Abstract The objective of this study was to evaluate the effect of three doses of equine chorionic gonadotropin (eCG) for ovarian superstimulation on ovarian response, follicular development and cumulus-oocyte complexes (COCs) collection in llamas. For this purpose, eighteen multiparous non-lactating adult (4–7 yo) female llamas with an average body condition of 2.8 (BCS 1–5) were submitted to a follicular ablation (FA) to induce a new follicular wave emergence. Two days after FA (Day 0), synchronized llamas were randomly allocated to three treatment groups (n = 6/group) and given 500, 750 and 1000 IU of eCG (Novormon®, Syntex, Buenos Aires, Argentina) per animal respectively to induce ovarian superstimulation. Transrectal ultrasonography were performed on Days 2, 4, and 6; and ovum pick up (OPU) was performed on Day 6. Data was evaluated by one-way analysis of variance (ANOVA), repeated measures ANOVA, and 2-tailed Chi-square. The average size (mm) of follicles was greater (p≤ 0.05) in the 1000 IU group compared to the other groups. There was a greater (p≤ 0.05) number of follicles ≥ 7 mm in the 1000 IU group compared to the 500 IU group. Number of COCs collected on Day 6 and the COC recovery rate were not different among groups. In conclusion, a single dose of 1000 IU of eCG induced the best ovarian response resulting in larger and greater number of follicles at the time of OPU. (eCG cumulusoocyte cumulus oocyte (COCs purpose nonlactating non lactating 4–7 47 4 (4– yo 28 2 8 2. BCS 1–5 15 1 5 (FA emergence 0, 0 , 0) n 6/group 6group 75 100 Novormon®, Novormon (Novormon® Syntex Aires Argentina OPU (OPU oneway one way ANOVA (ANOVA) 2tailed tailed Chisquare. Chisquare Chi square. square Chi-square (mm p≤ p (p 0.05 005 05 50 conclusion 4– (4 1– 10 Novormon® (Novormon (ANOVA 0.0 00 ( 0.

Abstract The reproductive efficiency of the herd is correlated with higher productivity in livestock. Reproduction biotechniques, such as ovulation synchronization protocols, are important to optimize production and accelerate genetic profit in beef and dairy herds. The objective of this review is to describe the evolution over the last 40 years of the artificial insemination (AI) and the timed-AI (TAI) protocols in cattle from a Brazilian perspective. TAI protocols are based on synchronizing emergence of the wave of follicular growth, controlling circulating progesterone (P4) concentrations, stimulating the final growth of the follicle and inducing a synchronized ovulation. Hormonal alternatives that optimize the response at the end of the protocol and strategies to induce final follicle growth and ovulation in categories of females with low expression of estrus are described. Furthermore, the potential positive effect of previous exposure to injectable P4 on fertility of Bos indicus and Bos taurus cows is also discussed. livestock biotechniques herds 4 AI (AI timedAI timed (TAI perspective P (P4 concentrations described Furthermore discussed (P

Abstract The aim was to examine the exogenous supply of progesterone, and the influence of the corpus luteum (CL) at the beginning of the estrus synchronization protocols in sheep, for 6 and 11 days, on reproductive variables. The sheep were presynchronized to obtain a premature or functional CL at the beginning of the synchronization period to be evaluated. Estrus presentation was lower in short treatments; This triggered a difference (p < 0.05) in pregnancy rate between short and conventional treatments. On the day of estrus, steroid concentration was different: progesterone was higher in short treatments; while estradiol was lower in the short treatment with CL. The presence of a premature or functional CL at the initiation of the treatments commonly used to synchronize estrus does not modify the expression of the reproductive variables; Nevertheless, in short treatments the expression is affected. It is important to use ultrasonography to corroborate what happened at the ovarian level.

Resumen El objetivo fue evaluar el aporte exógeno de progesterona, y la influencia del cuerpo lúteo (CL) al iniciar protocolos de sincronización de estros en ovejas, por 6 y 11 días, sobre las variables reproductivas. Las ovejas fueron presincronizadas para obtener un CL inmaduro o funcional al iniciar el periodo de sincronización a evaluar. La presentación de estros fue baja en tratamientos cortos; esto desencadenó discrepancias (p < 0.05) en porcentaje de gestación de tratamientos de 6 días y los tradicionales de 11 días. El día del estro la concentración de esteroides fue diferente, progesterona fue mayor en tratamientos cortos; mientras que estradiol fue menor en el tratamiento corto con CL. La presencia de un CL inmaduro o funcional al iniciar tratamientos para sincronizar estro convencionales no modifica la expresión de las variables reproductivas; sin embargo, en tratamientos cortos la expresión se ve afectada. Es importante el uso de ultrasonografía para corroborar lo que pasa a nivel ovárico.

Abstract As our understanding of ovarian function in cattle has improved, our ability to control it has also increased. The development of Fixed-Time Artificial Insemination (FTAI) protocols at the end of the 20th century has increased exponentially the number of animals inseminated over the last 20 years. The main reasons for this growth were the possibility of obtaining acceptable pregnancy rates without heat detection and, above all, the induction of cyclicity in suckled cows in postpartum anestrus and prepubertal heifers at the beginning of the breeding season. Most FTAI treatments in South America have been based on the use of progesterone (P4) releasing devices and estradiol to synchronize both follicular wave emergence and ovulation, with pregnancy rates ranging from 40 to 60%. These protocols are implemented on a regular basis, allowing producers access to high-quality genetics, and increasing the overall pregnancy rates during the breeding season. In addition, it provided the professionals involved in these programs with a new source of income and the diversification of their practices into activities other than their usual clinical work. Many of these practices are now apparently at risk from restrictions on the use of estradiol by the European Union (EU) and other countries. However, the development of alternative protocols based on GnRH, with P4 devices and eCG and other new products that are not in the market yet will allow us to adapt to the new times that are coming. Logically, the challenge has already been raised and we must learn to use alternative protocols to try to continue increasing the use of this technology in beef and dairy herds. The objective of the present review is to describe the main aspects of banning estradiol in livestock production, the negative impacts on reproductive efficiency, and to present some alternative FTAI protocols for dairy and beef cattle. improved FixedTime Fixed Time (FTAI th 2 years all season P (P4 ovulation 4 60 60% basis highquality high quality genetics addition work EU (EU countries However GnRH coming Logically herds production efficiency (P 6

Resumo O hormônio liberador de gonadotrofinas (GnRH) é um decapeptídeo, sintetizado pelos neurônios GnRH no hipotálamo, que estimula a síntese e secreção de gonadotrofinas (hormônio folículo-estimulante, FSH e hormônio luteinizante, LH) da hipófise anterior ou adeno-hipófise. Agonistas de GnRH com estrutura idêntica ou semelhante à natureza estão disponíveis comercialmente desde a década de 1970 e foram inicialmente utilizados para o tratamento de cistos foliculares e melhoria da fertilidade em bovinos (1). A ovulação de folículos ovarianos após a administração de GnRH foi relatada pela primeira vez por Schams et al. (2) e Britt et al. (3), mas foi somente na década de 1990 que o GnRH começou a ser usado para controlar a dinâmica folicular durante o ciclo estral bovino. Sabemos agora que uma nova onda folicular surge dentro de 2 dias após a ovulação do folículo dominante presente no momento do tratamento com GnRH. (4,5) Essa descoberta facilitou o desenvolvimento de protocolos de sincronização baseados em GnRH em combinação com prostaglandinas (PGF) que permitem a inseminação artificial em tempo fixo (IATF), sem a necessidade de detecção de cio. Esses protocolos baseados em GnRH e PGF fazem parte do manejo reprodutivo de bovinos, especialmente bovinos leiteiros, em muitos rebanhos comerciais ao redor do mundo. Existem inúmeros protocolos baseados em GnRH e PGF disponíveis para sincronização de estro e ovulação em bovinos, vários desses protocolos foram explicados e discutidos em detalhes em outros artigos (6,7). O objetivo desta revisão é descrever os fatores associados à liberação de LH e resposta ovulatória após a administração de GnRH, apresentar informações relevantes sobre o efeito dos hormônios esteroides na resposta ao GnRH e, por fim, postular brevemente estratégias que possam melhorar a eficácia de protocolos baseados na aplicação de GnRH e PGF.

Resumen La hormona liberadora de gonadotropina (GnRH) es un decapéptido, sintetizado por las neuronas GnRH en el hipotálamo, que estimula la síntesis y secreción de gonadotrofinas (hormonas folículo estimulante, FSH y luteinizante, LH) de la hipófisis anterior o adenohipofisis. Agonistas de GnRH con una estructura idéntica o similar a la natural han estado disponible comercialmente desde la década del 70 y han sido utilizadas en un principio para el tratamiento de quistes foliculares y el mejoramiento de la fertilidad en el ganado vacuno (1). La ovulación de folículos ováricos después de la administración de GnRH fue reportada por primera vez por Schams et al. (2) y Britt et al. (3), pero no fue hasta la década del 90 cuando la GnRH se comenzó a utilizar para controlar la dinámica folicular durante el ciclo estral del bovino. Ahora sabemos que una nueva onda folicular emerge dentro de los 2 días después de la ovulación del folículo dominante presente en el momento del tratamiento con GnRH (4,5). Este hallazgo facilitó el desarrollo de protocolos de sincronización a base de GnRH en combinación con prostaglandinas (PGF) que permiten la inseminación artificial a tiempo fijo (IATF), sin necesidad de realizar detección de celo. Estos protocolos a base de GnRH y PGF son parte del manejo reproductivo bovino, sobre todo del ganado lechero, en muchos rodeos comerciales del mundo. Existen numerosos protocolos a base de GnRH y PGF disponibles para la sincronización del estro y la ovulación en el ganado, varios de esos protocolos se han explicado y discutido en detalle en otros artículos (6,7). El objetivo de esta revisión es describir los factores asociados a la liberación de LH y respuesta ovulatoria posterior a la administración de GnRH, dar a conocer información relevante sobre el efecto de las hormonas esteroideas en la respuesta a la GnRH y, por último, brevemente postular estrategias que podrían mejorar la eficacia de los protocolos basados en la aplicación de GnRH y PGF.

Abstract The gonadotropin-releasing hormone (GnRH) is a decapeptide, synthesized by the GnRH neurons in the hypothalamus, which stimulates the synthesis and secretion of gonadotrophins (follicle-stimulating hormone, FSH and luteinizing hormone, LH) from the anterior pituitary or adenohypophysis. GnRH agonists with a structure identical or similar to nature have been commercially available since the 1970’s and were initially used for the treatment of follicular cysts and the improvement of fertility in cattle (1). The ovulation of ovarian follicles after GnRH administration was first reported by Schams et al. (2) and Britt et al. (3), but it was not until the 1990’s that GnRH began to be used to control follicular dynamics during the bovine estrous cycle. Studies have shown that a new follicular wave emerges within 2 days after ovulation of the dominant follicle present at the time of GnRH treatment (4,5). This finding facilitated the development of synchronization protocols based on GnRH in combination with prostaglandins (PGF) that allow fixed-time artificial insemination (FTAI), without the need for heat detection. These GnRH-based protocols are part of the reproductive management, especially in dairy cattle, in many commercial herds around the world. There are several GnRH and PGF-based protocols available for estrus and ovulation synchronization in cattle, most of these protocols have been explained and discussed in detail in other publications (6,7). The objective of this review is to describe the factors associated with the release of LH and ovulatory response after the administration of GnRH, to present relevant information on the effect of steroid hormones on the response to GnRH and, finally, to briefly postulate strategies that could improve the efficacy of GnRH and PGF-based protocols.

Resumen Diversos estudios han comprobado que la progesterona tiene un efecto directo en la calidad ovocitaria y el desarrollo inicial embrionario. El objetivo de este estudio fue evaluar el efecto de la concentración de la progesterona (P4) plasmática en el desarrollo de embriones in vitro en vacas de raza Bos taurus taurus (Holstein) y Bos taurus indicus (Gyr). Se utilizaron vacas Holstein y Gyr multíparas, presincronizadas con la aplicación de dos dosis de D-cloprostenol con intervalo de once días. Posteriormente, se distribuyeron en uno de tres grupos experimentales inducidos a diferentes niveles plasmáticos de progesterona. El grupo control fue sincronizado utilizando únicamente D-cloprostenol sin progesterona exógena, el grupo progesterona baja recibió un implante auricular de norgestomet de segundo uso y al grupo progesterona alta se le aplicaron dos implantes auriculares de norgestomet nuevos. Los resultados evidencian que la concentración plasmática de progesterona fue significativamente diferente (P < 0,05) tanto para las vacas Holstein (0,22 ± 0,15, 2,23 ± 0,17 y 5,32 ± 0,22 ng/mL) como para las vacas Gyr (0,24 ± 0,19, 2,05 ± 0,17, 4,85 ± 0,20 ng/mL) entre los grupos control, progesterona baja y progesterona alta, respectivamente. Las vacas Gyr presentaron diferencia (P < 0,05) en la tasa de clivaje del grupo progesterona baja en relación con los grupos control y progesterona alta (87,9 % ± 0,11 vs 77,5 % ± 0,13 y 76,4 % ± 0,10, respectivamente) y en la variable tasa de blastocistos del grupo progesterona baja (48,3 % ± 0,16) en relación con el grupo control (35,8 % ± 0,10) y el grupo progesterona alta (30,4 % ± 0,20). Se concluye que la progesterona tuvo efecto sobre el desarrollo embrionario en vagas de raza Gyr, el cual no se evidenció en las vacas de raza Holstein.

Abstract Studies have shown that progesterone has a direct effect on oocyte quality and early embryonic development. The objective of this study was to evaluate the effect of plasma progesterone (P4) concentration on in vitro embryo development in Bos taurus taurus (Holstein) and Bos taurus indicus (Gyr) cows. Multiparous Holstein and Gyr cows were used, and they were presynchronized with the application of two doses of D-cloprostenol with an interval of 11 days. They were latter distributed in one of three experimental groups induced to different plasma levels of progesterone: control group was synchronized using only D-cloprostenol without exogenous progesterone, low progesterone group received an auricular implant of norgestomet of second use, and high progesterone group received two new auricular implants of norgestomet. The plasma progesterone concentration was significantly different (P < 0.05) for both Holstein cows (0.22 ± 0.15, 2.23 ± 0.17 and 5.32 ± 0.22 ng/mL) and Gyr cows (0.24 ± 0.19, 2.05 ± 0.17, 4.85 ± 0.20 ng/mL) between control, low progesterone, and high progesterone groups respectively. Gyr cows presented differences (P < 0.05) in the cleavage rate of the low progesterone group in relation to the control and high progesterone groups (87.9 % ± 0.11 vs. 77.5 % ± 0.13 and 76.4 % ± 0.10, respectively) and in the variable blastocyst rate of the low progesterone group (48.3 % ± 0.16) in relation to the control group (35.8 % ± 0.10) and the high progesterone group (30.4 % ± 0.20). It is concluded that there is an effect of progesterone on embryonic development of Gyr cows. However, this effect is not evidenced in Holstein cows.

ABSTRACT The aim of this study was to evaluate the superovulatory and embryonic response in alpacas with equine chorionic gonadotropin (eCG) according to the number of follicles recruited >3 mm in the follicular wave (FW). Twenty-six alpacas from the CIP «Chuquibambilla», Universidad Nacional del Altiplano, Puno, Peru, were selected and distributed into two experimental groups: GE01 (n=15 alpacas) with less than three follicles >3 mm and GE02 (n=11 alpacas) with more from four follicles >3 mm in diameter to follicular recruitment post-induction of ovulation by a follicle >7 mm with 0.0042 mg of buserelin acetate (BA). Both groups were evaluated by ultrasound every other day until the beginning of the third follicular wave. Subsequently, females with follicles >7 mm were induced to ovulate with BA (day 0), ovulation was verified at 36 h and 700 IU of eCG was administered. On day 6.5 animals were mated and 7.5 days later the embryos were collected. The superovulatory response was similar in GE01 and GE02 with 4.4 ± 4.0 and 4.7 ± 3.7 follicles between >3 and <7 mm; 7.5 ± 3.7 and 10.1 ± 6.3 follicles >7 mm; and 7.2 ± 3.3 and 8.6 ± 5.0 corpora lutea, respectively. Furthermore, GE01 and GE02 presented 1.6 ± 0.5 and 2.8 ± 1.3 luteinized anovulatory follicles and 2.0 ± 1.0 and 2.0 ± 2.2 anovulatory follicles, respectively. In 80% of alpacas in GE01 2.5 ± 1.4 embryos were collected and in 70% of alpacas in GE02 2.3 ± 1.1 embryos were collected. Likewise, 25 transferable and 5 nontransferable embryos were recovered in GE01 (n=15) and 12 transferable and 4 nontransferable embryos in GE02 (n=10). The results suggest that there is no difference in the ovarian response and embryonic collection in alpacas overstimulated with eCG despite a marked difference in emerging follicles >3 mm in the first FW and that it was not sustainable in the second and third FW.

RESUMEN El propósito del estudio fue evaluar la respuesta superovulatoria y embrionaria en alpacas con gonadotropina coriónica equina (eCG) según el número de folículos reclutados >3 mm en la onda folicular (OF). Se seleccionaron 26 alpacas del CIP «Chuquibambilla», Universidad Nacional del Altiplano, Puno, Perú, para conformar dos grupos experimentales: GE01 (n=15 alpacas) con menos de tres folículos >3 mm y GE02 (n=11 alpacas) con más de cuatro folículos >3 mm de diámetro al reclutamiento folicular pos-inducción de la ovulación de un folículo >7 mm con 0.0042 mg de acetato de buserelina (AB). Ambos grupos fueron evaluados por ecografía cada dos días hasta inicios de la tercera onda folicular. Posteriormente, hembras con folículos >7 mm fueron inducidas a la ovulación con AB (día 0), se verificó la ovulación a las 36 h y se administró 700 UI de eCG. Al día 6.5 se hizo el empadre y 7.5 días después se colectaron los embriones. La respuesta superovulatoria fue similar en GE01 y GE02 con 4.4 ± 4.0 y 4.7 ± 3.7 folículos entre >3 y <7 mm; 7.5 ± 3.7 y 10.1 ± 6.3 folículos >7 mm; y 7.2 ± 3.3 y 8.6 ± 5.0 cuerpos lúteos, respectivamente. Además, GE01 y GE02 presentaron 1.6 ± 0.5 y 2.8 ± 1.3 folículos anovulatorios luteinizados y 2.0 ± 1.0 y 2.0 ± 2.2 folículos anovulatorios, respectivamente. En 80% de alpacas en GE01 se colectaron 2.5 ± 1.4 embriones y en 70% de alpacas de GE02 se colectaron 2.3 ± 1.1 embriones. Así mismo, se colectaron 25 embriones transferibles y 5 no transferibles en GE01 (n=15) y 12 embriones transferibles y 4 no transferibles en GE02 (n=10). Los resultados sugieren que no hay diferencia en la respuesta ovárica y colección embrionaria en alpacas superestimuladas con eCG a pesar de una marcada diferencia de folículos emergentes >3 mm en la primera OF y que no se hizo sostenible en la segunda y tercera OF.

ABSTRACT The ovarian follicular dynamics monitored by transrectal (TR) and transvaginal (TV) routes in successive waves was studied in 10 llamas older than 4 years, multiparous, without calves at foot, body condition of 3.0, during a period of 90 days in the reproductive season. The Sonostar SS8® ultrasound equipment was used with a linear transducer for transrectal examination and endocavitary microconvex transducer for transvaginal examination, both with a frequency of 6.5 MHz. The inter-wave interval was 20 ± 4.4 and 22.5 ± 5.3 via TR and VT, respectively; the number of follicles per cohort was 5.21 ± 0.92 and 6.38 ± 1.26 for TR and TV, respectively (p=0.0011); the maximum diameter of follicles was 12.49 ± 3.57 mm for TR and 13.56 ± 3.91 mm for TV; the growth rate was 0.70 ± 0.35 and 0.67 ± 0.32 mm/day for TR and TV, respectively. The regression equation for the ratio of the maximum follicular diameter and duration of follicular wave for the TR route was y=0.89766x+6.5554 (R²=0.423) and for the TV route y=0.8466x+10.184 (R²=0.479). The duration of the follicular phases was similar between both techniques except for the dominance phase (10.2±3.19 days for TR and 8.67±2.46 for TV, p=0.0418). The relationship between the follicular diameter and the number of follicles detected showed a low negative correlation (R²=-0.0827 for TV and R²=-0.0876 for TR). In conclusion, the transvaginal endocavitary transducer makes it possible to determine with greater precision the characteristics of llama follicular dynamics in their recruitment, dominance and regression phases with overlapping of follicular waves as an important characteristic.

RESUMEN La dinámica folicular ovárica monitoreada por vía transrectal (TR) y transvaginal (TV) en ondas sucesivas en llamas fue estudiada en 10 llamas con más de 4 años, multíparas, sin cría, estado corporal de 3.0, durante un periodo de 90 días en la época reproductiva. Se utilizó el ecógrafo Sonostar SS8® con un transductor lineal para el examen transrectal y transductor microconvexo endocavitario para el examen transvaginal, ambos con frecuencia de 6.5 MHz. El intervalo inter-onda fue de 20±4.4 y 22.5±5.3 por vía TR y TV, respectivamente; el número de folículos por cohorte fue 5.21±0.92 y 6.38±1.26 para TR y TV, respectivamente (p=0.0011); el diámetro máximo de folículos fue 12.49±3.57 mm para TR y 13.56±3.91 mm para TV; la tasa de crecimiento fue de 0.70±0.35 y 0.67±0.32 mm/día para TR y TV, respectivamente. La ecuación de regresión para relación del diámetro máximo folicular y duración de onda folicular para la vía TR fue de y=0.89766x+6.5554 (R²=0.423) y para la vía TV de y=0.8466x+10.184 (R²=0.479). La duración de las fases foliculares fue similar entre ambas técnicas con excepción de la fase de dominancia (10.2±3.19 días para TR y 8.67±2.46 para TV, p=0.0418). La relación entre el diámetro folicular y número de folículos detectados presentó una correlación negativa baja (R²=-0.0827 para TV y R²=-0.0876 para TR). En conclusión, el transductor endocavitario transvaginal permite determinar con mayor precisión las características de dinámica folicular en llamas en sus fases de reclutamiento, dominancia y regresión con sobreposición de ondas foliculares como característica importante.

Abstract The present study aimed to investigate the effects of a combination of progesterone with different doses of E-17β on following end points: (1) ovarian follicular dynamics and emergence of a new follicular wave, and (2) superovulatory response and embryo yield. In Experiment 1, 28 ewes were randomly divided into four groups (n = 7) to receive either 2.0 mg, 1.0 mg, 0.5 mg or none E-17β one day after insertion of a progesterone device. The different doses of estradiol similarly delayed the moment of follicular emergence (overall mean = 3.1 ± 1.0 days vs. control group = 0.86 ± 1.0 days; P < 0.01), but the emergence of the new wave showed greater synchronization with the 0.5 mg dosage of E-17β. In Experiment 2, sixty-two donor ewes received an internal progesterone release device (day -1) for 7 d and 1 d after the insertion of this device (day 0) were allocated randomly to receive 0.5 mg of E-17β or only the vehicle (control group). Superstimulation was initiated on day 3 with the administration of 133 mg of pFSH in eight decreasing doses. Contrary to expectations, the protocol with the administration of 0.5 mg E-17β did not improve the percentage of donors with > 2 CL, the number of CL and the production of embryos (P > 0.05). It was concluded that the combination of progesterone and 0.5 mg E-17β was more efficient in synchronizing the emergence of the new follicular wave, however this approach seems to be unnecessary in ewe’s superovulation programs.

Abstract We aimed to compare the effect of three estradiol benzoate (EB) doses on follicular wave emergence (FWE) and dominant follicle growth of suckled Nelore cows submitted to TAI (D0). On a random day of estrous cycle (D−10), multiparous (MULT; n=36) and primiparous (PRIM; n=20) suckled Nelore cows received an intravaginal progesterone (P4) device and were assigned in three groups. Cows in the EB-1 (n=20), EB-1.5 (n=15) or EB-2 (n=21) groups received, respectively, an im treatment with 1, 1.5 or 2 mg EB. A subgroup (n=10-13 cows/group) were subject to daily ovarian evaluations from D−10 to D0. On D−2, P4 devices were removed, and all cows received the same treatment: 1 mg estradiol cypionate, 0.53 mg sodium cloprostenol, and 300 IU eCG. Statistical analyses were performed considering only the main effects of treatment group and parity order. The proportion of cows with a synchronized FWE and the moment of the FWE did not differ (p>0.05) among the treatment groups (overall: 80% [28/35] and 4.1 ± 0.4 days); however, the FWE occurred earlier (p=0.007) in MULT (3.8 ± 0.2 days) than PRIM (5.1 ± 0.4) cows. The proportion of animals detected in estrus was greater (86% [31/36] vs. 70% [14/20]; p=0.02) and the dominant follicle was larger on D−2 (9.7 ± 0.3 mm vs. 7.8 ± 0.7 mm; p=0.006) and D0 (11.9 ± 0.4 mm vs. 10 ± 0.5 mm; p=0.008) in MULT than PRIM cows. In conclusion, the three EB doses presented similar efficiency to synchronize the FWE in suckled Nelore cows. Moreover, a delayed FWE and smaller dominant follicle is observed in PRIM cows, contributing to the reduced reproductive performance in this parity category when using similar TAI protocols of MULT cows.

The objectives were to investigate the ovarian dynamics and hormonal profiles during the oestrus cycle, and to characterize the ovarian and uterine blood flow (OBF versus UBF) indices on the extent of vascular perfusion towards the dominant follicle (DF) or corpus luteum (CL) during the peri-ovulatory follicular wave in Sahiwal cows. In experiment 1, cyclic cows (n = 21) were selected at their spontaneous oestrus (day 0) and subjected to B-mode ultrasonography throughout the oestrus cycle. In experiment 2, cyclic cows (n = 9) were randomly selected at day 0, and categorized according to the intra-ovarian patterns as ovaries with DF (n = 3), CL (n = 3) or both (DF+CL; n = 3) to characterize the OBF and UBF indices using Doppler ultrasonography. The length of the oestrus cycle (days), number of follicular waves (n) and size (mm) of DF and CL were 20.1 ± 0.9, 2.1 ± 0.2, 14.7 ± 0.7 and 15.9 ± 2.5, respectively. The concentrations (ng/mL) of plasma progesterone increased linearly, and a peak was attained at day 12.2 ± 1.0. However, it reduced linearly with the onset of luteolysis at day 16.4 ± 0.3 of the oestrus cycle. The plasma progesterone (P4) concentrations and the diameter of CL correlated throughout the oestrus cycle. The mean OBF and UBF indices did not differ between intraovarian patterns. Taken together, this information on reproductive physiological parameters could be used to develop synchronization protocols to improve reproductive management in Sahiwal cows.

Resumen El objetivo del actual estudio fue evaluar el efecto del grupo genético de donadoras de ovocitos de las razas Gyr y Holstein sobre el desempeño en la técnica de producción in vitro de embriones. Se utilizaron 12 vacas multíparas, no lactantes, seis de la raza Gyr (Bos indicus) y seis de la raza Holstein (Bos taurus). Previo a la aplicación de la técnica de aspiración folicular, los animales fueron sometidos a sincronización del estro, mediante la administración de 3 mg de benzoato de estradiol (RIC-BE®, Tecnopec Ltda, Brasil), más la inserción de un implante auricular de norgestomet (Crestar®, Intervet, Brasil). La aspiración fue realizada siete días después del inicio del protocolo de sincronización. Los resultados determinaron que las vacas de la raza Gyr presentaron mayor número de folículos visualizados y de ovocitos recuperados, así como mayor tasa de clivaje y de blastocistos en el día siete. Estos resultados permiten considerar que la aplicación de la técnica de producción in vitro de embriones en la raza Gyr puede presentar mejor desempeño en ciertos parámetros en relación con la Holstein.

Abstract The aim of this study was to evaluate the genetic group effect of oocyte donors of the Gyr and Holstein breeds on the performance of the in vitro embryo production technique. Twelve multiparous cows were used, non-lactating, six of the Gyr breed (Bos indicus), and six of the Holstein (Bos taurus) breed. Prior to the implementation of the follicular aspiration technique, the animals were subjected to estrus synchronization through the administration of 3 mg estradiol benzoate (RIC-BE®, Tecnopec Ltda., Brazil) plus the insertion of an ear implant with norgestomet (Crestar®, Intervet, Brazil). Aspiration was performed seven days after the start of the synchronization protocol. The results showed that the cows of the Gyr breed had a higher number of visualized follicles and recovered oocytes, as well as a higher cleavage and blastocyst rate on day 7. These results allow considering that, the application of the in vitro embryo production technique in the Gyr breed offers a better performance in specific parameters in comparison to the Holstein breed.

ABSTRACT 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.

RESUMEN 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.

ABSTRACT The aim of this study was to determine the concentration of glucose, total proteins, albumin, cholesterol and triglycerides in the follicular fluid during the three stages of follicular development in alpacas. Twenty-seven animals from CIP Chuquibambilla, National University of the Altiplano, Puno, Peru were used. The alpaca ovaries were checked by an Aloka SSD 500 ultrasound and a 7.5 MHz linear transducer. The animals were subjected to a follicular wave synchronization procedure in the presence of a follicle >7 mm. On day 9 of the synchronization the animals were distributed according to the stage of the follicular wave in three groups: G1, follicle >7 mm in the growing stage; G2, follicle >7 mm in the static stage; G3, follicle >7 mm in the regression stage. The follicular fluid was aspirated from the dominant follicle with the indicated ultrasound and a 5.0 MHz transvaginal transducer. The fluid was centrifuged, and the supernatant placed in vials and stored in liquid nitrogen. Biochemical determinations were made with specific commercial kits in a semi-automatic biochemical analyser. The results were: glucose 37.04, 41.19 and 30.07 mg/dl; total proteins 1.65, 1.93 and 1.79 g/dl; albumin 1.14, 1.42 and 1.31 g/dl; cholesterol 15.82, 18.3 and 11.08 mg/dl; triglycerides 11.3, 12.35 and 10.78 mg/dl for the stages of growth, static and regression, respectively, without significant differences between stages, with the exception of glucose. The glucose concentration was lower in the regression stage compared to the static stage (p<0.05).

RESUMEN El estudio tuvo como objetivo determinar la concentración de glucosa, proteínas totales, albúmina, colesterol y triglicéridos en el fluido folicular durante los tres estadios del desarrollo folicular en alpacas. Se utilizaron 27 animales del CIP Chuquibambilla, Universidad Nacional del Altiplano, Puno, Perú. Los ovarios de las alpacas fueron revisados mediante un ecógrafo Aloka SSD 500 y un transductor lineal de 7.5 MHz. Los animales fueron sometidos a un procedimiento de sincronización de la onda folicular a la presencia de un folículo >7 mm. En el día 9 de la sincronización fueron distribuidos según el estadio de la onda folicular en tres grupos: G1, folículo >7 mm en estadio de crecimiento; G2, folículo >7 mm en estadio de estática; G3, folículo >7 mm en estadio de regresión. Seguidamente, se procedió a la aspiración del fluido folicular del folículo dominante con el ecógrafo indicado y un transductor transvaginal de 5.0 MHz. El fluido fue centrifugado y el sobrenadante colocado en viales y almacenado en nitrógeno líquido. Las determinaciones bioquímicas se hicieron con kits comerciales específicos en un analizador bioquímico semiautomático. Los resultados fueron: glucosa 37.04, 41.19 y 30.07 mg/dl; proteínas totales 1.65, 1.93 y 1.79 g/dl; albúmina 1.14, 1.42 y 1.31 g/dl; colesterol 15.82, 18.3 y 11.08 mg/dl; triglicéridos 11.3, 12.35 y 10.78 mg/dl para los estadios de crecimiento, estático y regresión, respectivamente, sin diferencias significativas entre estadios, con excepción de la glucosa. La concentración de glucosa fue menor en el estadio de regresión en comparación con el estadio de estática (p<0.05).

ABSTRACT The study aimed to characterize the follicular emergence and determine the repeatability of the follicular wave in alpacas. For this, 34 female alpacas were selected and induced to ovulation by parenteral application of buserelin acetate (day 0). Two days after the hormonal stimulation, individual follicular development was recorded by counting the total number of antral follicles >3 mm in both ovaries. For this, an inter-daily transrectal ultrasound follow-up was done for 60 days. It was determined that the emergence of the follicular wave occurred on day 4.5 ± 1.4 (range 2-8). The number of antral follicles (NAF) >3 mm during three consecutive follicular emergencies was 2.8 ± 1.3 (range 1-8). Despite having a great similarity (p>0.05) in the NAF >3 mm during three successive follicular emergencies there is a significant difference in the average number of emerging follicles between alpacas (p<0.05). The repeatability of the NFA >3 mm in the three follicular waves was 0.72. The interval between waves was 16.9 ± 3.9 days (range 8-24) and was identified as variable among animals, which showed a repeatability of 0.23. It is concluded that the number of emerging follicles >3 mm represents a highly repeatable characteristic in alpacas, but variable among animals; the interval between waves was also variable and a non-repeatable character.

RESUMEN El presente trabajo tuvo como objetivo caracterizar la emergencia folicular y determi- nar la repetibilidad de la onda folicular en alpacas. Para ello, 34 hembras alpacas fueron seleccionadas e inducidas a ovulación mediante la aplicación parenteral de acetato de buserelina (día 0). Dos días después del estímulo hormonal, se registró el desarrollo folicular individual a partir del conteo del número total de folículos antrales >3 mm en ambos ovarios. Para esto, se hizo un seguimiento ecográfico transrectal inter diario por 60 días. Se determinó que la emergencia de la onda folicular se produjo el día 4.5 ± 1.4 (rango 2-8). El número de folículos antrales (NFA) >3 mm durante tres emergencias foliculares consecutivas fue de 2.8 ± 1.3 (rango 1-8). A pesar de haber una gran similitud (p>0.05) en el NFA >3 mm durante tres emergencias foliculares sucesivas existe una diferencia significativa en el número promedio de folículos emergentes entre alpacas (p<0.05). La repetibilidad del NFA >3 mm en las tres ondas foliculares fue de 0.72. El intervalo entre ondas fue de 16.9 ± 3.9 días (rango 8-24) y se identificó como un carácter variable entre animales, el cual presentó una repetibilidad de 0.23. Se concluye que el número de folículos emergentes >3 mm representa una característica altamente repetible en alpacas, pero variable entre animales; siendo además el intervalo entre ondas un carácter variable y no repetible.