Factors affecting fertility in frozen-thawed boar sperm

Frozen-thawed boar sperm holds the potential to have an impact on the future of the swine industry. Utilization of this technology could improve a swine producer’s ability to access top-tier genetics from around the world, to improve efficiency, profitability, and the quality of product to meet consumer demands. Effective application of frozen-thawed sperm can help reduce the potential risk associated with devastating economic loss due to the spread of disease. Frozen storage of boar sperm also provides a safeguard in the event of disease outbreaks, as genetic material from paternal lines can be preserved and banked for repopulation purposes. Historically these benefits have been masked by reduction in fertility measures such as litter size. The reduced fertility results from the damage sustained by the sperm cell during cryopreservation. However, increased understanding of this damage has lead to improved cryopreservation methods, ultimately increasing post-thaw viability and fertility. Enhancements in breeding technology have also resulted in a better understanding of the AI methods required to achieve acceptable farrowing rates and litter size. Fertility following AI with frozen-thawed sperm is approaching that of liquid stored sperm, and producers may soon reap the benefits of this technology. This thesis will outline the current swine industry, opportunities for utilizing frozen-thawed sperm, the main components of sperm, why they are susceptible to damage, and current freezing and breeding practices. Objective 1 was to develop a cryopreservation protocol for our lab that resulted in consistent post-thaw motility ( ≥ 40%) that would eventually be used by Illinois boar studs for domestic and international sale of frozen sperm. Evaluation with both manual microscopy and CASA methods were conducted to verify quality. A preliminary breeding trial was then conducted to test the fertility of sperm frozen with this method. There were 41 ejaculates from 23 boars used for freezing. Sperm were frozen at 1.4x109 sperm/mL, averaging 55.61.1% (meanSE) motility, following thaw. The samples assessed were not different (P>0.05) in motility when compared with manual or CASA systems, and results were most reliable at a 1:40 sperm dilution. In the preliminary breeding trial, gilts (n=14) were inseminated with either a single (n=10) or double (n=4) AI using 1, 2, or 4x109 motile, frozen-thawed sperm. Overall, the resulting pregnancy rates averaged 71.4% and numbers of normal fetuses per litter averaged 15.51.3 per litter. A feasibility study for freezing cost per ejaculate was estimated at $275/ejaculate or $11/dose of frozen-thawed semen at standard doses of 5x109 total frozen-thawed sperm. This cost estimate did not include genetic value, fixed equipment costs, depreciation, or variable lab space fees. Objective 2 focused on the proper methods for breeding with frozen-thawed boar sperm to achieve fertility. Our hypothesis was that increased numbers of inseminations and increased numbers of motile frozen-thawed sperm would improve pregnancy rate and litter size. Results showed acceptable fertility at high sperm numbers, but also the optimal method for insemination with the lowest dose tested. Gilts (n=111) responded to synchronization methods and were bred with 1, 2, or 4x109 motile frozen-thawed sperm from six boars using a single AI at 32 h, or a double AI, with the first AI at 24 and 32 h following estrus. Ultrasound was conducted at 12 h intervals to estimate the time of ovulation. On day 32 of gestation, overall pregnancy rate (73%) and number of normal fetuses per litter (10.80.5) across all treatments did not differ, and were not affected by number of motile sperm, or the interaction of number of motile sperm and number of inseminations. However, the number of inseminations tended to affect (P=0.14) the number of normal fetuses. Litter size increased with a double AI compared to single AI. Multiple inseminations helped to allow insemination to occur close to ovulation in response to variation in the time of ovulation. Both pregnancy rate and number of normal fetuses were greater when the time of the AI at 32 h occurred closer to the estimated time of ovulation (P<0.05). In addition, other factors such as presence of an abnormal ovary at day 30 decreased (P<0.001) pregnancy rate, while boar affected number of normal fetuses (P<0.01). Analysis of our data using a fertility index revealed doses of 2x109 motile sperm with multiple AI can achieve acceptable fertility with use of less sperm, when compared to AI using 4x109 motile sperm. The methods described here will investigate the potential for improved fertility when using frozen-thawed sperm, while accounting for variation in time of ovulation.

Adição de Ácido Linoleico conjugado (CLA) no diluidor de congelação de sémen de touro, recolhido Post-Mortem, das raças Holstein Frísia, Aberdeen Angus, Charolês e Limousine

Dissertação de Mestrado, Engenharia Zootécnica, 7 de Abril de 2016, Universidade dos Açores.

Efecto de la jerarquía social sobre la calidad y la congelabilidad del semen de carnero

Las majadas de ovinos funcionan socialmente dentro de un orden jerárquico que se encuentra formado por individuos dominantes (alfa) y subordinados (omega). El macho dominante es el encargado de la reproducción (Ungerfeld & Núñez, 2011). El animal más agresivo es el que ocupa la posición más alta y no está sujeto a las mismas presiones experimentadas por el resto de los miembros. La sexualidad y el comportamiento agresivo cambian en relación al ciclo reproductivo y al fotoperiodo. Así, los animales menos afectados por el fotoperiodo permanecen más agresivos y con mayor incremento en la libido, mientras que la actividad sexual de los animales de baja jerarquía es generalmente suprimida en presencia de los dominantes (Aguirre, 2004). Cuando se insemina artificialmente se busca calidad y congelabilidad seminal que será proporcionada por un buen carnero independientemente de su ranking social en el grupo. (Guerrero et al., 2009). El objetivo de este trabajo fue evaluar el efecto de la jerarquía social sobre la calidad seminal y la congelabilidad del eyaculado de carneros alfa y omega.

Comparación de dos medios para congelar semen de toro y la influencia de tres tiempos de equilibración en la calidad post-descongelación

Los factores que influyen en la crioconservación de semen son la calidad, el manejo y los métodos de congelación (Lozano, 2009). La supervivencia y vitalidad espermática post-descongelación en la crioconservación está relacionada con los protocolos de procesamiento del semen y la dilución (Andrabi, 2009). Existen parámetros que permiten valorar la calidad seminal como la Motilidad Individual Progresiva (MIP), la Vitalidad Espermática (VE) y la tasa de Anormalidades en el semen descongelado que se ven afectados por los tiempos de equilibrio. Además la motilidad espermática y la vitalidad post-descongelación se pueden ver afectadas con el uso de los diferentes diluyentes tanto sintéticos (lecitina de soya) como orgánicos (yema de huevo). El objetivo fue comparar la eficacia del diluyente AndroMed® (TRIS + Lecitina de soya) y Triladyl® (TRIS + Yema de Huevo) y tres tiempos de equilibramiento de acuerdo a los protocolos establecidos en la crioconservación de semen de toro.

Effects of Ageratum Conyzoides on Semen Characteristics and Sperm Morphology in Rats Exposed to Sodium Arsenite

Arsenite is a major environmental toxicant that is well known to cause reproductive injury. The sperm protective potential of Ageratum conyzoides Linn in arsenic-treated rats was carried out in this study taking advantage of the antioxidant constituents and its androgenic activities. Twenty-four male albino rats aged 16 weeks, weighing 225 to 228g were used. They were grouped into 4(A-Da) with each group containing 6 rats. Group A was orally treated with 100mg/kg ethanol leaf extract of Ageratum conyzoides L., daily for 14 days, group B (single oral dose of sodium arsenite 2.5 mg/kg body weight), C (Ageratum conyzoides extract daily for 14 days and sodium arsenite (SA) given on the 14th day) and group D (Propylene glycol as negative control). It was observed that group B had a more lower (p<0.05) percentage motility (26.7±6.67%) when compared across the groups while group A had a significantly higher (p<0.05) mean value (63.3±3.33%). The sperm motility of rats in group D was significantly higher (p<0.05) than groups B and C. This implies that A. conyzoides extract had no adverse effect on the sperm motility of the rats and also ameliorates the adverse effect of arsenite on sperm motility. The mean value obtained for sperm liveability, semen volume and Sperm concentration followed a similar pattern although, the differences were not significant (p>0.05) for semen volume and the Sperm concentration of rats across the groups. The total sperm abnormality obtained across the groups ranges between 10.44 and 14.27% with group B treated with sodium arsenite (SA) having the highest value when compared with groups A and D, although, the differences were not significant (P>0.05). The study concluded that ethanol leaf extract of Ageratum conyzoides has no negative effect on sperm motility, liveability characteristics and morphology and also protected spermatozoa against arsenic reproductive toxicity in wistar strain albino rats..

Comparación del efecto de la yema de huevo de pata y de gallina sobre parámetros espermáticos en semen canino refrigerado

Tesis (Médico Veterinario). -- Universidad de La Salle. Facultad de Ciencias Agropecuarias. Programa de Medicina Veterinaria, 2014

Semen profiles of young men involved as bicycle taxi cyclists in Mangochi District, Malawi: A case-control study

Aim The purpose of this study was to evaluate the semen profiles of bicycle taxi cyclists and healthy controls in Mangochi district, Malawi. Methods Semen samples were collected from young bicycle taxi cyclists after two to three days of sexual abstinence. A control group, comprising young men who were not bicycle taxi operators also submitted semen samples. Samples were left to liquefy for 30 minutes before measurements were conducted of volume, concentration, total motility, and progressive motility. This was followed by preparation of morphology slides. Light microscopy was used for sperm analysis. Results Semen parameters such as volume (1.66 ± 0.18 mL vs. 3.64 ± 0.17 mL; p = 0.0001), concentration (28.31 ± 4.33 x 106/mL vs. 54.95 ± 5.93 x 106/ mL; p = 0.02) , total motility (56.98% ± 8.22% vs. 56.98% ± 8.22%; p = 0.03), progressive motility (22.57% ± 3.35% vs. 59.69% ± 4.82%; p = 0.004), and morphology (6.98% ± 3.23% vs. 19.73% ± 2.32%; p = 0.006) were significantly reduced in the bicycle taxi cyclists compared to the healthy controls. Conclusion In this case-control study, bicycle taxi operators had lower semen volume, concentration, total motility, and progressive motility, as well as a higher concentration of abnormally shaped spermatocytes, compared to healthy controls.

Eficiencia comparativa entre dos diluyentes para la criopreservacion de semen en toros brahmán en el departamento de Antioquia

Tesis (Médico Veterinario). -- Universidad de La Salle. Facultad de Ciencias Agropecuarias. Programa de Medicina Veterinaria, 2014

Evaluar el efecto crioprotector de la dimetilformamida en diferentes tiempos de adición en la criopreservación de semen en caballos criollos colombianos

Bogotá (Colombia): Universidad de La Salle. Facultad de Ciencias Agropecuarias. Maestría en Ciencias Veterinarias

Ajuste del tiempo de inseminación con semen sexado en vacas superovuladas

En la actualidad los resultados de la combinación del semen sexado y superovulación (SOV) no han sido muy alentadores debido a la baja cantidad de espermatozoides así como a la corta vida media de los mismos. En un trabajo previo realizado en Brasil en vacas Holstein, se demostró que usando el protocolo de SOV P36/Lh60 e inseminando con semen sexado a las 18 y 30 horas de aplicado el inductor de la ovulación (intervalo de 12 hs entre inseminaciones) se obtiene igual cantidad de estructuras transferibles que inseminando a las 12 y 24 horas con semen no sexado. El objetivo de esta investigación fue ajustar las horas de la inseminación artificial (IA) en vacas superovuladas e inseminadas con semen sexado para lograr mejor sincronía con las ovulaciones y así aumentar la cantidad de embriones transferibles. Este ajuste evaluó 2 momentos de IA en los cuales el intervalo entre ambas se disminuyó a 6 hs. Para esto se usaron 30 vacas Holstein en producción superovuladas con el protocolo P36/LH60 y se dividieron en tres grupos en forma aleatoria; en el grupo IA18/30 (control; n=10) las inseminaciones se realizaron a las 18 horas y 30 hs de la aplicación del inductor de la ovulación (GnRh); en el grupo IA18/24 (n=10) sé inseminó a las 18 y 24 hs de la GnRH y al grupo IA24/30 (n=10) se inseminó a las 24 y 30 (se usó 2.1x106 espermatozoides sexados / inseminación). No se encontró diferencia estadística en los tres grupos, sin embargo el grupo IA18/24 mostró ventaja numérica sobre el grupo IA24/30 y el Control (4,1±1,5 vs 1,3±0,4 y 1,9±0,6 respectivamente) en la cantidad de embriones transferibles, con lo que concluimos que el ajuste en las horas de IA con semen sexado en vacas Holstein superovuladas puede ser usado con resultados similares a los trabajos anteriores

Comparación de técnicas de colecta seminal: colecta de semen de epidídimo, colecta tradicional y criopreservación, en relación a la calidad espermática, en caninos

Bogotá (Colombia): Universidad de La Salle. Facultad de Ciencias Agropecuarias. Programa de Medicina Veterinaria

Beneficios y ventajas de la inseminación artificial utilizando semen congelado en programas de reproducción en equinos

Bogotá (Colombia): Universidad de La Salle. Facultad de Ciencias Agropecuarias. Programa de Medicina Veterinaria

Artificial fertilization of oocytes and sperm activation in pacu: effects of the spermatozoa:oocyte ratio, water volume, and in natura semen preservation

Objetivou-se foi avaliar a fertilização artificial e a duração da motilidade espermática em pacus com diferentes doses inseminantes, volumes de água e preservação do sêmen in natura. Foram realizados quatro experimentos para avaliação do efeito de doses inseminantes (7x10³, 7x10(4), 7x10(5), 7x10(6) e 7x10(7) espermatozoides ovócito-1) sobre a fertilização artificial dos ovócitos; do efeito do volume de água (0,5; 15,0; 30,0; 45,0 e 60,0 mL de água mL-1 de ovócitos) com doses inseminantes de 105.481 e 210.963 espermatozoides ovócito-1; do efeito de diluição do sêmen (0,005; 0,05; 0,5 e 5,0 µL de sêmen mL-1 de água) sobre a duração da motilidade espermática; e do efeito do armazenamento a 15 ºC por 9 h sobre a duração da motilidade espermática e o índice de sobrevivência espermática. Os maiores resultados obtidos foram: doses inseminantes entre 7x10³ e 7x10(7) espermatozoides ovócito-1; 15 a 60 mL de água mL-1 de ovócitos; diluição de 0.005 µL sêmen mL-1 de água e 98,65% de sobrevivência espermática até o tempo de preservação de 2h45min36s. A preservação a 15ºC por 9 horas não influencia a duração da motilidade espermática. As maiores taxas de fertilização podem ser observadas no emprego de 0,27 a 270 µL de sêmen mL-1 de ovócitos, com 15 a 60 mL de água para ativação.

Desenvolvimento de embriões ovinos produzidos in vitro com semen fresco e congelado usando o método de seleção espermática mini percoll.

2015

Ractopamine stability in meat and bone meal samples store under freezing and ambient conditions.

2016