates obtained with the predictive model were compared to the real rates by Pearson correlation analysis, which resulted in moderate intensity for both cleavage (r=0.56) and blastocyst (r=0.44). Spermatic subpopulations with different kinetic profiles at CASA were determined in three statistical steps according to Núñez-Martínez et al. (2006). Four subpopulations were detected: 1-Fast and progressive; 2- Hyperactivated; 3- Slow and sinuous; 4- Slow and progressive. Bulls with high performance (AC and AB) presented higher percentage of subpopulation 2, Pre-Percoll®, and subpopulations 1 and 2, Post-Percoll®. In summary, it is possible to differentiate low and high-performance bulls at IVP by evaluating the sperm features, with the establishment of predictive models for the cleavage and blastocyst rates. Furthermore, the study of sperm subpopulations is the most effective method for predicting in vitro performance.

El semen de verracos aparentemente sanos provenientes de granjas de crianza tecnificada y semitecnificada fueron evaluados bacteriológicamente para investigar la cantidad de microorganismos presentes. Se analizó 60 muestras de semen extraídas de 30 verracos, procedentes de tres granjas porcinas de Lima, dos granjas de crianza tecnificada y una semitecnificada. Cada muestra de semen fue sembrada en medios de cultivo y las muestras que tuvieron desarrollo bacteriano, fueron identificados bioquímicamente. En el 73% (11/15) de muestras de granjas tecnificadas se encontró crecimiento bacteriano y los gérmenes aislados según su frecuencia fueron Pseudomona aeruginosa, Citrobacter spp, Proteus vulgarís, Escherichia coli, Staphylococcus aureus, Bacillus subtilis y Micrococcus spp.. En el 100% (15/15) de las muestras de granjas semitecnificadas hubo crecimiento bacteriano, encontrándose Pseudomona aeruginosa con mayor incidencia y le siguieron Escherichia coli, Proteus vulgarís, Staphylococcus aureus, Bacillus subtilis, Klebsieila spp. y Citrobacter spp. En las granjas tecnificadas, el 13% (2/15) de reproductores sobrepasaron el límite de unidades formadoras de colonias por mililitro de semen (UFC/ml) establecido por la Oficina Internacional de Epizootias (OIE) (no >5103 UFC/ml) y en ia semitecnificada el 60% (9/15) superaron estos límites. Existen muchos factores que pueden actuar negativamente sobre la calidad espermática dei semental provocando ciertas alteraciones cualitativas y cuantitativas que van a repercutir a corto plazo en una disminución de la eficiencia reproductiva del animal. / Tesis

The purpose of this study was to evaluate the use of female selected sex-sorted semen and to determine the association of variables on the success of Western Kentucky University’s Dairy Farm. Official breeding and calving records (n=144) were used to determine the relation of lactation number, breeding season, breeding number, breeding year and semen type on pregnancy results, sex of offspring, and the mortality of the offspring. Previous research has shown pregnancy results can be influenced by lactation number, breeding season, number of breedings and semen type. Results from this study show that pregnancy results were not associated with lactation number (P=0.21), breeding year (P=0.22), breeding number (P=0.52) or semen type (P=0.99). Breeding season was associated with pregnancy results (P=0.04). Lactation number (P=0.40), breeding season (P=0.20) or breeding number (P=0.12) did not influence the sex of the offspring. The year of breeding and semen type (conventional or sexed) had a significant or close to significant effect on the sex of the offspring (P=0.01) and (P=0.06). The mortality of offspring was not associated with lactation number (P=0.46), breeding season (P=0.94), breeding year (P=0.76), breeding number (P=0.40) or semen type (P=0.49).

Typically, semen testing involves presumptive and confirmatory tests to determine the region in which a semen stain has been deposited prior to initiating DNA analysis. However, previous research showed that the soluble components of seminal fluid, but not spermatozoa, migrated from their original location on cotton cloth upon exposure to porcine decomposition fluids and rainfall/dew6. This indicates that preliminary testing and detection techniques may result in areas being sampled that will not yield a successful DNA profile. The present study assesses how various amounts of water or blood affect migration patterns of seminal fluid components using traditional serological screening methods as well as DNA analysis. The effects of exposing a semen stain to water over the course of several days are also investigated. The final component of the study evaluates whether the presence of acid phosphatase (AP) Spot reagent had any detrimental effects on subsequent antigen P30 (P30) testing, Kernechtrot Picroindigocarmine (KPIC) sperm staining or DNA analysis. Neat semen was deposited onto swatches from cotton sheets and allowed to dry before being sprayed with 2 mL, 5 mL, or 10 mL of water or blood. The swatches were allowed to dry while lying flat, at 45°, or at 90°. Three of the swatches were sprayed directly with AP Spot reagent to determine any potential interference with subsequent P30 and DNA testing. After the water or blood was dry, the swatches were viewed with an alternate light source (ALS) at 450 nm using orange barrier filter goggles. Three-millimeter fabric punches were collected from each swatch in at least thirteen locations (one from the center of the stain and four at 1 cm, 4 cm, and 7 cm from the perimeter of the stain in multiple directions), and were extracted for two hours prior to testing for the presence of P30. Additional fabric punches were collected from each P30 positive location to be used for DNA analysis. AP testing showed positive results beyond the original semen stain with an average distance of 1-3 cm from the perimeter of the original region of deposition (ORD) for all swatches except those moistened with blood. AP mapping was performed on the swatches moistened with blood and negative results were obtained. Positive P30 results were obtained for all swatches with an average distance of 1-3 cm from the ORD. The angle at which the swatch was positioned influenced the direction(s) that the soluble components migrated; however the amount of water (or blood) the swatch was exposed to had a much greater effect on the distance of migration. Microscopic examination of slides made from the extracts of each fabric punch revealed minimal spermatozoa migration for all swatches; the majority of the samples outside of the ORD showed no spermatozoa, although a few showed a single sperm cell. These findings demonstrate that the soluble components of semen stains that often aid in detection migrated when exposed to moisture, while sperm cells containing genetic material largely remained in their original location. The DNA analysis results confirmed the lack of spermatozoa migration. Full DNA profiles were obtained from within the ORD of the flat and 90° swatches. The samples from outside of the ORD produced either partial profiles (maximum dropout rate of 97%) or no profile. If case circumstances suggest that evidence has been exposed to water, multiple regions should be tested in order to maximize the possibility of identifying semen and obtaining a DNA profile. AP Spot reagent was not found to have detrimental effects on P30 testing, sperm staining or DNA analysis. Therefore, direct application of AP Spot reagent could be used for larger pieces of evidence where the location of a stain is unknown. This would eliminate the careful documentation needed for chemical mapping and the reliance on the transfer of acid phosphatase from one substrate to another.

In the stallion, inconsistent sexual behavior and variable semen quality are common. This reproductive variability has been attributed to differences in circulating hormone concentrations. In order to further examine this relationship, 7 miniature stallions were observed for sexual behavior and semen characteristics. Blood was also drawn from each stallion 15 min before mating (time -15), immediately following ejaculation (time 0) and at times following ejaculation (times +15, +30, and +60). Plasma was later analyzed for concentrations of testosterone (T), estrone sulphate (ES) and cortisol. Semen was evaluated for volume, sperm concentration and progressive motility. Sexual behavior was quantified by assigning a libido score to each stallion, recording reaction time and the number of jumps required for ejaculation. Upon statistical analysis, data revealed both ES and cortisol increased at the time of semen collection (P < 0.05), while T did not. Regression analysis revealed that ES and the ratio of ES to T at times -15, +30, and +60 were negatively correlated to libido scores. Additionally, a positive relationship was found between ES at times -15 and +60 and reaction time, as well as between cortisol at times -15, 0, and +15 and libido scores. No relationship was observed between T and sexual behavior. However, T at time -15 was positively correlated to progressive motility, and the ratio of ES/T at time -15 was negatively correlated to progressive motility. No other association was detected between ejaculate parameters and hormone concentrations. These results not only serve to enhance understanding of stallion hormone profiles, but also provide further insight into the hormonal control of sexual behavior and sperm production. This knowledge can be used to generate improved management techniques for stallions that are inconsistent in sexual behavior and sperm output.

Determination of an extender protocol which will enhance the viability of frozenthawed bovine spermatozoa will allow producers to obtain higher conception rates due to the increased survival rate of the spermatozoa. Ejaculates of six Brangus bulls (age=18 months) were evaluated for spermatozoal motility, acrosomal integrity, and morphological characteristics (collectively called spermatozoal viability) in two experiments to test our hypotheses that (1) the treatment combination of a 4 hr cooling duration and a 2 hr equilibration with glycerol will result in optimum spermatozoal characteristics after freezing and thawing and (2) rank of three selected extenders relative to their effects on spermatozoal viability after freezing and thawing will be egg yolk-citrate (EC), egg yolk-tris (IMV), and skim milk (milk). In experiment 1, an ejaculate from each bull was partially extended and cooled to 4 ºC for either 2 or 4 hr and then allowed to equilibrate with the glycerolated extender for 2, 4, or 6 hr. Spermatozoal viability was assessed at 0, 3, 6, and 9 hr after thawing. In experiment 1, 4 hr of cooling resulted in a higher percentage of motile spermatozoa than did 2 hr of cooling. The 2 hr equilibration with glycerol yielded lower percentages of motile spermatozoa, acrosomal integrity, and morphologically normal spermatozoa than 4 and 6 hr equilibration durations with glycerol. In experiment 2, we observed a decrease in spermatozoal viability for all three extenders upon freezing and thawing. Viability of frozen-thawed spermatozoa extended in the milk was reduced for all incubation durations, and the IMV extender had a higher percentage of motile spermatozoa than the EC extender at 6 hr of incubation. A higher percentage of intact acrosomes was observed with the IMV extender; however, the EC extender had a higher percentage of morphologically normal spermatozoa than the IMV extender. Our results indicate that at cooling duration of 4 hr and a 4 hr equilibration with glycerol provide the highest level of spermatozoal viability post-thaw of the treatments evaluated and that the IMV extender enhances the percentage of spermatozoa with an intact acrosome for frozenthawed spermatozoa over the EC and skim milk extenders.

El objetivo del presente estudio fue determinar el tiempo máximo para recuperar y criopreservar espermatozoides obtenidos de la cola del epidídimo de caninos post orquiectomía. Los testículos/epidídimos fueron obtenidos después de la orquiectomía de 20 caninos adultos aparentemente saludables con edades entre 1 y 8 años. Los testículos/epidídimos fueron colocados en cloruro de sodio al 0.9% y almacenados a 5ºC durante 0, 24, 48 y 72 horas. Los espermatozoides fueron recuperados al cortar la cola del epidídimo suspendido en dilutor Tris-citrato-fructosa. Para el proceso de criopreservación, a la muestra diluida (espermatozoides + dilutor) se le añadió yema de huevo (20%) y glicerol (5%). La nueva dilución fue envasada en pajillas de 0.5ml, las cuales fueron sometidas a una curva de enfriamiento para luego ser colocadas en nitrógeno líquido. Los parámetros evaluados fueron: Motilidad total, motilidad progresiva e integridad funcional de membrana; parámetros que fueron analizados antes y después de la criopreservación. Adicionalmente se obtuvieron datos sobre morfología y concentración, evaluadas sólo antes de la criopreservación. Todos los parámetros evaluados disminuyeron gradualmente a medida que aumentó el tiempo de almacenamiento, los cuales al ser evaluados a las 48 horas de almacenamiento, antes y después del proceso de criopreservación, no variaron significativamente con respecto a los evaluados a las 0 horas. Sin embargo, cabe resaltar que todos los valores obtenidos después del proceso de criopreservación fueron marcadamente inferiores a los obtenidos antes del proceso. / The aim of this study was to determine the maximum time to recover and cryopreserve sperm from the tail of the epididymis of canine post-orchiectomy. The testes/epididymides were obtained by orchiectomy of 20 apparently healthy adult dogs between 1 to 8 years old. The testes/epididymides were placed in sodium chloride 0.9% and stored at 5 ° C for 0, 24, 48 and 72 hours. Sperm were recovered by cutting the tail of the epididymis dilutor suspended in Tris-citrate-fructose. For the cryopreservation process, the diluted sample (sperm + dilutor) was added egg yolk (20%) and glycerol (5%). The new dilution was packaged in 0.5ml straws, which were subjected to a cooling curve to be later placed in liquid nitrogen. The parameters evaluated were: Total motility, progressive motility and membrane functional integrity; parameters were analyzed before and after cryopreservation. Additionally, data were obtained on morphology and concentration, evaluated just before the cryopreservation. All evaluated parameters decreased gradually when the storage time increased. Such parameters to be evaluated at 48 hours of storage, before and after the cryopreservation process, did not differ significantly from those evaluated at 0 hours. However, it is noted that all values obtained after the cryopreservation process were markedly lower than those obtained before process.

El objetivo del estudio fue evaluar el efecto de cuatro combinaciones de dos agentes crioprotectores permeantes más dos no permeantes sobre la calidad del semen ovino post-descongelamiento. Para esto, primero (Experimento 1) se seleccionó, entre tres dilutores (A, B y C), el más adecuado para ser usado en la segunda parte (Experimento 2), en la cual se evaluaron las siguientes combinaciones de agentes crioprotectores permeantes y no permeantes: 1)Glicerol - Trehalosa, 2)Glicerol - Sacarosa, 3)Etilenglicol - Trehalosa y 4)Etilenglicol - Sacarosa sobre la calidad del semen ovino post-descongelamiento. Para tal efecto, se realizaron 6 repeticiones para el Experimento 1 y 8 repeticiones para el Experimento 2. Cada repetición constó de 4 eyaculados cada una. En el experimento 1 se encontró que el Dilutor A mantenía mejor la motilidad progresiva y viabilidad e integridad acrosomal post-descongelamiento (69% y 63%) en comparación con el Dilutor B (37% y 35%) y el Dilutor C (23% y 23%). Por lo cual, el Dilutor A fue empleado en el experimento 2. En el experimento 2 se encontró que la motilidad progresiva, la viabilidad e integridad acrosomal, la termoresistencia y la integridad de membrana plasmática post-descongelamiento fue mejor en los grupos con glicerol-sacarosa (64%; 56%; 40% y 63%) y glicerol-trehalosa (64%; 58%; 38% y 60%) en comparación con los grupos con etilenglicol-sacarosa (48%; 42%; 25% y 37%) y etilenglicol-trehalosa (44%; 38%; 27% y 40%). Lo cual demuestra que el glicerol es un mejor crioprotector permeante en comparacion con el etilenglicol. Sin embargo, no existe diferencia en el uso de sacarosa o trehalosa. Se concluye que un dilutor con las características del dilutor A, utilizando glicerol más trehalosa o sacarosa constituye una buena alternativa para la criopreservación de semen ovino. / The objective of the study was to evaluate the effect of four combinations of two permeant and two non permeant cryoprotectant agents on the quality of post thaw ram semen. In Experiment 1, three extender were evaluated (A, B, and C) in order to have th ebetter one for the next step. In Experiment 2, different combinations of cryoprotectant agents were evaluated as follow: 1) Glycerol–Trehalose, 2) Glycerol–Sucrose, 3) Ethyleneglycol-Trehalose, and 4) Ethyleneglycol–Sucrose. In this way, 6 assays for Experiment 1 and 8 assays for the Experiment 2 were carry out. In each assay, 4 ejaculated were mixed to work as an original sample. Percentages of motility, viability and acrosomal integrity in extender A were higher (69% and 63%) than extender B (37% and 35%) and extender C (23% and 23%). Therefore, extender A was used for experiment 2. In the experiment 2, motility, viability and acrosomal integrity, thermoresistance, and plasmatic membrane integrity were higher in groups Glycerol-sucrose (64%; 56%; 40%, and 63%) and Glycerol-trehalose (64%; 58%; 38%, and 60%) in comparison with groups Ethileneglycol-sucrose (48%; 42%; 25% and 37%) and Ethileneglycol-trehalose (44%; 38%; 27% and 40%). However, there is not significative differences between sucrose or trehalose. In conclusion, an extender with characteristics of extender A, it means, using glycerol plus trehalose or sucrose constitute a good alternative for cryopreservation of ram semen.

Shortly after returning to Moscow in 1936, Prokofiev composed his first Soviet opera, Semyon Kotko (Opus 81). The libretto was taken from Valentin Kataev's novel I am a Son of the Working People, a tale of revolution and war in a small Ukrainian village and one that adheres to the tenets of Socialist Realism. Kataev encouraged Prokofiev to set this text in a highly conservative song style. Prokofiev was also influenced in the project by Vsevolod Meyerhold, an innovative artist who advocated using continuous declamation as a means of achieving "dramatic truth" in music. / This essay examines the extent to which Semyon Kotko can be considered a conformist opera. Part One is a survey of Socialist Realism and its manifestation in Soviet literature and music during the 1930's; Parts Two and Three examine the text and music of Semyon Kotko as representative of the doctrine. Consideration is given throughout the study to the opposing influences of Kataev and Meyerhold on Prokofiev, and to the political events surrounding the opera's composition.

No description available.