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泰国亚洲象()的精液特征与第二次人工授精成功

Semen characteristics and second successful artificial insemination of Asian elephant () in Thailand.

作者信息

Thongphakdee Ampika, Kiatsomboon Supalak, Noimoon Sakhon, Kongprom Urarikha, Boonorana Itti, Karoon Santhita, Thawnern Jedsada, Sakulthai Apichaya, Sombutputorn Petthisak, Sukmak Manakorn, Punkong Chainarong, Thongtip Nikorn

机构信息

Wildlife Reproductive Innovation Center, Conservation and Research Institute, Zoological Park Organization of Thailand under the Royal Patronage of H.M. the King, Chonburi 20110, Thailand.

Khao Kheow Open Zoo, Zoological Park Organization of Thailand under the Royal Patronage of H.M. the King, Chonburi 20110, Thailand.

出版信息

Vet World. 2022 May;15(5):1246-1255. doi: 10.14202/vetworld.2022.1246-1255. Epub 2022 May 22.

DOI:10.14202/vetworld.2022.1246-1255
PMID:35765477
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9210840/
Abstract

BACKGROUND AND AIM

As the number of wild Asian elephants () continues to decline, maintaining healthy populations under human care is vital. Male fertility assessment is essential for understanding the reproductive status, which can help to uncover underlying problems and improve the rate of pregnancy success. The objectives of this study in Asian elephants were as follows: (1) To investigate the semen characteristics; (2) to compare the relative seminal vesicle size and semen characteristics; (3) to compare the semen characteristics between good-motile (>60% progressive motility) and poor-motile (<60% progressive motility) ejaculates; and (4) to investigate the pregnancy success rate after artificial insemination (AI) with combined chilled and frozen semen.

MATERIALS AND METHODS

In total, 153 ejaculates were collected by manual rectal stimulation from 25 bulls. The volume, pH, sperm concentration, progressive motility, viability, morphology, and membrane integrity were investigated in each ejaculate. Assessment of accessory sex glands was conducted using transrectal ultrasonography to compare the relative seminal vesicle size and semen characteristics, and the bulls were divided into two groups according to the size of the ampulla (<7 or ≥7 cm). For the comparison of good and poor-motile ejaculates and semen characteristics, the samples were divided into two groups: Good-motile (>60% progressive motility) and poor-motile (<60% progressive motility) ejaculates. Semen ejaculates for AI were collected from three bulls. The estrous cycles of four females were monitored using an enzyme immunoassay. Seven AI attempts were conducted using frozen and/or chilled semen by endoscopic visualization. AI was repeated 1 day before the luteinizing hormone surge, on the day of the surge, and 1 day after the surge. Pregnancy was confirmed by monitoring the serum progesterone profile and the abdomen and mammary glands changes.

RESULTS

From 153 ejaculates, the mean±standard error values of progressive motility, semen volume, sperm concentration, pH, and viability were 40.18%±2.28%, 40.94±3.86 mL, 1,205.58±62.26×10 sperm/mL, 7.50±0.10, and 56.17%±1.96%, respectively. Comparing ampulla size and semen characteristics revealed that the bulls with ampullae of ≥7 cm yielded significantly larger volume ejaculates. However, there were no significant differences in sperm motility and concentration. The comparison of semen characteristics between good- and poor-motile ejaculates revealed that the former had significantly higher pH, viability, normal acrosomes, intact membranes, and normal head and tail morphology but often had a significantly lower volume and sperm concentration. From seven AI attempts in four females, one female had a confirmed pregnancy (14.3% pregnancy rate), and delivered a healthy live female baby weighing 128 kg at 21 months and 12 days of gestation. The baby is now 3 years old and in a healthy condition, with normally developing growth and behavior.

CONCLUSION

The semen characteristics of Asian elephants can be used as the baseline reference for further applications. The ampullae size indicates semen quantity but not quality. Our success in producing an elephant calf from AI using frozen and chilled semen demonstrated that AI can be used as an alternative approach for the breeding management of Asian elephants. However, the semen of Asian elephants is of poor quality, especially in terms of membrane integrity; thus, the improvement in semen quality through intensive and careful management of elephant health and fertility remains a challenge for the future. Furthermore, a sperm bank should be established to develop sperm cryopreservation, which will be invaluable for improving the genetic diversity of the Asian elephant.

摘要

背景与目的

随着野生亚洲象数量持续减少,在人工饲养条件下维持健康种群至关重要。雄性生育能力评估对于了解生殖状况至关重要,有助于发现潜在问题并提高怀孕成功率。本研究针对亚洲象的目标如下:(1)研究精液特征;(2)比较精囊相对大小与精液特征;(3)比较活力良好(>60% 渐进性运动)与活力较差(<60% 渐进性运动)射精的精液特征;(4)研究使用冷藏和冷冻精液联合人工授精(AI)后的怀孕成功率。

材料与方法

通过直肠手动刺激从25头雄象共采集了153份射精样本。对每份射精样本的体积、pH值、精子浓度、渐进性运动、活力、形态和膜完整性进行了研究。使用经直肠超声对附属性腺进行评估,以比较精囊相对大小与精液特征,并根据壶腹大小(<7或≥7 cm)将雄象分为两组。为比较活力良好与较差射精的精液特征,样本分为两组:活力良好(>60% 渐进性运动)和活力较差(<60% 渐进性运动)射精样本。从三头雄象采集用于人工授精的精液样本。使用酶免疫测定法监测四只雌象的发情周期。通过内镜可视化,使用冷冻和/或冷藏精液进行了七次人工授精尝试。在促黄体生成素激增前1天、激增当天和激增后1天重复进行人工授精。通过监测血清孕酮水平以及腹部和乳腺变化来确认怀孕。

结果

从153份射精样本来看,渐进性运动、精液体积、精子浓度、pH值和活力的平均值±标准误差分别为40.18%±2.28%、40.94±3.86 mL、1,205.58±62.26×10⁶精子/mL、7.50±0.10和56.17%±1.96%。比较壶腹大小与精液特征发现,壶腹≥7 cm的雄象射精量显著更大。然而,精子活力和浓度无显著差异。活力良好与较差射精的精液特征比较显示,前者的pH值、活力、顶体正常、膜完整以及头部和尾部形态正常的比例显著更高,但体积和精子浓度通常显著更低。在四只雌象的七次人工授精尝试中,一只雌象确认怀孕(怀孕率14.3%),并在妊娠21个月零12天时产下一头健康的雌性幼象,体重128 kg。这头幼象现在3岁,健康状况良好,生长和行为发育正常。

结论

亚洲象的精液特征可作为进一步应用的基线参考。壶腹大小表明精液数量而非质量。我们使用冷冻和冷藏精液通过人工授精成功产下一头象犊,证明人工授精可作为亚洲象繁殖管理的替代方法。然而,亚洲象的精液质量较差,尤其是在膜完整性方面;因此,通过精心管理大象健康和生育能力来提高精液质量仍是未来的一项挑战。此外,应建立精子库以发展精子冷冻保存技术,这对于提高亚洲象的遗传多样性将具有重要价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f05/9210840/d46698e056fa/Vetworld-15-1246-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f05/9210840/6050b3211757/Vetworld-15-1246-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f05/9210840/5fca9abfedda/Vetworld-15-1246-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f05/9210840/d338ae70de0e/Vetworld-15-1246-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f05/9210840/d46698e056fa/Vetworld-15-1246-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f05/9210840/6050b3211757/Vetworld-15-1246-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f05/9210840/5fca9abfedda/Vetworld-15-1246-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f05/9210840/d338ae70de0e/Vetworld-15-1246-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f05/9210840/d46698e056fa/Vetworld-15-1246-g004.jpg

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