Studies performed on stallions have characterized detrimental effects on semen quality resulting from thermal stress by testicular insulation, but few have investigated the effects of exercise-induced increases in core body temperature on stallion semen parameters. To our knowledge, this is the first study that correlates subcutaneous scrotal temperature and stallion spermatozoa quality using a subdermal scrotal thermal sensory device. Stallions were assigned to a non-exercised (non-ex; control; n=4) or exercised (ex; n=4) group. A motorized equine exerciser was used to work stallions 30 min/d for 4 d/wk during a 12-wk period from July through October. Temperatures (subcutaneous scrotal, subcutaneous neck, rectal, and ambient) were recorded before exercise, immediately after exercise, and 60 and 120 min post-exercise. Humidity data were obtained later to determine THI. No deleterious effects were observed from implantation of thermal sensory devices. An interaction of treatment and time (P < 0.0001) was evident for rectal and neck temperatures. The relationship between scrotal and rectal temperatures was highest (rs =0.761), and other correlations existed between scrotal, neck, and ambient temperatures, as well. Mean rectal temperature in the ex group increased 1.9°C (P < 0.0001), while there was a slight increase in scrotal temperature of 0.8°C (P > 0.05) from 0 min to 22 min. Although an increase in core body temperature was successfully induced by exercise protocol, scrotal temperatures were not significantly affected, and no treatment effects were found in any of the semen parameters measured (P > 0.05). Therefore, no significant changes in fresh or cooled semen parameters resulted from exercise or increases in core body temperature resulting from exercise protocol used in this study. Results of this study indicate that thermal sensory devices are a safe and effective way to measure subcutaneous scrotal and neck temperatures. Although an increase in core body temperature was successfully induced by exercise protocol, scrotal temperatures were not significantly affected, indicating efficient thermoregulation of the testes by the scrotum. Since the testes experienced no significant thermal insult during the exercise protocol, no significant changes in fresh or cooled semen parameters were evident as a result of exercise or elevated core temperature.

No description available.

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.

El objetivo del presente trabajo fue evaluar el efecto del momento de adición del glicerol durante la curva de enfriamiento sobre la calidad espermática del semen canino. Para esto se utilizaron 15 eyaculados de 4 perros adultos. Se evaluó la motilidad, volumen, concentración espermática e integridad funcional de membrana de cada eyaculado. Cada muestra de semen fue sometida a 3 métodos de adición del glicerol (Tratamiento 1, Tratamiento 2 y Tratamiento 3) los cuales se diferenciaron por la concentración de glicerol presente en el dilutor durante la curva de enfriamiento. En el Tratamiento 1 el total del glicerol se adicionó al inicio de la curva de enfriamiento, en el Tratamiento 2 se adicionó una fracción del glicerol total al inicio de la curva de enfriamiento y la fracción restante al final de la curva, finalmente en el Tratamiento 3 el glicerol se añadió al final de la curva de enfriamiento. Finalizada la curva de enfriamiento el semen se envaso en pajillas de 0.5 mL para proceder al congelamiento en nitrógeno liquido. La motilidad progresiva y la integridad funcional de membrana fueron los parámetros utilizados para evaluar la calidad seminal al descongelamiento. No se encontró diferencia estadística entre ninguno de los 3 métodos de adición del glicerol, a pesar de esto el Tratamiento 1 mantenía ligeramente mejor la motilidad y la integridad funcional de membrana (44% y 46%) en comparación con el Tratamiento 2 (37% y 36%) y el Tratamiento 3 (38% y 37%). Esto nos demuestra que el momento de adición del glicerol no influye de manera diferencial sobre la motilidad progresiva y la integridad funcional de membrana pudiéndose gracias a esto simplificar el proceso de criopreservación al añadir el glicerol al inicio de la curva de enfriamiento pues ya no se manipularía el semen hasta el envasado. / The objetive of this experiment was to evaluate the effect of the moment of addition of the glycerol during the curve of cooling on the spermatic quality of the canine semen. For this there were used 15 ejaculated of 4 adult dogs. There was evaluated the motility, volume, spermatic concentration and functional integrity of membrane of every ejaculated. Every sample of semen was submitted to 3 methods of addition of the glycerol (Treatment 1, Treatment 2 and Treatment 3) which differentiated for the concentration of glicerol present in the dilutor during the curve of cooling. In the Treatment 1 the whole of the glycerol was added to the beginning of the curve of cooling, in the Treatment 2 added to himself a fraction of the entire glycerol to the beginning of the curve of cooling and the remaining fraction at the end of the curve, finally in the Treatment 3 the glycerol was added at the end of the curve of cooling. Finished the curve of cooling the semen was packed in straw hats of 0.5 ml to proceed to the freezing in liquid nitrogen. The progressive motility and the functional integrity of membrane were the parameters used to evaluate the seminal quality to the post-thawing. There was not statistical difference between any of 3 methods of addition of the glicerol, in spite of this the Treatment 1 was keeping lightly better the motility and the functional integrity of membrane (44 % and 46 %) compared to the Treatment 2 (37 % and 36 %) and the Treatment 3 (38 % and 37 %). This demonstrates us that the moment of addition of the glycerol does not influence in a distinguishing way the progressive motility and the functional integrity of membrane being able thanks to this to simplify the process of criopreservación after the glycerol to add to the beginning of the curve of cooling since the semen would not be already manipulated up to the stiff one.

Se emplearon 32 muestras de semen procedentes de cuatro ovinos a fin de ser criopreservadas adicionándoles dos antioxidantes:Tempo (2,2,6,6 tetrametil-1-piperidiniloxil) y Tempol (4-hidroxi 2,2,6,6 tetrametil-1-piperidiniloxil), cada uno en concentración de 0.5, 1.0 y 2.5 mM. La finalidad del trabajo fue evaluar el efecto postdescongelamiento que pudiera tener sobre la motilidad progresiva, la viabilidad e integridad acrosomal, y la capacitación espermática prematura. Para cada concentración de cada antioxidante y para el grupo control, se utilizó un dilutor en base a Tris, realizándose 8 repeticiones, cada una con 4 muestras de semen. Los resultados obtenidos demuestran que la adición de Tempo a una concentración de 0.5 mM mejora significativamente la calidad del semen criopreservado en comparación con el grupo control, incrementado los porcentajes de motilidad progresiva (79 vs. 67 %), viabilidad e integridad acrosomal (70 vs. 58 %) y reduciendo la capacitación espermática prematura (9 vs. 15 %). Sin embargo, la adición de Tempol disminuyó la calidad seminal post descongelamiento en comparación con el control. Consecuentemente, el efecto de las especies reactivas de oxígeno sobre la calidad seminal puede ser reducido por la adición de Tempo 0.5 mM durante el enfriamiento. En conclusión la adición de Tempo 0.5 mM en un dilutor en base a Tris, al finalizar la fase de enfriamiento, podría constituir una estrategia para mejorar la calidad de semen ovino criopreservado y por lo tanto aumentar los porcentajes de fertilidad en la inseminación artificial en ovejas. / Trirty-two semen samples coming from 4 rams were frozed using 2 antioxidants, Tempo (2,2,6,6 tetramethyl-1-piperidinyloxyl) and Tempol (4-hidroxi 2,2,6,6 tetramethyl-1-piperidinyloxyl), each one in concentrations of 0.5, 1.0, and 2.5 mM, with the objective of evaluate their effects on post-thaw motility, viability, acrosomal integrity, and earlier sperm capacitation. It were realized 8 replication for each antioxidant concentrantion and control group, using 4 semen samples in each replication. Semen samples were diluted on a Tris extender. Results show that Tempo 0.5 mM improve frozen semen quality (P is less than 0.05) in comparison with control group, by increasing percentages of progresive motility (79 vs. 67 %), viability and acrosomal integrity (70 vs. 58 %), and decreasing earlier sperm capacitation (9 vs. 15 %). However, when Tempol was used, it was observed a decrease in frozed semen quality in comparison with control group. Consequently, the adverse effect of reactive oxygen species on semen quality could be reduced by the addition of Tempo 0.5 mM during the cooling process. In conclusion, the use of Tempo 0.5 mM on a Tris extender, at the end of the cooling process could be an alternative for improving the quality of frozed ram semen, and in this way, improve the fertility in artificial insemination in ewes.

Neutrophils ordinarily infiltrate the female reproductive tract subsequent to mating or artificial insemination, resulting in reduced fertility. Recently, it was demonstrated that equine neutrophil extracellular traps (NETs) entangled sperm in these DNA-rich structures, interfering with their normal transport through the female reproductive tract. Seminal plasma (SP) or proteinaceous extracts from SP inhibited sperm-neutrophil binding and specifically degraded sperm-activated NETs, without suppressing bactericidal activity of neutrophils. Fertility-associated antigen (FAA), a 31 kDa naturally occurring heparin-binding protein (HBP) produced by the accessory sex glands, has been shown to bind to sperm and potentiate heparin-induced capacitation. FAA shares 87% identity with DNase I-like family members, and contains two internal DNase-I-like peptide motifs. The purpose of this study was to determine if a recombinant form of FAA displayed capacitating effects associated with the native protein and to determine whether rFAA displayed DNase activity similar to SP or SP protein extracts to inhibit sperm-neutrophil binding.

No description available.

No description available.

Thesis (MScMed)--University of Stellenbosch, 2005. / Bibliography. Also available via the Internet.

Thesis (MScMedSci)--University of Stellenbosch, 2007. / Bibliography. Also available via the Internet.