Colloidal Centrifugation for Dog Sperm Selection Prior Cryopreservation with Glycerol.

Background: Dog ejaculates are produced in three fractions: the first and third fractions contain prostatic fluid, which is harmful for cryopreservation. The second fraction is spermatozoa-rich. Collecting the second fraction of the ejaculate is complex; hence, colloidal centrifugation plus the addition of glycerol at three different concentrations (4, 6, and 8%) as cryoprotectant was evaluated as a method for sperm selection. Methods: In this study, 72 seminal samples from 6 crossbred dogs were evaluated at the University of Cuenca-Ecuador. Two aliquots were made from each sample, one for conventional centrifugation (Group 1), the other for colloidal centrifugation (Group 2). After centrifugation, each aliquot was subdivided into three parts, and 4, 6, and 8% glycerol were added, respectively. Three sperm evaluations were made (initial, pre-thawing, and post-thawing). Results: Sperm concentration was similar between the initial sample (IS) and Group 1 (P>0.05) after centrifugation, whereas fewer spermatozoa where observed in Group 2 (P <0.05). Evaluation of pre-thawing individual progressive motility (IPM) revealed that the spermatozoa in Group 1 underwent a 30% motility reduction in comparison to the IS and Group 2. Post-thawing IPM within the groups with different concentrations of glycerol (4, 6, and 8%) which underwent previous centrifugation with Percoll (Group 2), showed statistical differences (P <0.05). However, the HOS test produced similar percentages. Conclusions: Colloidal centrifugation prior freezing ensures better sperm selection and purification.Background: Dog ejaculates are produced in three fractions: the first and third fractions contain prostatic fluid, which is harmful for cryopreservation. The second fraction is spermatozoa-rich. Collecting the second fraction of the ejaculate is complex; hence, colloidal centrifugation plus the addition of glycerol at three different concentrations (4, 6, and 8%) as cryoprotectant was evaluated as a method for sperm selection. Methods: In this study, 72 seminal samples from 6 crossbred dogs were evaluated at the University of Cuenca-Ecuador. Two aliquots were made from each sample, one for conventional centrifugation (Group 1), the other for colloidal centrifugation (Group 2). After centrifugation, each aliquot was subdivided into three parts, and 4, 6, and 8% glycerol were added, respectively. Three sperm evaluations were made (initial, pre-thawing, and post-thawing). Results: Sperm concentration was similar between the initial sample (IS) and Group 1 (P>0.05) after centrifugation, whereas fewer spermatozoa where observed in Group 2 (P <0.05). Evaluation of pre-thawing individual progressive motility (IPM) revealed that the spermatozoa in Group 1 underwent a 30% motility reduction in comparison to the IS and Group 2. Post-thawing IPM within the groups with different concentrations of glycerol (4, 6, and 8%) which underwent previous centrifugation with Percoll (Group 2), showed statistical differences (P <0.05). However, the HOS test produced similar percentages. Conclusions: Colloidal centrifugation prior freezing ensures better sperm selection and purification

Influence of Penile Spicules of Covies(Cavia porcellus)on their Sexual Behavior, Fertility and Sperm Quality

This research took place in canton Cuenca, province of Azuay, Ecuador, located on UTM 717386 x 9675751, 2 714 meters above sea level, with a mean temperature of 14°C. The study determined the influence of penile spicules on sexual behavior, fertility and sperm quality parameters in cavies (Cavia porcellus), raised on the Ecuadoran highlands. A number of 5 whole males (with penile spicules), and other 5 males with their spicules removedwere included in the study, along with 40 nulliparous females, type A, pelage type 1. The pregnancy percentage of females that copulated with extirpated males decreased 65% in relation to the control group (P<0.05). However, the sexual behavior pattern and sperm quality of the two groups in the study were similar (P>0.05). Therefore, the study concluded that extirpation of the penile spicules has effects on fertility, but not on sexual behavior and sperm quality

Centrifugación coloidal como método de selección espermática previo a la crioconservación con glicerol en caninos

Background: The ejaculate of the dog has three fractions, the first and third containing prostatic fluid that is harmful to cryopreservation, and the second rich in sperm. Obtaining only the second fraction of the ejaculate is complicated; therefore, colloidal centrifugation was evaluated as a sperm selection method, plus the addition of glycerol in three concentrations 4, 6 and 8 % as cryoprotector.  Methods: The study was conducted at the University of Cuenca-Ecuador, 72 seminal samples from 6 crossbreed dogs were evaluated. Each sample was divided into 2 aliquots, the first for Group 1 (conventional centrifugation) and the other Group 2 (colloidal centrifugation), once centrifuged each aliquot was sub-divided into three and 4, 6 and 8 % of glycerol were added. Three spermatic evaluations were carried out (initial, pre and post thawing).  Results: The sperm concentration after centrifugation was similar between the initial sample (MI) and Group 1 (P > 0.05); in Group 2 less sperm was observed (P < 0.05). When assessing the progressive individual motility (MIP) pre freezing, it was observed that the sperm of the Group 1 reduced 30 % its motility in relation to the sample MI and that of Group 2. Post thawing MIP within the groups with different concentrations of glycerol (4, 6, 8 %), which previously received centrifugation with Percoll (Group 2) presented statistical difference (P < 0.05). However, the HOS test gave similar percentages.  Conclusions: It is concluded that colloidal centrifugation, prior to the freezing process, is a technique that allows greater selection and sperm purification.Antecedentes: El eyaculado del perro presenta tres fracciones, la primera y tercera que contienen líquido prostático que es perjudicial para la criopreservación, y la segunda rica en espermatozoides. Obtener únicamente la segunda fracción del eyaculado es complicado; por lo tanto, se evaluó la centrifugación coloidal como método de selección espermática, más la adición de glicerol en tres concentraciones 4; 6 y 8 % como crio protector.  Métodos: El estudio fue realizado en la Universidad de Cuenca-Ecuador, se valoraron 72 muestras seminales procedentes de 6 perros mestizos. Cada muestra fue dividida en 2 alícuotas, la primera para el Grupo 1 (centrifugación convencional) y la otra Grupo 2 (centrifugación coloidal). Una vez centrifugadas cada alícuota fue sub-dividida en tres y se les adicionó 4; 6 y 8 % de glicerol. Se realizaron 3 evaluaciones espermáticas (inicial, pre y post descongelación).  Resultados: La concentración espermática luego de la centrifugación fue similar entre la muestra inicial (MI) y Grupo 1 (P > 0,05); en Grupo 2 se observó menos espermatozoides (P < 0,05). Al valorar la motilidad individual progresiva (MIP) pre congelación, se observó que los espermatozoides del Grupo 1 redujeron un 30 % su motilidad en relación a la muestra MI y la de Grupo 2. La MIP post descongelación dentro de los grupos con diferentes concentraciones de glicerol (4; 6 y 8 %), que recibieron previamente centrifugación con Percoll (Grupo 2) presentaron diferencia estadística (P < 0,05). Sin embargo, la prueba de HOS proporcionó porcentajes similares.  Conclusiones: Se concluye que la centrifugación coloidal, previa al proceso de congelación es una técnica que permite mayor selección y purificación espermática

Espículas peneanas del cobayo (Cavia porcellus), influencia sobre el comportamiento sexual, fertilidad y calidad espermática

La investigación se realizó en el cantón Cuenca, provincia del Azuay-Ecuador. Coordenadas UTM 717 386 x 967 5751, altura 2 714 m s.n.m., temperatura promedio de 14 °C . El estudio determinó la influencia de las espículas peneanas sobre el comportamiento sexual, la fertilidad y parámetros de calidad espermática en cobayos (Cavia porcellus) criados en el altiplano ecuatoriano. Se utilizaron 5 machos enteros (con espículas peneanas), 5 machos extirpados las espículas y 40 hembras nulíparas, conformación Tipo A y pelaje tipo 1. Se determinó que el por ciento de preñez de las hembras copuladas con machos extirpados las espículas peneanas disminuía en 65 % en relación al grupo control (P < 0,05). Sin embargo, el patrón de comportamiento sexual y la calidad espermática de los dos grupos en estudio fue similar (P > 0,05). Por lo tanto, se concluye que la extirpación de las espículas peneanas influye en la fertilidad, pero no en el comportamiento sexual y calidad espermática.Abstract This research took place in canton Cuenca, province of Azuay, Ecuador, located on UTM 717 386 x 967 5751, 2 714 meters above sea level, with a mean temperature of 14 °C . This study determined the influence of penis spicules on sexual behavior, fertility and sperm quality parameters in cavy (Cavia porcellus), raised on the Ecuadorian highlands. A number of 5 whole males (with penis spicules) were included in the study. Also, included were 5 males with their spicules removed, and 40 multiparous females, conformation type A, pelage type 1. The pregnancy percentage of females that copulated with extirpated males decreased 65 % in relation to the control group (P < 0.05). However, the sexual behavior pattern and sperm quality of the two groups in the study were similar (P > 0.05). Therefore, the study concluded that extirpation of the penis spicule has effects on fertility, but not on sexual behavior and sperm quality

Control de la emergencia de la onda folicular mediante la aplicación de benzoato de estradiol, GnRH y ablación folicular en el metaestro tardío en vaquillas de trópico alto

The response of the dominant follicle (DF), the profile of the FSH and estradiol and the onset of the follicular wave after the application of estradiol benzoate, GnRH, or follicular ablation in late metestrus were characterized in crossbred Holstein heifers, reared at 2500 meters above sea level and fed to grazing. Nine animals weighing 427.7±23.0 kg, body condition 2.7±0.1, age 26.7±1.4 months were used. Before the experiment, the heifers were synchronized and day 6 of the oestrous cycle was considered day zero (start of the experiment). There were three treatments: T1 = Oestradiol benzoate - BE (n=9), T2 = GnRH (n=9), T3 = Ablation (n=9). The evaluation of the DF was carried out by ultrasound every 24 hours, for 5 consecutive days. FSH and oestradiol levels were analysed every 5 days from day 0 by radioimmunoassay. The onset of the follicular wave and number of follicles recruited were determined retrospectively when assessing the daily ultrasound. The application of BE (T1) caused an initial growth of the DF, then it was maintained and finally went to atresia, the peak of FSH appeared at 48h (0.87±0.21 ng/ml) and the onset of the follicular wave was at 3.2±0.15 d post-treatment. In T2, 55.5% of the heifers ovulated in 24h and 44.5% between 24-48h, the peak of FSH was at 24h post-treatment and the onset of the wave in 2.4±0.18 d. The T3 heifers showed the FSH peak at 24h and wave restarted at 1.8±0.15 days. It is concluded that the application of BE in the heifers of the study favoured then an earlier restart of the follicular wave. Besides, the presence of corpus luteum and DF prior to the application of GnRH and ablation allows a more solid and homogeneous restart of the follicular wave.Se caracterizó el comportamiento del folículo dominante (FD), el perfil de la FSH y estradiol, el momento del reinicio de la onda folicular pos-aplicación de benzoato de estradiol, GnRH, o ablación folicular en el metaestro tardío en vaquillas Holstein mestizas, sobre los 2500 msnm y alimentadas al pastoreo. Se utilizaron nueve animales de peso 427.7±23.0 kg, condición corporal 2.7±0.1, edad 26.7±1.4 meses. Previo al experimento las vaquillas fueron sincronizadas y se consideró día cero (inicio del experimento) al día 6 del ciclo estral. Se tuvo tres tratamientos: T1=Benzoato de estradiol - BE (n=9), T2=GnRH (n=9), T3=Ablación (n=9). La evaluación del FD se realizó mediante ecografía cada 24h, durante 5 días consecutivos. Los niveles de FSH y estradiol fueron analizados cada 5 días a partir del día 0 mediante radioinmunoensayo. El reinicio de la onda y número de folículos reclutados fueron determinados en forma retrospectiva al valorar los resultados de las ecografías diarias. La aplicación de BE (T1) provocó un crecimiento inicial del FD, luego se mantuvo y finalmente fue a la atresia, el pico de FSH apareció a las 48h (0.87±0.21 ng/ml), el reinicio de la onda folicular fue a los 3.2±0.15 d pos-tratamiento. En T2, el 55.5% de las vaquillas ovularon en 24h y el 44.5% entre las 24-48h, el pico de FSH fue a las 24h pos-tratamiento y el reinicio de la onda en 2.4±0.18 d. Las vaquillas de T3 presentaron el pico de FSH a las 24h y reinicio de onda 1.8±0.15 días. Se concluye que la aplicación de BE en las vaquillas del estudio causa que reinicien tempranamente la onda folicular. Además, la presencia de cuerpo lúteo y FD previo a la aplicación de GnRH y ablación permite un reinicio de la onda folicular más sólido y homogéneo

Tamaño del folículo ovulatorio, cuerpo lúteo y progesterona sanguínea en vaquillas receptoras de embriones de tres razas en pastoreo en Ecuador.

El objetivo del trabajo fue determinar el tamaño del folículo preovulatorio del cuerpo lúteo y los niveles de progesterona sanguínea en los días 6 y 12 post-ovulación en vaquillas de las razas Holstein, Brown Swiss  y Criolla  criadas en pastoreo en el Altiplano Ecuatoriano. Se determinó el tamaño del folículo preovulatorio, del cuerpo lúteo y los niveles sanguíneos de progesterona en los días 6 y 12 post-ovulación en nueve vaquillas de cada raza. La valoración de las estructuras ováricas se realizó mediante ultrasonografía. La concentración de progesterona sanguínea se determi-nó con el método de electroquimio-luminiscencia. Se estableció que el tamaño del folículo preovulatorio de las vaquillas Criolla  fue más grande (14,6  ±  0,41 mm) que el de las Holstein  (12,7  ±  0,47 mm)  y Brown Swiss  (12,7  ± 0,65 mm). La concentración de progesterona en los días 6 y 12  post-ovulación  en las vaquillas Criolla  (11,0  ±1,68 ng/ml y 18,4 ± 2,04 ng/ml, respectivamente) fue más elevada que el de las Holstein y Brown Swiss. Se observa-ron valores de  asociación lineal altos y significativos (P  <  0,05), entre  el  folículo preovulatorio;  cuerpo lúteo  y progesterona en las  Holstein  y Brown Swiss  y se demostró que  por cada milímetro de tamaño adicional sobre la media del Criolla  del  día  6, se generaría 1,67 ng/ml más de progesterona en las vaquillas Criolla  (P  <  0,05) que en las dos restantes. Se concluye que las vaquillas Criolla presentaron mayores niveles de progesterona en los días 6 y 12 post-ovulación en comparación con las razas Holstein y Brown Swiss, lo que generaría un mejor ambiente uterino y, por consiguiente, menores pérdidas embrionarias tempranas.Size of Ovulation Follicle and Corpus Luteum, and Blood Progesterone in Heifers Receiving Em-bryos from Three Cattle Breeds Grazing, in Ecuador. ABSTRACTThe aim of this paper was to determine the size of the pre-ovulation  follicle in the  corpus luteum, and in  bloodprogesterone on the sixth and twelfth days  after ovulation, in Holstein, Brown, Swiss and Criolla breeds grazing on Ecuadoran Highlands. The size of the pre ovulation  follicle  and  corpus luteum, and the levels of blood progesterone were determined on days sixth and twelfth, in  nine heifers from each breed.  Assessment of ovarian structures was made by ultrasound scanning. The concentration of blood progesterone was determined by  electrochemilumines-cence. The pre ovulation  follicle  in Criolla heifers was larger (14.6 ± 0.41 mm)  than  in  Holstein (12.7 ±0.47 mm), and in Brown Swiss (12.7 ± 0.65 mm).  Progesterone concentration on days six and twelve  after  ovulation of Criolla heifers (11.0 ± 1.68 ng/ml,  and 18.4 ± 2.04 ng/ml, respectively) was higher than  for Holstein and Brown Swiss.  The linear association values were high and significant (P < 0.05), in the pre ovulati on follicle, corpus luteum and proges-terone, in Holstein and Brown Swiss. It was demonstrated that for every additional millimeter above the Criolla mean on the sixth day, 1.67 ng/ml more of progesterone was generated in Criolla heifers (P < 0.05) than in the rest. It was concluded that the Criolla heifers had higher progesterone levels  than  Holstein and Brown Swiss  on the sixth and twelfth days, thus producing better conditions in the uterus and lower early embryo losses

Effect of extirpation of guinea pig (Cavia porcellus) penile spicules on carcass yield compared to chemical castration with 2% iodized alcoho

The aim of this study was to determine and compare the effect of the removal of the spicules of the penis of guinea pig and chemical castration on weight after 12-hour fast pre slaughter and carcass yield. 72 guinea pigs Type I between 35- 45 days of age were used, the same that were distributed in three treatments, T1 whole animals without castration, T2 animals removed spicules glans and T3 chemically castrated animals with iodized alcohol 2% intratesticular; removal and castration was performed in the fifth week of age. Live weight loss by fasting 12 hours pre slaughter, carcass yield relative to live weight at the end of the production process and post-fasting through design covariance initial weight was evaluated. The weight lost by fasting for 12 hours before slaughter showed no REDVET Rev. Electrón. vet. http://www.veterinaria.org/revistas/redvet 2018 Volumen 19 Nº 4 - ¿http://www.veterinaria.org/revistas/redvet/n040418.html Efecto de la extirpación de las espículas del pene de cuy (Cavia porcellus) sobre el rendimiento a la canal en comparación con la castración química con alcohol yodado al 2% http://www.veterinaria.org/revistas/redvet/n040418/041805.pdf 2 significant difference (P≥0.05) between treatments being lower for T2 (108.62 g.); the carcass performance relative to body weight before and after fasting shows significant differences (P<0.05) in favor of T2 (70.82% and 76.23%). This is concluded that the removal of the spicules of the penis in guinea pigs improves performance at the carcas

Accuracy of ultrasound and visual inspection in antral follicular count in crossbred Holstein cows raised under grazing systems at high altitude

4 Pág.The objective of this research was to investigate the diagnostic accuracy of post-mortem ultrasound in antral follicle count (AFC) determination and compare it with visual AFC in grazing crossbred Holstein cows, at high altitude in Ecuador. Pre-mortem blood from 80 cows was collected, and AFC and ovarian characteristics were analysed post-mortem by ultrasound and visual techniques. AFC counts were stratified as high, medium or low by terciles. Mean AMH concentration in pre-mortem blood was 280.1 ± 15.53 pg/mL. The AFC obtained by visual inspection (26.9 ± 9.49 follicles) was 23.8% higher than by ultrasound (20.5 ± 7.53 follicles) in all ovaries. Body condition score, age and weight of the cattle did not interact with the count technique. In the low AFC group, visual inspection and ultrasound provided similar AFC results. However, in the Medium- and High-AFC groups, AFC by ultrasound was 14.9% lower than AFC by visual inspection. We confirm that ultrasound can be used with great accuracy for AFC >3 mm (close to the resolution limit) in grazing crossbred Holstein cows at high altitude.Work financed by Research Directorate University Cuenca, Ecuador (Project-No. UC_XVII_2018_37_Ayala_Luis).Peer reviewe

Colloidal Centrifugation as a Method of Sperm Selection Prior to Cryopreserva-tion with Glycerol in Canines

Antecedentes: El eyaculado del perro presenta tres fracciones, la primera y tercera que contienen líquido prostático que es perjudicial para la criopreservación, y la segunda rica en espermatozoides. Obtener úni-camente la segunda fracción del eyaculado es complicado; por lo tanto, se evaluó la centrifugación coloi-dal como método de selección espermática, más la adición de glicerol en tres concentraciones 4; 6 y 8 % como crio protector. Métodos: El estudio fue realizado en la Universidad de Cuenca-Ecuador, se valoraron 72 muestras semi-nales procedentes de 6 perros mestizos. Cada muestra fue dividida en 2 alícuotas, la primera para el Gru-po 1 (centrifugación convencional) y la otra Grupo 2 (centrifugación coloidal). Una vez centrifugadas ca-da alícuota fue sub-dividida en tres y se les adicionó 4; 6 y 8 % de glicerol. Se realizaron 3 evaluaciones espermáticas (inicial, pre y post descongelación). Resultados: La concentración espermática luego de la centrifugación fue similar entre la muestra inicial (MI) y Grupo 1 (P > 0,05); en Grupo 2 se observó menos espermatozoides (P < 0,05). Al valorar la moti-lidad individual progresiva (MIP) pre congelación, se observó que los espermatozoides del Grupo 1 redu-jeron un 30 % su motilidad en relación a la muestra MI y la de Grupo 2. La MIP post descongelación den- tro de los grupos con diferentes concentraciones de glicerol (4; 6 y 8 %), que recibieron previamente cen-trifugación con Percoll (Grupo 2) presentaron diferencia estadística (P < 0,05). Sin embargo, la prueba de HOS proporcionó porcentajes similares. Conclusiones: Se concluye que la centrifugación coloidal, previa al proceso de congelación es una técnica que permite mayor selección y purificación espermáticaBackground: The ejaculate of the dog has three fractions, the first and third containing prostatic fluid that is harmful to cryopreservation, and the second rich in sperm. Obtaining only the second fraction of the ejaculate is complicated; therefore, colloidal centrifugation was evaluated as a sperm selection method, plus the addition of glycerol in three concentrations 4, 6 and 8 % as cryoprotector. Methods: The study was conducted at the University of Cuenca-Ecuador, 72 seminal samples from 6 crossbreed dogs were evaluated. Each sample was divided into 2 aliquots, the first for Group 1 (conven-tional centrifugation) and the other Group 2 (colloidal centrifugation), once centrifuged each aliquot was sub-divided into three and 4, 6 and 8 % of glycerol were added. Three spermatic evaluations were carried out (initial, pre and post thawing). Results: The sperm concentration after centrifugation was similar between the initial sample (MI) and Group 1 (P > 0.05); in Group 2 less sperm was observed (P < 0.05). When assessing the progressive indi-vidual motility (MIP) pre freezing, it was observed that the sperm of the Group 1 reduced 30 % its motili-ty in relation to the sample MI and that of Group 2. Post thawing MIP within the groups with different concentrations of glycerol (4, 6, 8 %), which previously received centrifugation with Percoll (Group 2) presented statistical difference (P < 0.05). However, the HOS test gave similar percentages. Conclusions: It is concluded that colloidal centrifugation, prior to the freezing process, is a technique that allows greater selection and sperm purificatio

Espículas peneanas del cobayo (Cavia porcellus), influencia sobre el comportamiento sexual, fertilidad y calidad espermática

La investigación se realizó en el cantón Cuenca, provincia del Azuay-Ecuador. Coordenadas UTM 717 386 x 967 5751, altura 2 714 m s.n.m., temperatura promedio de 14 °C . El estudio determinó la influencia de las espículas peneanas sobre el comportamiento sexual, la fertilidad y parámetros de calidad espermática en cobayos (Cavia porcellus) criados en el altiplano ecuatoriano. Se utilizaron 5 machos enteros (con espículas peneanas), 5 ma-chos extirpados las espículas y 40 hembras nulíparas, conformación Tipo A y pelaje tipo 1. Se determinó que el por ciento de preñez de las hembras copuladas con machos extirpados las espículas peneanas disminuía en 65 % en rela-ción al grupo control (P < 0,05). Sin embargo, el patrón de comportamiento sexual y la calidad espermática de los dos grupos en estudio fue similar (P > 0,05). Por lo tanto, se concluye que la extirpación de las espículas peneanas influye en la fertilidad, pero no en el comportamiento sexual y calidad espermáticaThis research took place in canton Cuenca, province of Azuay, Ecuador, located on UTM 717 386 x 967 5751, 2 714 meters above sea level, with a mean temperature of 14 °C . This study determined the influence of penis spic-ules on sexual behavior, fertility and sperm quality parameters in cavy (Cavia porcellus), raised on the Ecuadorian highlands. A number of 5 whole males (with penis spicules) were included in the study. Also, included were 5 males with their spicules removed, and 40 multiparous females, conformation type A, pelage type 1. The pregnancy per-centage of females that copulated with extirpated males decreased 65 % in relation to the control group (P < 0.05). However, the sexual behavior pattern and sperm quality of the two groups in the study were similar (P > 0.05). Therefore, the study concluded that extirpation of the penis spicule has effects on fertility, but not on sexual behavior and sperm quality