Morphology of Day-7 Bovine Demi-Embryos and their Post-transfer Viability*

Day 7 bovine embryos were microsurgically bisected and replaced into surrogate zonae pellucidae. They were fixed immediately after bisection and at various intervals of in vitro incubation at 35 °C in modified Dulbecco's medium. At the light microscopical level, the bisected embryos restored the prebisection morphology within 30 min. after splitting. The electron microscopy confirmed these findings, suggesting that day 7 bovine demi-embryos for transfer purposes, should be cultured for 30 min before morphologically evaluated. Eleven pairs of bisected day 7 bovine embryos were transferred to 11 synchronized heifers. The recipient heifers were slaughtered at day 15, and the recovered embryos evaluated. Nine of the demi-embryos developed to morphologically, normal spherical to elongated, embryos.     

Reproductive performance of crossbred and purebred male rabbits

The effect of buck genetic type and crossbreeding parameters on fertility and prolificacy were estimated using two rabbit sire lines and their reciprocal crosses. The relationship between the reproductive performance of inseminated multiparous does and several semen quality traits was also investigated. The semen characteristics evaluated were: pH (pH), mass and individual motility (MM, IM), percentage of viable spermatozoa (Vi), spermatozoa with normal apical ridge (NAR), normal spermatozoa (NSP), spermatozoa with morphological abnormalities of head (HAP), neck-midpiece (NAP), and tail (TAP), spermatozoa with the presence of proximal (PD) and distal (DD) cytoplasmic droplets.

Fertility was analysed as a continuous trait (kindling rate) or as a binary trait (success or failure of kindling). In the first case, the analysis was performed using GLM procedures of SAS v.8 according to a model that included the fixed factors of buck genetic type, number of ejaculates per pool and week of insemination. In the second case, fertility was analysed using GENMOD procedures of SAS v.8 according to a mixed model including the same fixed factors as before plus the physiological status of the does and the permanent random effect of female. Number of kits born alive and number of stillborn were analysed with MIXED procedures of SAS v.8 with the same model used for the analysis of fertility as a binary trait. Estimates of the estimable functions of crossbreeding genetic parameters of the lines were obtained from the solutions of the corresponding models by generalized least squares using GLM, GENMOD and MIXED procedures. Crossbreeding parameters were estimated according to the model of Dickerson. A linear regression was used to determine the relationship between fertility and litter size and the semen characteristics evaluated.

Significant differences in fertility were observed among buck genetic types, which were favourable to type R. Differences between lines in maternal genetic effects were relevant and favourable to type R for fertility. Individual heterosis was important but unfavourable for fertility.

A slight correlation was obtained between all semen quality traits and fertility and prolificacy. Two multiple models were found for fertility, including NAP, IM, NSP, buck genetic type and Vi in one model or NAR in other model. Individual motility had an important positive effect, while NAP had a small negative effect. When MM, TAP and buck genetic type were included in a multiple model for the number of kits born alive, both MM and TAP had significant small effects. Individual motility and DD appeared to be related to number of kits stillborn, but only DD had a significant although negligible effect.     

Low temperature storage and in vitro germination of cherimoya (Annona cherimola Mill.) pollen

Due to the protogynous dichogamy of cherimoya and to the absence of proper pollinating vectors, hand-pollination with fresh pollen is a common practice for cherimoya commercial production. In order to optimize the process of hand-pollination, in this work we have studied the conservation of cherimoya pollen at −20, −80 and −196 °C for up to 3 months. In vitro pollen germination of fresh pollen was 57.1% and it was progressively reduced with conservation time at the three temperatures studied reaching a minimum after 3 months of storage of 10.4%, 14.2% and 13.6% at −20, −80 and −196 °C, respectively. Differences in germination among temperatures were only significant during the first 2 weeks of storage. Field pollinations with pollen stored for up to 3 months at the three temperatures show no yield differences compared to pollinations performed with fresh pollen. The results indicate that pollen collected and stored at sub-zero temperatures at the beginning of the cherimoya blooming season can be used along the whole blooming season avoiding the need of collecting fresh pollen daily.     

Effects of α-tocopherol and Ascorbic Acid on Equine Semen Quality after Cryopreservation

This study investigated the effects of ascorbic acid and α-tocopherol supplementation on semen quality parameters of equine thawed-frozen semen. Semen was divided in seven different treatments in a final concentration of 100 × 10⁶ sperm/mL by using Gent extender containing no supplements (control) and the following supplements with three different concentrations: α-tocopherol (0.5, 1, and 2 mM) and ascorbic acid (0.45, 0.9, and 1.8 g/L). After thawing, all samples were maintained at 37°C, while analyses were performed at 0, 60, and 120 minutes. Evaluation of viability and acrosome status (using Pisum sativum agglutinin conjugated to fluorescein isothiocyanate and propidium iodide), mitochondrial membrane potential (5,5′,6,6′-tetrachloro-1,1′,3,3′tetraethylbenzimidazolyl carbocyanine iodine [JC-1]), membrane lipid peroxidation (LPO; C₁₁-BODIPY^581/591), and stability of the plasmatic membrane (merocyanine 540 and Yo-Pro-1) of each sample was determined by flow cytometry. Relative to the control group, supplementation with α-tocopherol improved (P ≤ .05) postthaw membrane LPO, yet the higher concentrations of ascorbic acid (0.9 and 1.8 g/L, respectively) showed a negative effect on membrane LPO. Neither antioxidant significantly increased (P > .05) the acrosome integrity and mitochondrial membrane potential of frozen-thawed spermatozoa, although supplementation with α-tocopherol and ascorbic acid (0.9 and 1.8 g/L, respectively) had a positive effect on membrane integrity and stability (P ≤ .05). For all semen parameters, the lower concentration of ascorbic acid (0.45 g/L) did not show significant differences (P > .05) compared with the control. In conclusion, α-tocopherol seems to be an efficient antioxidant for reducing the oxidative stress provoked by cryopreservation, decreasing lipid peroxidation on equine spermatozoa.     

贵州钙果发展前景、存在的问题及对策

从钙果的生物学特性、适应性、开发利用前景、贵州钙果发展优势等方面进行综合分析,探讨贵州钙果产业发展的对策措施。     

小核盘菌保藏过程中活力和致病力影响因素研究

研究了小核盘菌保藏过程中活力和致病力影响因素,结果表明：接种后培养6 d的小核盘菌活力（以该菌在PDA平板上培养形成的菌落直径表示）和致病力（以该菌在蒲公英叶片上形成的菌斑直径表示）最大,分别为66.2 mm和52.9 mm。保藏期间小核盘菌的活力和致病力均随时间呈下降趋势。保藏90 d后,培养8 d的小核盘菌的活力和致病力最大,分别为31.7 mm和17.9 mm;培养4 d的小核盘菌的活力和致病力最小,分别为6.3 mm和2.6 mm。保藏期间低水活度有利于小核盘菌活力和致病力的保持,活力和致病力均随水活度的降低而增加。当水活度低于0.432时,活力和致病力不再增加,趋于稳定。低温有利于小核盘菌活力和致病力的保持, 25 ℃下保藏90 d的小核盘菌失去了全部活力和致病力,而-16 ℃下小核盘菌的活力和致病力仅分别下降25.8 %和 13.7%。     

无籽瓯柑无核原因的研究

无籽瓯柑Citrus suavissima 'Seedless'是浙江省新选育的柑橘良种.从花粉的角度对无籽瓯柑、普通瓯柑Citrus suavissima,处红柚Citrus grandis 'Chuhong'和翡翠柚的Citrus grandis的花粉进行了花粉形态观测、花粉量测定、花粉生活力测定及花粉发芽试验等比较研究,试图探索无籽瓯柑无籽的原因.结果表明,4个品种的花药数相近,但无籽瓯柑的花药上罕见自然散发的花粉;无籽瓯柑和普通瓯柑的花粉量接近,两者无明显差异;翡翠柚的花粉量是4个品种中数量最多的,大约是瓯柑的4倍和处红柚的2倍;普通瓯柑花粉的生活力与翡翠柚和处红柚的花粉生活力相当,无籽瓯柑花粉的生活力略低;无籽瓯柑在培养基上不发芽,而普通瓯柑、处红柚和翡翠柚的花粉发芽率在蔗糖质量分数为100 g·kg-1和150 g·kg-1的培养基上接近.无籽瓯柑无籽是因为其花药上罕有自然散发的花粉以及花粉难发芽造成的.图1表3参13     

花龄对蓝莓柱头可授性及花粉活力的影响

以 南高丛蓝莓‘蓝雨’为试材,研究了花龄对柱头可授性和花粉活力的影响。结果表明：‘蓝雨’蓝莓雌雄性器官的育性具有同步性,有利于自然授粉结实。从花后第1d-5d花粉活力都较高,特别是花后第2d-4d,花粉活力保持在50%以上,是人工辅助授粉花粉的良好收集期;从花后第2d-4d柱头分泌黏液较多,此期柱头具有强可授性,是开展人工辅助授粉的最佳时期。     

牛冷冻精液质量评估研究进展

人工授精(AI)是第一代动物繁殖生物技术,最近几十年以来,研究人员已经探索出了许多评估冷冻-解冻后精液品质的新技术。由于精子质膜在低渗条件下的功能状态、离子分布及浓度和各种细胞器的功能都与冷冻解冻后精子的存活能力相关,冻精质量的评定也从最初的形态和主观活率分析,发展到更精确的染色质、质膜完整性及分解代谢活性的分子变化分析。但是,在实际应用当中,许多方法花费很大,况且单个的评估参数都与动物精子实际的受精力相关性不强。因此,要提高牛冻精质量的预言强度,应该将这些独立的参数结合起来,利用多元回归分析,建立一套更完美更精确的评估体系。     

不同稀释液生理参数对关中奶山羊精液 保存及冷冻效果的研究

[目的]探讨了不同pH值、渗透压的稀释液对关中奶山羊冻精品质的影响,以确定关中奶山羊精液最佳保存与冷冻效果。[方法]通过显微检测在不同pH值、渗透压的稀释液处理对降温前、平衡后与解冻后的精子的活率、复苏率、精子顶体完整率、精子畸形率及精子冻后存活时间等指标进行评估。[结果],关中奶山羊精液稀释液pH值为7.3时,降温前、平衡后与解冻后的精子活率最高,分别达到0.742±0.011%、0.645±0.011%、0.489±0.012%;冻后活率、复苏率及解冻后顶体完整率分别达到0.431±0.009%、63.1±0.427%、65.7±2.018%;精子存活时间在降温前、平衡后及解冻后分别达到12.50±0.141h、11.96±0.186h、11.61±0.753h,冻后 8 h 活率0.52±0.014%;当渗透压为Δ0.754℃时,其精子的保护效果最佳,降温前、平衡后与解冻后的精子活率分别达到0.752±0.045%、0.671±0.024%、0.649±0.036%;冻后活率、复苏率及解冻后顶体完整率分别达到0.481±0.019%、63.6±0.377%、69.9±1.068%;精子存活时间在降温前、平衡后及解冻后分别达到11.82±0.165h、11.94±0.145h、11.70±0.753h,冻后 8 h 活率达到0.43±0.033%。[结论]关中奶山羊精液稀释液最合适的pH值是7.3,最佳渗透压为Δ0.754℃,这一结果对奶山羊稀释液配方优化提供了一定的理论指导。