Biotecnología de la Reproducción, Estación Experimental Agropecuaria Balcarce, Instituto Nacional de Tecnología Agropecuaria, 7620 Balcarce, Argentina.
Anim Reprod Sci. 2010 Jun;119(3-4):322-8. doi: 10.1016/j.anireprosci.2010.02.001. Epub 2010 Feb 6.
Two experiments were conducted to determine the effect of estradiol benzoate (EB) and intravaginal progestagen treatment on ovarian follicular dynamics and superstimulatory response in eCG-treated llamas. The purpose of Experiment 1 was to evaluate the effect of EB and progestagen treatment starting at different phases of dominant follicle (DF1) development on regression pattern and subsequent follicle wave emergence (WE2) in lactating and non-lactating llamas. Early lactating (n=24, 30+/-4 days postpartum) and non-lactating (n=24) females were assigned in equal numbers (n=8) to one of three groups according to the phase of DF1 (growing, static or regressing) determined by ultrasonography from day -3 to day 0. At day 0, llamas received an intravaginal sponge (MPA, 150 mg) and 5mg of MPA (i.m.). Half of the females (n=4) of each group were injected with 2mg (i.m.) of EB and half were not (control group). A 2 x 2 x 3 (lactational status, EB treatment and follicular phases) factorial design was used. Each sponge was removed 8 days later. Ovaries were monitored from day 0 to day 12. Daily blood samples were taken to determine 17beta-estradiol (E(2)) profiles from day 0 to day 8. The DF1 regression pattern was not affected (P>0.05) by the phase of follicle wave at the start of the treatment or any interactions among main effects. Follicle wave emergence in EB-treated llamas was delayed (P<0.05) by 2.3 days compared with non-treated llamas. Following EB treatment, plasma concentrations of E(2) were greater (P<0.05) from day 1 to day 5 in the treated than in non-treated females, but not from day 6 onward (P>0.05). Experiment 2 was designed to evaluate the effect of this treatment on the ovarian superovulatory response and embryo yield following eCG treatment administered on day of follicular wave emergence as determined in the Experiment 1. The same lactating (n=18, 61+/-4 days postpartum) and non-lactating (n=18) llamas at random stages of follicle wave were treated as those in Experiment 1 and received 1200IU of eCG at the time of WE2 (EB-treated=day 6.5 and non-treated=day 4.5). Llamas were mated 5 days after sponge withdrawal. A second mating was allowed 24h later. Embryos were collected between 7 and 8 days after the first mating and blood samples were taken to determine progesterone (P(4)) concentrations. The mean number of follicles on day of mating and the number of CL on day of embryo collection were not affected by lactational status, EB treatment or their interactions (P>0.05). Ovulation rate and mean (+/-SEM) number of recovered embryos for EB treatment group (67.5% and 2.4+/-0.4) were greater (P<0.05) than for no EB treatment (51.1% and 1.1+/-0.4). Plasma P(4) concentrations and number of CL per llama were correlated (r=0.49; P=0.014). In conclusion, progestagen plus EB treatment facilitates the prediction of the emergence of a new follicular wave approximately 6 days after treatment and resulted in a higher ovulation rate and embryo production in ovarian superstimulated llamas regardless of lactational status.
两项实验旨在确定苯甲酸雌二醇(EB)和阴道孕激素处理对 eCG 处理的美洲驼卵巢卵泡动态和超刺激反应的影响。实验 1 的目的是评估在优势卵泡(DF1)发育的不同阶段开始的 EB 和孕激素处理对泌乳和非泌乳美洲驼的卵泡波出现(WE2)的回归模式和随后的卵泡波出现的影响。根据超声检查从第-3 天到第 0 天确定的 DF1 阶段(生长、静止或退化),将早期泌乳(n=24,产后 30+/-4 天)和非泌乳(n=24)雌性等数量分配到三个组之一(n=8)。在第 0 天,美洲驼接受阴道海绵(MPA,150mg)和 5mg MPA(肌肉注射)。每组的一半(n=4)雌性接受 2mg(肌肉注射)EB,另一半为对照组。采用 2 x 2 x 3(泌乳状态、EB 处理和卵泡阶段)因子设计。8 天后取出每个海绵。从第 0 天到第 12 天监测卵巢。从第 0 天到第 8 天每天采集血液样本以确定 17β-雌二醇(E2)谱。DF1 退化模式不受处理开始时卵泡波阶段或主要效应之间任何相互作用的影响(P>0.05)。EB 处理的美洲驼卵泡波出现延迟(P<0.05),比未处理的美洲驼延迟了 2.3 天。EB 处理后,处理组的血浆 E2 浓度从第 1 天到第 5 天高于未处理组(P<0.05),但从第 6 天起(P>0.05)。实验 2 旨在评估这种处理对根据实验 1 中确定的卵泡波出现日期给予 eCG 处理后卵巢超刺激反应和胚胎产量的影响。随机处于卵泡波不同阶段的相同泌乳(n=18,产后 61+/-4 天)和非泌乳(n=18)美洲驼接受与实验 1 相同的处理,并在 WE2 时接受 1200IU 的 eCG(EB 处理=第 6.5 天,非处理=第 4.5 天)。海绵取出后 5 天配种。允许 24 小时后进行第二次配种。第一次配种后 7-8 天收集胚胎,并采集血液以确定孕酮(P4)浓度。配种当天的平均卵泡数和胚胎收集当天的 CL 数不受泌乳状态、EB 处理或它们之间的相互作用的影响(P>0.05)。EB 处理组的排卵率和平均(+/-SEM)回收胚胎数(67.5%和 2.4+/-0.4)高于无 EB 处理组(51.1%和 1.1+/-0.4)(P<0.05)。血浆 P4 浓度和每只美洲驼的 CL 数呈正相关(r=0.49;P=0.014)。总之,孕激素加 EB 处理有助于预测大约 6 天后新卵泡波的出现,并导致泌乳或非泌乳超刺激美洲驼的排卵率和胚胎产量更高。