CN103126728B - Determination method of normal or delayed ovulation in pandas - Google Patents

Determination method of normal or delayed ovulation in pandas Download PDF

Info

Publication number
CN103126728B
CN103126728B CN 201310070672 CN201310070672A CN103126728B CN 103126728 B CN103126728 B CN 103126728B CN 201310070672 CN201310070672 CN 201310070672 CN 201310070672 A CN201310070672 A CN 201310070672A CN 103126728 B CN103126728 B CN 103126728B
Authority
CN
Grant status
Grant
Patent type
Prior art keywords
panda
peak
time
female
estradiol
Prior art date
Application number
CN 201310070672
Other languages
Chinese (zh)
Other versions
CN103126728A (en )
Inventor
侯蓉
张志和
罗娌
黄河
王成东
兰景超
黄祥明
Original Assignee
成都大熊猫繁育研究基地
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Grant date

Links

Abstract

本发明公开了一种大熊猫正常或延迟排卵的确定方法,其以雌性大熊猫雌二醇真实峰值及其出现时刻为基础准确地检测出了雌性大熊猫LH峰值及其出现时刻,然后依据LH峰值时刻与雌二醇真实峰值时刻的时间间隔和雌性大熊猫是否出现发情高峰行为等条件共同确定雌性大熊猫正常或延迟排卵现象,准确地掌握了雌性大熊猫的排卵规律,为雌性大熊猫适时配种提供了适当时机,极大地促进了对濒危动物大熊猫的保护。 The present invention discloses a method for determining the normal or delay ovulation pandas, giant pandas female estradiol which is true peak appears in time and accurately detected based on the peak of LH and female panda appearance time, and then based LH time peak time and peak estradiol real time interval and whether the conditions for female panda estrus peak behavior appear jointly determine the female panda normal phenomenon or delay ovulation, ovulation accurately grasp the laws of female giant panda, is timely female panda breeding provides the appropriate time, which greatly contributed to the protection of endangered giant pandas.

Description

大熊猫正常或延迟排卵的确定方法 The method of determining the normal or delay ovulation panda

技术领域 FIELD

[0001] 本发明涉及一种用于圈养动物排卵情况的判别方法,特别是涉及一种熊猫正常或延迟排卵的确定方法。 [0001] The present invention relates to a method for determining ovulation for captive animals, in particular, relates to a method for determining the normal or delay ovulation panda.

背景技术 Background technique

[0002] 大熊猫自然交配成功率目前依然不够理想,确定其确切的适时配种时机是提高自然交配成功率的关键技术之一也是主要的技术障碍。 [0002] giant pandas mate naturally present success rate is still not ideal, timely determine the exact timing of breeding is to improve one of the key technologies is also a major technical hurdle natural mating success rate. 大熊猫自然交配比例偏低一直以来都是影响大熊猫成功繁殖的重要因素,据统计,自1936年至1989年间,在国、内外41个动物园、研究机构及自然保护区内累计圈养的大熊猫数量超过240只,但其中具有自然交配能力的雄性大熊猫个体却较少;胡锦矗.1990.大熊猫生物学研究与进展.成都:四川科学技术出版社.P316,统计出这一时期共有8只雄性大熊猫具备自然交配能力,仅占圈养雄性大熊猫总数的7%。 Pandas mate naturally low proportion has always been an important factor in the success of giant panda breeding influence, according to statistics, from 1936 to 1989, in the country, inside and outside the 41 zoos, research institutions and captive giant panda nature reserve accumulation the number more than 240, but the individual has a natural mating male giant panda has less ability; HU Jin-chu. 1990 Chengdu Giant panda Research and Development Biology: Sichuan Science and technology Press .P316, the statistics in this period a total of eight male giant pandas mate naturally have the ability, males accounted for only 7% of the total number of captive giant pandas. 冯文和、张安居等.1997.不同配种方式对大熊猫产仔效果的影响.四川大学学报(自然科学版)34:43-47,对1980~1993年圈养大熊猫155次不同繁殖方式的繁殖结果进行了统计,发现自然交配的个体数合计为57只次,仅占配种个体的36.8%(57/177)。 .. 1997, Fong Man and influence, Zhang homes, such as different ways of giant panda breeding farrowing effect of Sichuan University (Natural Science) 34: 43-47, 1980 to 1993, 155 captive giant panda breeding in different ways reproduction results were statistically and found that the total number of individual natural mating is 57 times, accounting for only 36.8% of breeding individuals (57/177). 上世纪90年代末期以来,通过增加发情期雌雄个体信息交流,延长母兽断奶时间等措施,有效提高了大熊猫自然交配比例,大约60%的成年雄性大熊猫具备自然交配能力。 Since the late 1990s, the exchange of information by increasing the individual male and female estrus, prolonged weaning time the mother animal and other measures to improve the proportion of giant pandas mate naturally, about 60% of adult male giant pandas mate naturally have the ability.

[0003] 近年来,由于加强大熊猫种群质量建设,适当限制了部分自然交配能力较强雄性个体的使用,并严格按照种群遗传管理原则制定雌雄个体配对计划,进一步增加了自然交配成功的难度,大熊猫目前自然交配的比例有所下降,比例依然偏低。 [0003] In recent years, strengthen the construction of the giant panda population quality, appropriate restrictions on the use of strong male individuals part of the natural mating ability, and strictly in accordance with the development plan paired male and female individuals genetic management principles, further increasing the difficulty of natural mating success, At present the proportion of giant pandas mate naturally declined, the proportion is still low.

[0004] 由于大熊猫一年中具有独特的生理习性,一年中只有I~3天接受自然交配,个别个体甚至仅仅几小时。 [0004] Since the panda has a unique physiological habits of the year, the only year I ~ 3 on Days natural mating, only a few individuals or even a few hours. 因此,确定适时配种时间是提高大熊猫自然交配成功率的关键技术之一O Therefore, timely determine the breeding time is a key technology to improve the success rate of giant pandas mate naturally O

[0005] 人工授精时机的把握存在观点存在争议:除了自然交配以外,人工授精是大熊猫繁殖的另一个重要手段。 Grasp [0005] artificial insemination opportunity exists controversial point of view: in addition to natural mating, artificial insemination is another important means of Giant Panda Breeding. 但是目前大熊猫人工授精技术尚存在以下问题:尽管多数情况下采用新鲜精液进行人工授精,但是大熊猫人工授精产仔率尚不够理想,健康适龄个体人工授精产仔率大约在40%-50%,且年度波动大,而大熊猫自然交配产仔率为60%-70%,表明目前大熊猫人工授精技术尚未成熟。 But now surviving giant panda artificial insemination techniques in the following issues: Although artificial insemination with fresh semen in most cases, but artificial insemination panda birth rate is still not ideal, healthy individuals age artificial insemination birth rate of about 40% -50% and annual fluctuations, and pandas mate naturally calving rate of 60% -70%, indicating that the current giant panda artificial insemination technology is not yet mature. 这可能与大熊猫适时人工授精时机存在争议有关。 This may be controversial about the timing and artificial insemination panda timely.

[0006]目前在大熊猫适时人工授精中,最主要的指标采用尿雌激素指标。 [0006] In the current giant panda artificial insemination in a timely manner, the most important indicators of the use of urinary estrogen targets. 但是对采用雌激素指标作为大熊猫适时配种的指标存在很大的争议,尚无统一观点。 But there is a big controversy on the index as an indicator of the use of estrogen timely giant panda breeding, there is no unified point of view.

[0007]目前普遍使用酶联免疫吸附试验检测大熊猫尿样雌激素。 [0007] current widespread use of enzyme-linked immunosorbent assay panda urine estrogen. 若以大熊猫尿中雌激素水平峰值当日作为O天,不同研究人员提出了不同的适时配种雌激素指标。 In terms of panda urine estrogen levels as the peak day O day, different researchers have proposed different indicators of timely breeding estrogen.

[0008] Hodges 等(Hodges JK, Bevan DJ, Celma M, Hearn JP, Jones DM, KleimanDG,Knight JA,Moore DM.1984.Aspects of the reproductive endocrinology of thefemale Giant Panda (Ailuropoda melanoleuca)in captivity with special referenceto the detection of ovulation and pregnancy.J.Zool.,Lond203:253 ~267.)根据3只大熊猫的研究结果提出适时配种时间可能在第I天、第2天。 [0008] Hodges et (Hodges JK, Bevan DJ, Celma M, Hearn JP, Jones DM, KleimanDG, Knight JA, Moore DM.1984.Aspects of the reproductive endocrinology of thefemale Giant Panda (Ailuropoda melanoleuca) in captivity with special referenceto the detection of ovulation and pregnancy.J.Zool, Lond203:.. 253 ~ 267) at the time may be made timely breeding day I, day 2 the results of three studies GIANT. [0009] 李复东、彭世媛、叶志勇、何光昕、黄祥明.1993年.雌性大熊猫发情期尿中17 β-雌二醇与孕酮水平的变化及其与配种的关系.兽类学报13 (3):166-171,提出适时配种时间应该在第O天和第I天。 [0009] Li Fudong, Peng Shiyuan, Ye Zhiyong, He Xin light, HUANG Xiangming. 1993. Relationship changes estradiol and progesterone levels in their urine and 17 β- breeding female panda estrus. Mammals Journal 13 (3) : 166-171, timely breeding time should be made at the O and day I.

[0010] 李学兵、黄祥明、李光汉、李绍昌、张福祥、余建秋、钟顺隆、宋云芳、陈红卫、唐飞、南西.莎卡娜.1994a.大熊猫发情期尿激素的ELISA测定与适时配种.成都国际大熊猫学术研讨会论文集.成都:四川科学技术出版社.247-250,认为适时配种时间应该从O天开始配种一直复配至尿中促黄体生成激素LH峰值出现为止。 [0010] Lixue Bing, HUANG Xiangming, Li Guanghan, Lishao Chang, Zhang Fuxiang, Yu Jianqiu, Zhong Shunlong, Songyun Fang, Chen Hongwei, Tang Fei, Nancy. Shaka Na .1994a. ELISA assay panda estrus hormone in urine and timely breeding. Chengdu Giant Panda International Academic Symposium Chengdu: Sichuan Science and technology Press .247-250, time should always be considered timely breeding complex from day O to start breeding urinary luteinizing hormone LH peak occurs.

[0011] 上面研究人员所提出的大熊猫适时人工授精时机,均是以尿雌激素峰值出现时间为参照,大熊猫适时人工授精时机均没有足够的成功人工授精的实例来支持其观点的正确性是最大的问题。 Artificial insemination panda timely opportunity to [0011] above the researchers proposed, are based on urinary estrogen peak time as a reference, the timing of artificial insemination of giant pandas are not timely enough examples of successful artificial insemination to support the correctness of their views It is the biggest problem.

[0012] 从目前已经报道的成功进行大熊猫人工授精实例而言,一方面对大熊猫适时人工授精时机也缺乏较为一致的看法,并且已经公开报道的人工授精的个体数量较少,从而难以充分支持其观点。 [0012] Examples of the giant panda from artificial insemination carried out in terms of success already reported, on the one hand of the giant panda timely artificial insemination timing is also a lack of a more consistent view, and a small number of individuals have been publicly reported artificial insemination, making it difficult to fully support their views.

[0013] 黄炎.2001年.使用冷藏精液进行大熊猫(Ailuropoda melanoleuca)人工授精.应用与环境生物学报-2001.7(6).-558-562,报道了一例大熊猫人工授精案例,认为大熊猫人工授精的最佳时间在雌兽尿液中的雌激素达到最大峰值后的48~72小时,而在之后同一作者又否定了之前提出的这一看法。 [0013] Huang Yan in 2001 using frozen semen giant pandas (Ailuropoda melanoleuca) artificial insemination. Applied and Environmental Biology -2001.7 (6) .- 558-562, reported cases of artificial insemination panda examples, consider the giant panda the best time artificial insemination of estrogen in females in the urine of 48 to 72 hours after the maximum peak, and after the same author has denied the views expressed before.

[0014] 黄炎.2006年.雌性大熊猫发情期雌酮的研究.四川农业大学学位论文,通过对2004~2005年度对中国保护大熊猫研究中心的26只次大熊猫尿液雌酮含量进行了检测,并且分析了自然交配与人工授精发生时间与成功产仔的关系,提出雌性大熊猫交配或人工授精的最佳时间是尿液雌酮含量达到峰值的当天,即O天进行。 [0014] Huang Yan 2006 study female panda estrus estrone. Dissertation, Sichuan Agricultural University, by the content of the ketone from 2004 to 2005 for the Giant Panda Protection Research Center of China's 26 giant panda female urine the detection and analysis of the relationship between natural mating and artificial insemination time of occurrence and success of the litter, the best time to put forward female panda mating or artificial insemination of urine estrone content reaches a peak day, namely O days. 2012年度黄炎等(2012) Relationship of the Estrogen Surge and Multiple Mates to Cub Paternityin the Giant Panda(Ailuropoda melanoleuca):1mplications for Optimal Timing ofCopulation or Artificial Insemination.Biol Reprod.2012Nov8; 87 (5): 112.对大熊猫适时配种时间又提出了不同看法。 2012 Annual Huang Yan, etc. (2012) Relationship of the Estrogen Surge and Multiple Mates to Cub Paternityin the Giant Panda (Ailuropoda melanoleuca): 1mplications for Optimal Timing ofCopulation or Artificial Insemination.Biol Reprod.2012Nov8; 87 (5): 112. large Panda timely breeding time and put forward different views. 对经自然交配与人工授精进行繁殖的19只产仔大熊猫的分析,暗示大熊猫适时配种时间(包括自然交配与人工授精)为雌激素峰值后第二天。 Analysis of 19 litter for breeding pandas through artificial insemination and natural mating, suggesting the giant panda breeding in due course of time (including natural mating and artificial insemination) after the estrogen peak the next day.

[0015] 喻述容等.2003年.繁殖期雌性大熊猫尿中激素变化的研究.畜牧兽医学报-2003.34(5).-452-456),对大熊猫适时配种时间又提出了不同的看法,在对39只大熊猫发情期尿液雌酮以及配种时机与产仔结果进行分析后提出,在尿液雌酮峰值下降12h内进行首次自然交配、人工授精或两者交替配种,受孕率达100% ;峰值下降12 — 24h内进行首次自然交配、人工授精或两者交替配种,受孕率达83.3% ;峰值下降24h后进行首次配种,受孕率为O。 [0015] Yushu Rong etc. In 2003, research on reproductive hormonal changes in urine of female panda. Animal Husbandry and Veterinary Science -2003.34 (5) .- 452-456), giant panda breeding time and timely put forward different views, in after one of the 39 giant panda breeding and calving results and the timing of estrus females were analyzed urine suggested that one peak decline in female urine within 12h for the first time natural mating, artificial insemination or alternating between the two mating, the pregnancy rate of 100% ; value decreased 12-- 24h for the first natural mating, artificial insemination, or alternating between the two mating, conception rate of 83.3%; for the first breeding, pregnancy rate after 24h decreased peak O.

[0016] 后两篇报道尽管报道的人工授精案例较多,并且通过自然交配以及人工授精时机与产仔成功的关系提出了适时配种时间,但是后两篇报道存在的最大问题在于两位作者均将人工授精与自然交配这两种不同的配种方式合并统计,而没有单独统计单纯人工授精时机与成功产仔的关系。 [0016] Although the two reports after reports of more cases of artificial insemination, and made a timely breeding time through natural mating and artificial insemination timing relationship with calving success, but the biggest problem is that the authors after two reports were the artificial insemination and natural mating of these two different ways of breeding consolidated statistics, but there is no statistical relationship purely artificial insemination timing and success of litter alone. 由于自然交配并不需要麻醉,对排卵的影响较小,而人工授精则需要在麻醉状态下进行,若人工授精时机不当,将对排卵产生较大的影响,因此将人工授精与自然交配这两种不同的配种方式合并统计后所提出的适时人工授精时机不能真实反映合理的适时人工授精时机,因此这两位作者提出的人工授精时机尚缺乏足够依据,并且所提出的适时人工授精时机也缺乏后续应用以及产仔率结果的报道来证明其正确性。 Since natural mating does not require anesthesia, less impact on ovulation and artificial insemination will need to be under anesthesia, if improper timing of artificial insemination, ovulation will have a greater impact, so the artificial insemination and natural mating two timely opportunity artificial insemination breeding different ways after the proposed merger statistics can not reflect a reasonable timely opportunity artificial insemination, artificial insemination and therefore the timing of these two authors proposed lack of sufficient basis, timely artificial insemination timing and lack of proposed farrowing rate and subsequent application reports the results to prove its correctness.

[0017] 综上所述,关于现有的大熊猫适时人工授精技术,目前提出的适时人工授精时间均以尿液雌激素峰值出现时间作为判断依据,并且所提出的适时人工授精时间指标差异较大。 [0017] In summary, with respect to the existing panda timely artificial insemination, artificial insemination in due course of time it is currently proposed urine estrogen peak time as the judgment basis, timely insemination time difference and the proposed indicators than Big. 若以雌激素峰值出现当日作为O天,提出的适时人工授精时间分别为:第一种观点是第I天、第2天(Hodges等.1984,黄炎等.2012);第二种观点是第O天和第I天(李复东等.1993);第三种观点是从O天开始配种一直复配至尿中LH峰值出现为止(李学兵等.1994);第四种观点是O天(黄炎.2006);第五种观点是若以雌激素峰值出现时刻作为O小时,适时配种时间为O~24小时,最好为O~12小时(喻述容.2003)。 In terms of estrogen hormone peak occurs as time of day O timely insemination day, were proposed: The first point is the first day I, day 2 (Hodges et. 1984, Huang Yan, etc. .2012); second view is the first O and day I (Li Fudong, etc., 1993); third view is days has been compounded from O to start breeding until the urinary LH peak (Lixue Bing etc. 1994); the fourth point is O day (yellow inflammation. 2006); the fifth point is the moment of occurrence In terms of estrogen as O peak hours, timely breeding time O ~ 24 hours, preferably O ~ 12 hours (Yu Shurong .2003). 总结起来,若以雌激素峰值当日作为O天,对于首次人工授精时间目前的观点均为O天或者第一天。 To sum up, if the hormone estrogen peak day as O day, for the first time artificial insemination current view are O day or the first day.

[0018] 另一方面,目前的研究结果表明若主要以雌激素指标作为大熊猫人工授精的主要参考指标是不足的。 [0018] On the other hand, the current findings suggest that if the main index to estrogen as the main reference index panda artificial insemination is inadequate. 目前大熊猫适时人工授精主要通过监测尿液雌激素变化规律来确定大熊猫适时人工授精时机。 Currently timely artificial insemination panda is primarily determined by the timing of artificial insemination panda timely monitoring of urine variation of estrogen. 如果以雌激素峰值出现时刻作为参考,选择适龄(6~20岁)、健康个体统计,比较产仔组与未产仔组人工授精时间差异(数据来自成都大熊猫繁育研究基地),结果见表1: If the time as a reference, select age (6 to 20 years old) to the estrogen peak, artificial insemination time statistical differences in healthy individuals, comparing calving group and not litter group (data from the Chengdu Research Base of Giant Panda Breeding), the results in Table 1:

[0019] 表1.人工授精个体产仔组与未产仔组人工授精时间差异比较结果 [0019] Table 1. Artificial insemination is not subject group and calving group farrowing time difference comparison result insemination

[0020] [0020]

Figure CN103126728BD00051

[0021] 从表1可以看出,若以雌激素峰值时刻作为人工授精的主要参考指标,产仔组与未产仔组人工授精时刻非常接近,均距雌激素峰值26小时左右。 [0021] As can be seen from Table 1, if the estrogen peak time as the primary reference index insemination, calving group and not very close to the time of artificial insemination group farrowing, from both estrogen peak around 26 hours. 采用独立样本T检验对人工授精个体产仔组与未产仔组人工授精时间差异进行比较,结果P=0.851,差异不显著。 Independent sample T-test group and artificial insemination insemination individual calving time difference did not farrowing groups were compared, P = 0.851, the difference was not significant. 该结果表明,若主要以雌激素指标作为适时配种的主要参考指标是不足的,不能解释为什么在相似的时机作人工授精,造成部分个体产仔而大部分个体未产仔的结果。 The results show that, if the main indicators of estrogen as the main reference indicator is the lack of timely breeding, artificial insemination do not explain why the similar time, resulting in part the result of individual farrowing and most individuals are not calving. 其产仔率仅仅为46.7%,显著低于自然交配大熊猫高达60%-70%产仔率水平。 Farrowing rate which is only 46.7%, significantly lower than the natural mating panda up to 60% -70% farrowing rate level.

[0022] 促黄体生成激素是促性腺激素的一种,简称为LH,它作用于成熟的卵胞,能引起排卵并生成黄体。 [0022] The luteinizing hormone is a gonadotrophin, abbreviated as LH, which acts on the mature follicles, can cause ovulation and corpus luteum generated. 目前关于大熊猫LH的研究鲜有报道。 LH's current research on giant pandas rarely reported.

[0023] LH是一种糖蛋白激素,在雌性哺乳动物,LH的主要作用是与垂体分泌卵泡刺激素FSH协同促进卵泡生长成熟,粒膜增生,参与内膜细胞合成雌激素,并可诱发排卵,促进黄体生成,是诱发排卵的主要因素之一。 [0023] LH is a glycoprotein hormone in a female mammal, the main role is to LH secretion of pituitary follicle-stimulating hormone FSH synergistic mature follicular growth, granulosa hyperplasia, endometrial cells involved in the synthetic estrogen, can induce ovulation , luteinizing promoting, it is a major factor inducing ovulation.

[0024] LH由α和β两个糖基化亚基以非共价键方式连接形成的异二聚体。 [0024] LH by the α and β subunits of two glycosylation joined to form a non-covalently heterodimer. 各动物的α亚基均相同或十分相似,无激素间特异性和种间特异性。 Each animal α subunits are the same or very similar, no inter-specific and inter-species specific hormone. β亚基则存在很大的差异,不但有激素间特异性,且还有种间特异性,是激素生物活性和免疫活性的决定因素,故将β亚基称为特异亚基,能与靶器官受体结合。 β subunit is very different, not only between the hormone-specific, and there are species specific, is the deciding factor Hormone and immunological activities, it is referred to as the β-subunit-specific subunits, to the target organ receptor binding. 据廖明娟(廖明娟,2003,大熊猫等濒危动物FSH/LH和GH/GHR基因的克隆及其表达研究浙江大学博士论文ρ41)比较大熊猫FSH和LH亚基基因和其它几种哺乳动物的相应序列的同源性结果,见表2: According Liaoming Juan (Cloning and Expression Liaoming Juan, 2003, the giant panda and other endangered animals FSH / LH and GH / GHR gene thesis research Zhejiang University ρ41) compare panda FSH and LH subunit gene and the corresponding sequence of several other mammals homology results, shown in Table 2:

[0025] 表2.大熊猫与其它物种三亚基编码区核苷酸序列及其氨基酸序列的同源性比较 [0025] Table 2. pandas other species triylidene coding region nucleotide sequence and its homology comparison of amino acid sequences

Figure CN103126728BD00061

[0027] 注:比较结果为2010年3月以前数据,部分结果尚未有显示为未有相关数据报道。 [0027] Note: The result of the comparison to March 2010 before the data, partial results have not been displayed as the absence of relevant data reports.

[0028] 表2结果表明:大熊猫FSH和LH亚基基因无论在核苷酸水平还在氨基酸水平与人、狗、牛、猪、大小鼠具有很高的同源性;在所有被比较的物种中,大熊猫LH基因与犬类的同源性最高,核苷酸与氨基酸水平分别达到92%。 [0028] The results in Table 2 show that: the giant panda subunit genes FSH and LH also whether the amino acid level, humans, dogs, cows, pigs, mice and rats with high homology at the nucleotide level; all are compared in species, pandas LH gene highest homology, the nucleotide level and the amino acid reaches 92% in dogs.

[0029] 曾国庆、蒋广泰、杨克勤等.血清和尿液中促黄体素、孕酮和17β_雌二醇含量的变化.动物学报,1990,36(1): 63-69,对大熊猫LH进行了初步研究,LH检测采用体外生物测定法,对4只次大熊猫血液以及尿液中LH含量及其变化进行了初步测定,只是对发情期尿液样品每天采集了一次,而血液样品全部在人工授精麻醉动物时进行,结果初步发现大熊猫在发情高峰期出现LH峰值,LH峰值出现在雌激素峰值后。 .. [0029] was the National Day, Jiangguang Tai, Yang Keqin such as serum and urine luteinizing hormone, progesterone and estradiol 17β_ changes in animal Sinica, 1990,36 (1): 63-69, giant panda We were studied LH, LH in vitro bioassay used to detect, for four giant panda blood and urine LH and content of a preliminary determination, just estrus urine samples were collected once a day, and a blood sample all the animals were anesthetized at the time of artificial insemination, panda result of preliminary findings appear in estrus peak of the LH peak, LH peak after the estrogen peak.

[0030] 林增云、朱忠勇、陈玉春.大熊猫发情排卵期尿中LH、Ε2内分泌的研究.成都:四川科学技术出版社.1991.66-70,采用了不同的测定方法(采用碘标人类促黄体生成激素hLH放射免疫测定法)对2只次大熊猫LH进行了测定,发情高峰期每天收集一份尿样,结果与曾国庆的结果有所差异,大熊猫雌激素E2和LH在发情高峰第二天或者第三天,同步达到最大峰值。 . [0030] Lin Zengyun, Zhuzhong Yong, Chen Yuchun urinary LH panda estrus ovulation, endocrine studies ∈ 2 Chengdu: Sichuan Science and Technology Press .1991.66-70, using a different measurement method (using 125I human luteinizing hLH hormone radioimmunoassay) of the two giant panda LH were measured, the peak of the rut every day to collect a urine sample, the results with the results of the National day was different, panda estrogen E2 and LH peak at estrus second day or the third day, the maximum peak synchronization.

[0031] 李学兵、黄祥明、李光汉等.大熊猫发情期尿激素的ELISA测定与适时配种.成都国际大熊猫保护学术研讨会论文集.成都:四川科学技术出版社.1994:247-250,采用酶联免疫测定法对6只9次大熊猫发情期尿液LH进行了检测,每天采集一份晨尿,LH测定试剂盒采用美国Tetcan公司提供,其结果与曾国庆的结果比较类似,发现大熊猫尿LH峰值出现在雌激素峰值稍后(当日、次日或第三日)。 . [0031] Lixue Bing, HUANG Xiangming, Li Guanghan ELISA assay and other panda estrus hormone in urine and timely breeding Proceedings of the International Symposium on the protection of giant pandas in Chengdu, Chengdu: Sichuan Science and Technology Press .1994: 247-250, using Enzyme-linked immunosorbent assay for six nine giant panda estrus urine LH were detected, a daily morning urine collection, LH assay kit provided by the United States Tetcan company, its results with the results of the National day was relatively similar, found large Panda urinary LH peak in (the same day, next day or third day) estrogen peak later.

[0032] Barbara S.Durrantj Nicole Ravi da, Thomas Spady et al.2006New technologies for the study of carnivore reproduction.Theriogenology, 60:1729-1736,首次釆用商品化狗血清LH测定试剂盒,釆用快速免疫色谱LH测定法(rapid immunochromatographic LH assay)对4只9次大熊猫尿液中LH含量及其变化进行了定性检测,发现尿LH阳性检测结果出现在雌激素峰值下降后24小时。 [0032] Barbara S.Durrantj Nicole Ravi da, Thomas Spady et al.2006New technologies for the study of carnivore reproduction.Theriogenology, 60: 1729-1736, preclude the first dog serum LH measured using a commercial kit, preclude the use of immunochromatographic LH assay (rapid immunochromatographic LH assay) for 4 panda 9 and content of LH in urine qualitative detection found positive urinary LH detection result appears in peak 24 hours after estrogen decline.

[0033] 综合上述研究结果,相对比较一致的结果是:若以雌激素峰值作为O天,大熊猫LH峰值出现在雌激素峰值后O~3日,但其确切时间不同研究结果有所不同。 [0033] From the research, the results are relatively consistent: In terms of estrogen peak days as O, giant panda LH peak at O ​​~ 3 days after the estrogen peak, but the results of different studies the exact time is different.

[0034] 上述大熊猫LH研究报道存在以下问题:首先检测个体数量少(2~6只),代表性差,其次采样频率低,难以获得大熊猫LH的准确变化曲线,由于通常LH峰持续时间较短,如果采样频率低会漏掉真实的LH峰,而上述研究采样多数为每天I次,而Barbara则是随机采样;其次,研究方法上定量检测方法采用生物学测定法,或者人的LH测定试剂进行放射免疫或酶联免疫测定,而定性测定则采用狗血液LH测定试剂盒。 Exists [0034] The studies reported LH panda following problems: First, a small number of individual detector (2 ~ 6), poor representation, followed by the sampling frequency is low, it is difficult to obtain an accurate curve LH pandas, since typically the duration than the LH surge short, if the sampling frequency is low miss real LH peak, while the majority of these studies sampled I per day, and Barbara is randomly sampled; second, quantitative detection method research using biological assays, or of measuring human LH Reagents for enzyme immunoassay or radioimmunoassay, and the measurement is qualitative assay kit using dog blood LH. 上述廖明娟等的研究结果表明,大熊猫LH的α和β两个糖基化亚基同源性与狗更高,而与人的相似程度较低。 The results showed that like the above-described Liaoming Juan, pandas LH α and β of two glycosylated subunits higher homology with the dog, while a lower degree of similarity to human. 大熊猫LH的α亚基与狗在核苷酸以及氨基酸水平的同源性达到95%和90%,而与人的只有84%和74%,大熊猫LH的β亚基与狗在核苷酸以及氨基酸水平的同源性达到92%,而与人只有84%和75%,因此大熊猫LH与狗的同源性高于与人的同源性。 The α subunits of LH panda with dog nucleotide and amino acid homology level of 95% and 90%, while only 84% human and 74%, of the β subunit of LH panda nucleoside with dog acid, and amino acid homology level of 92%, while only 84% human and 75%, so the dog LH panda homology homology higher than human. 因此理论上讲,如果利用其它物种LH测定试剂盒检测大熊猫LH,采用狗的优于人的。 Thus Theoretically, if the use of other species panda LH LH assay kit, using the dog than human. 但是目前采用狗LH测定试剂盒检测大熊猫LH仅仅只有定性检测研究而没有定量检测研究。 But now pandas LH kit with only the qualitative detection of quantitative research studies and not determined by a dog LH.

[0035] 王建辰.动物生殖调控.合肥:安徽科学技术出版社.1998,张嘉保.动物繁殖学.长春:吉林科学技术出版社.1999:96,发现LH排卵峰对于触发排卵起着关键性的作用,不同动物在血中出现LH与触发排卵的间隔时间不尽相同,但是在同一种动物其LH峰值时间与触发排卵的间隔时间几乎是一致的。 [0035] Wangjian Chen regulation of animal reproduction Hefei: Anhui Science and Technology Press .1998, Zhang Bao Animal Reproduction Changchun: Jilin Science and Technology Press .1999: 96, ovulation LH peak was found to play a key trigger ovulation effect, and different animals appears LH triggers ovulation interval vary in the blood, but at the same time an animal which is almost the same LH peak and trigger ovulation of interval. 因此利用LH指标作为大熊猫适时配种依据从理论上讲是可行的,但是目前没有任何相关研究以及应用报道,特别是部分大熊猫存在延迟排卵现象。 Therefore, the use of LH as an indicator of giant panda breeding timely basis theoretically possible, but there is no research and application reports, especially in some giant panda there is a delay ovulation phenomenon. 发明内容 SUMMARY

[0036] 本发明要解决的技术问题是确定大熊猫发情期尿样LH定量检测方法、揭示大熊猫发情期特别是发情高峰期LH变化规律以及与雌激素及发情行为的关系、确定为大熊猫适时配种提供依据的大熊猫正常或延迟排卵的确定方法。 [0036] The present invention to solve the technical problem is to determine the panda estrus urine LH quantitative detection method, especially LH reveal changes of the peak of the rut and the relationship between estrogen and the behavior of giant panda estrus estrus, identified as the giant panda timely provide the basis for breeding pandas normal method of determining ovulation or delay.

[0037] 为解决上述技术问题,本发明一种大熊猫正常或延迟排卵的确定方法,包括以下步骤: [0037] To solve the above problems, a method to determine a normal or delay ovulation panda the present invention, comprising the steps of:

[0038] (I)春季繁育期间,依照专利申请号201210191762.5所述,按时间要求对雌性大熊猫进行尿样收集,然后对尿样采用酶联免疫吸附试验检测出雌性大熊猫雌二醇真实峰值及其出现时间; [0038] during the spring breeding (the I), according to said patent application No. 201210191762.5, time required to collect urine female panda, and a urine sample by enzyme linked immunosorbent assay detects the true peak estradiol female panda and the time appears;

[0039] (2)以出现雌性大熊猫雌二醇真实峰值的时刻为时间起点O小时,采用酶联免疫吸附试验检测O小时到48小时内全部尿样的LH值,找出LH峰值及其出现时间;所述LH峰值为雌二醇真实峰值时刻后的最大LH值; [0039] The time (2) to appear female panda estradiol real time as a starting point O peak hours, enzyme-linked immunosorbent assay value O LH hours all the urine within 48 hours, and to identify peak LH time of occurrence; LH peak is the maximum value after the LH peak estradiol real time;

[0040] (3)依据雌二醇真实峰值时刻、LH峰值时刻与雌二醇真实峰值时刻的时间间隔、雌性大熊猫是否出现发情高峰行为等条件共同确定雌性大熊猫正常或延迟排卵现象;所述雌性大熊猫发情高峰行为是指雌性大熊猫出现翘尾、待配姿势、压尾反射、后退接近雄兽等现象,特别是待配姿势。 [0040] (3) based on estradiol real peak time, peak time interval LH peak and estradiol real time, whether the conditions for female panda estrus peak behavior appear jointly determine the normal female panda or delay ovulation phenomenon; the said female panda peak heat action refers to female panda carryover appears to be with the posture, the end pressure is reflected back and so close to the male animal, in particular with the posture to be.

[0041] 所述的一种大熊猫正常或延迟排卵的确定方法,当雌性大熊猫LH峰值时刻与雌二醇真实峰值时刻的时间间隔小于16小时、且在雌二醇真实峰值时刻后的30小时内雌性大熊猫出现发情高峰行为时,则可以确定雌性大熊猫为正常排卵个体。 One kind panda [0041] The normal method of determining or delay ovulation, the time when the peak time female panda LH and estradiol true peak time interval is less than 16 hours, and 30 after the true peak timing estradiol when female panda estrus peak behavior occurs within hours, it can be determined to be normal ovulation female panda individuals.

[0042] 所述的一种大熊猫正常或延迟排卵的确定方法,当雌性大熊猫LH峰值时刻与雌二醇真实峰值时刻的时间间隔大于16小时、且在雌二醇真实峰值时刻后的30小时内雌性大熊猫未出现发情高峰行为时,则可以确定雌性大熊猫为延迟排卵个体。 One kind panda [0042] The normal method of determining or delay ovulation, the time when the peak time female panda LH and estradiol true peak time interval is greater than 16 hours, and 30 after the true peak timing estradiol within hours of peak estrus female panda behavior does not appear, it can be determined to delay ovulation, a female panda individual.

[0043] 所述的任何一种大熊猫正常或延迟排卵的确定方法,其LH值采用犬促黄体生成激素试剂盒检测。 [0043] according to any one of the panda normal or delay ovulation method for determining which dog use of LH luteinizing hormone kit.

[0044] 在研究过程中,以出现大熊猫雌二醇真实峰值的时刻为时间起点O小时,采用酶联免疫吸附试验检测O小时到48小时内的全部尿样LH值;作出LH变化曲线,以LH的最大值作为LH峰值,以出现LH峰值的尿样排泄时间作为大熊猫LH峰值时刻,计算出大熊猫LH峰值时刻与雌二醇真实峰值时刻的时间间隔;采用SPSS统计软件及描述性数据频率分析方法分析了40只次大熊猫发情期尿LH峰值与雌二醇真实峰值的间隔时间(h),结果见下表3: At time [0044] During the study, estradiol panda to appear true peak time of starting O hours, with all the urine within 48 hours of LH enzyme-linked immunosorbent assay hours to O; made LH curve, LH LH maximum value as a peak, appears in urine excretion time of peak LH LH peak time as pandas, giant pandas calculate the time LH peak time and peak time interval real estradiol; SPSS statistical software and descriptive frequency analysis data were analyzed 40 times panda estrus interval estradiol real peak urinary LH peak (h), the results in Table 3 below:

[0045] 表3.大熊猫发情期尿LH峰值与雌二醇真实峰值间隔时间(h): [0045] Table 3. panda estrus urinary LH peak estradiol true peak time interval (h):

Figure CN103126728BD00081

[0047] 注:N表示样本数;a表示存在多个众数,只显示最小值。 [0047] Note: N represents the number of samples; A represents the presence of a plurality of the mode to display only the minimum.

[0048] 从表3可以看到,大熊猫LH峰值与雌二醇真实峰值间隔时间变异范围较大,从最小的O小时到最大的30.22小时,平均为13.67小时,其中大多数(60%)的个体在11.9小时到15.72小时即在约12~16小时之间。 [0048] can be seen from Table 3, the larger peak LH and estradiol panda true peak interval variation range, from the smallest to the largest O hours 30.22 hours, an average of 13.67 hours, the majority (60%) in individuals i.e. 11.9 hours to 15.72 hours between about 12 to 16 hours.

[0049] 研究结果表明,雌性大熊猫LH峰值时刻与雌激素峰值时刻间隔时间在不同个体间差异很大,在O~30.22小时之间,而LH峰值时刻到排卵的时间间隔通常是确定的。 [0049] The results show that female panda LH peak time and peak time interval estrogen great differences between different individuals, between O ~ 30.22 hours, ovulation LH peak time interval is typically determined. 研究过程还首次发现,相对于雌二醇真实峰值时刻,有部分大熊猫个体存在延迟排卵现象。 The study also found that for the first time, with respect to estradiol real peak time, there are some giant panda there is a delay ovulation phenomenon.

[0050] 排卵延迟理论上讲动物的发情高峰行为应随之相应延迟,为了证明排卵延迟是否会导致发情高峰行为延迟,对此做了进一步进行分析。 [0050] delay ovulation peak estrus animal behavior theory should be followed accordingly delay, delay ovulation in order to prove whether the peak estrus behavior will lead to delays, which made further analysis. 由于大熊猫通常只有在发情行为高峰时才接受本交、即自然交配,因此可将首次自然交配的时间作为发情行为高峰的标志,t匕较延迟排卵个体是否会导致首次本交时间的延迟。 Because pandas usually only accept the post only if the peak estrus behavior, namely natural mating, so you can be the first natural mating time as the peak estrus behavior logo, t dagger delay ovulation than whether an individual for the first time this will lead to a delay in delivery time. 以雌激素峰值时刻为零小时,并计算出LH峰值时间到雌激素峰值时间间隔,同时计算出发生自然交配的个体首次本交的时间,以LH到雌激素峰值间隔时间小于16小时个体为对照,比较LH到雌激素峰值间隔时间在16小时以上个体其本交时间是否会延迟,结果见下表4: Zero hour estrogen peak time, peak time and calculate the estrogen LH peak time interval, while the calculation time occurring natural mating individuals this first post, to the estrogen LH peak intervals less than 16 hours for the individual control Comparative LH estrogen to an individual peak intervals which present cross delay time is more than 16 hours will result in Table 4 below:

[0051] 表4.延迟排卵对首次本交时间的影响: [0051] Table 4. Effect of ovulation of the first delay time of the present post:

[0052] [0052]

Figure CN103126728BD00091

[0053] 从表4可以看到,延迟排卵组其本交时间也相应晚于对照组,由于大熊猫只有在发情行为高峰时才接受自然交配,因此上述数据证明延迟排卵个体其发情高峰行为相应延迟,自然交配时间从正常情况下的22.17小时延迟到距雌激素真实峰值后29.07小时,接近30小时。 [0053] can be seen from Table 4, the group delay ovulation which the present cross-time later than the corresponding control group, since the only accepted when estrus panda peak natural mating behavior, so the above data demonstrate that an individual delay ovulation oestrus behavior peak corresponding delay, the time delay from natural mating under normal circumstances 22.17 hours to 29.07 hours after the estrogen from the true peak, approaching 30 hours. 由于延迟排卵个体其本交时间相应延迟,因此对于延迟排卵个体应该实施延迟配种。 Since the delay time of ovulation subject which present respective cross-delayed, the delay should be an individual embodiment delay ovulation breeding.

[0054] 由于60%雌性大熊猫个体LH峰值到雌二醇真实峰值间隔时间在16小时以内,因此可以依据雌性大熊猫LH峰值时刻与雌二醇真实峰值时刻的时间间隔是否在16小时以内,初步判断雌性大熊猫是否正常或延迟排卵;同时伴随延迟排卵、其发情高峰行为相应延迟,可结合行为学指标进行综合判断。 [0054] Since the 60% peak-to-female giant panda LH estradiol true peak intervals less than 16 hours, can be based on the time female panda estradiol LH peak time interval is true peak time within 16 hours, female panda preliminary judgment whether normal or delay ovulation; accompanied by a delay ovulation, oestrus behavior peak corresponding delay can be combined with behavioral indicators comprehensive judgments. 由于延迟排卵个体自然交配均发生在大约30小时以后,再结合在雌二醇真实峰值时刻后的30小时内雌性大熊猫是否出现发情高峰行为,进一步确定雌性大熊猫是否正常或延迟排卵 Due to the delay ovulation individual natural mating occurred after about 30 hours, whether or not combined with female panda estrus peak behavior occurs within 30 hours after the peak estradiol real time, to further determine whether female panda normally or delay ovulation

[0055] 由于本发明以雌性大熊猫雌二醇真实峰值及其出现时刻为基础准确地检测出了雌性大熊猫LH峰值及其出现时刻,然后依据LH峰值时刻与雌二醇真实峰值时刻的时间间隔和雌性大熊猫是否出现发情高峰行为等条件共同确定雌性大熊猫正常或延迟排卵现象,准确地掌握了雌性大熊猫的排卵规律,为雌性大熊猫适时配种提供了适当时机,极大地促进了对濒危动物大熊猫的保护。 [0055] Since the present invention is estradiol in female panda true peak appears in time and accurately detected based on the peak of LH and female panda appearance time, peak time and then based on the time LH and estradiol true peak time female panda estrus interval and if a peak conditions jointly determine the behavior appears normal female panda or delay ovulation phenomenon, accurately grasp the law of ovulation female panda, providing timely appropriate time for the female panda breeding, which greatly promoted to protection of the endangered giant panda.

附图说明[0056] 下面结合附图和具体实施方式对本发明作进一步详细的说明。 BRIEF DESCRIPTION [0056] accompanying drawings and the following specific embodiments of the present invention will be further described in detail.

[0057]图1为本发明典型正常排卵雌性大熊猫个体的雌二醇值、孕酮值和LH值的变化曲线图; [0057] FIG 1 typical normal ovulatory estradiol female panda individual values ​​of the present invention, a graph showing changes in progesterone and LH values;

[0058] 图2为本发明典型延迟排卵雌性大熊猫个体的雌二醇值、孕酮值和LH值的变化曲线图。 [0058] FIG 2 is a typical delay ovulation estradiol female panda individual value change curve values ​​of LH and progesterone invention.

[0059] 图中标识:1_雌性大熊猫雌二醇值的变化曲线,2-雌性大熊猫孕酮值的变化曲线,3-雌性大熊猫LH值的变化曲线,A-雌二醇真实峰值坐标点,B-LH峰值坐标点,Ec ng/mg Cr表示为肌酐校正后的雌二醇浓度值,Pg ng/mg Cr表示为肌酐校正后的孕酮浓度值,LH mIU/mg Cr表示为肌酐校正后的LH浓度值。 [0059] FIG identified: 1_ female panda curve value estradiol, progesterone curve 2- female panda value curve of LH in female panda 3-, A- true peak estradiol coordinates of the point, B-LH peak coordinate point, Ec ng / mg Cr estradiol concentration values ​​expressed as creatinine correction, Pg ng / mg Cr concentration values ​​expressed as creatinine progesterone corrected, LH mIU / mg Cr expressed as LH concentration after creatinine correction.

具体实施方式 detailed description

[0060] 实施例一: [0060] Example a:

[0061] 本发明首先依照专利申请号201210191762.5所述,在春季繁育期间,按时间要求对雌性大熊猫进行尿样收集,对尿样采用酶联免疫吸附试验检测出雌性大熊猫雌二醇真实峰值及其出现时间,如图1所示,绘制出雌性大熊猫雌二醇值和孕酮值的变化曲线图,得到雌二醇真实峰值坐标点A。 [0061] First, in accordance with the present invention patent application No. 201210191762.5, breeding during the spring, time required for collecting urine female panda, urine sample using an enzyme-linked immunosorbent assay true peak estradiol female panda and the time of occurrence, as shown in FIG. 1, curves plotted in FIG female panda value estradiol and progesterone, and estradiol obtain true peak coordinate point A. 然后以坐标点A的时刻为时间起点O小时,使用上海拜沃生物科技有限公司的犬促黄体生成激素试剂盒,采用酶联免疫吸附试验检测O小时到48小时内全部尿样的LH值;将雌性大熊猫排尿时间作为横坐标,将LH值作为纵坐标,在图1上标出排尿时间及其对应的LH值所确定的坐标点,将坐标点依次连接起来,绘制出LH值变化曲线图,找出LH峰值及其出现时间,得到LH峰值坐标点B。 A coordinate point in time then the time of starting O hours, canine Haibai Wo luteinizing hormone kits Biotechnology Limited use of enzyme-linked immunosorbent assay value O LH hours all the urine within 48 hours; the female panda voiding time as the abscissa, the ordinate value as the LH, voiding time indicated on FIG. 1 and the coordinate point corresponding to the determined value of LH, in turn connects coordinate points plotted curve of LH FIG find the LH peak and the appearance time, peak LH obtained coordinate point B. 最后,计算出坐标点A与坐标点B的时间间隔为9.92小时,小于16小时,可初步确定雌性大熊猫为正常排卵个体;再根据坐标点A后的30小时内雌性大熊猫出现发情高峰行为,可进一步确定雌性大熊猫为正常排卵个体。 Finally, calculate the time point A and the coordinates of the coordinate point B spaced 9.92 hours, less than 16 hours may be initially identified as female panda individual normal ovulation; estrus behavior again according to the peak within 30 hours after the coordinate points A female panda , may further determine an individual female panda normal ovulation.

[0062] 实施例二: [0062] Example II:

[0063] 本发明首先依照专利申请号201210191762.5所述,在春季繁育期间,按时间要求对雌性大熊猫进行尿样收集,对尿样采用酶联免疫吸附试验检测出雌性大熊猫雌二醇真实峰值及其出现时间,如图2所示,绘制出雌性大熊猫雌二醇值和孕酮值的变化曲线图,得到雌二醇真实峰值坐标点A。 [0063] First, in accordance with the present invention patent application No. 201210191762.5, breeding during the spring, time required for collecting urine female panda, urine sample using an enzyme-linked immunosorbent assay true peak estradiol female panda and the time of occurrence, as shown in FIG. 2, the curve plotted values ​​in FIG female panda estradiol and progesterone, and estradiol obtain true peak coordinate point A. 然后以坐标点A的时刻为时间起点O小时,使用上海拜沃生物科技有限公司的犬促黄体生成激素试剂盒,采用酶联免疫吸附试验检测O小时到48小时内全部尿样的LH值;将雌性大熊猫排尿时间作为横坐标,将LH值作为纵坐标,在图2上标出排尿时间及其对应的LH值所确定的坐标点,将坐标点依次连接起来,绘制出LH值变化曲线图,找出LH峰值及其出现时间,得到LH峰值坐标点B。 A coordinate point in time then the time of starting O hours, canine Haibai Wo luteinizing hormone kits Biotechnology Limited use of enzyme-linked immunosorbent assay value O LH hours all the urine within 48 hours; the female panda voiding time as the abscissa, the ordinate value as the LH, voiding time indicated on FIG. 2 and the coordinate point corresponding to the determined value of LH, in turn connects coordinate points plotted curve of LH FIG find the LH peak and the appearance time, peak LH obtained coordinate point B. 最后,计算出坐标点A与坐标点B的时间间隔为23.17小时,大于16小时,可初步确定雌性大熊猫为延迟排卵个体;再根据坐标点A后的30小时内雌性大熊猫未出现发情高峰行为,可进一步确定雌性大熊猫为延迟排卵个体。 Finally, to calculate the coordinate points of time A and B coordinate point interval of 23.17 hours, more than 16 hours, initially identified as a female panda individual delay ovulation; estrus peak does not appear after 30 hours according to the coordinates of the point A female panda behavior, can further determine an individual female panda delay ovulation.

[0064]本发明并不局限于以上实例,本发明的LH峰值及其出现时间不一定通过直观的变化曲线图找出,可以只通过比较所有检测出的LH值大小得到。 [0064] The present invention is not limited to the above examples, the present invention LH peak and the time of occurrence of the change is not necessarily intuitive graph to find, can only compare all detected LH values ​​obtained by size.

Claims (4)

  1. 1.一种大熊猫正常或延迟排卵的确定方法,包括春季繁育期间,按时间要求对雌性大熊猫进行尿样收集,然后对尿样采用酶联免疫吸附试验检测出雌性大熊猫雌二醇真实峰值及其出现时间的步骤,其特征在于还包括以下步骤: (1)以出现雌性大熊猫雌二醇真实峰值的时刻为时间起点O小时,采用酶联免疫吸附试验检测O小时到48小时内全部尿样的促黄体生成激素值,找出促黄体生成激素峰值及其出现时间; (2)依据雌二醇真实峰值时刻、促黄体生成激素峰值时刻与雌二醇真实峰值时刻的时间间隔、雌性大熊猫是否出现发情高峰行为等条件共同确定雌性大熊猫正常或延迟排卵现象。 1. A method of determining a normal or delay ovulation pandas, including breeding during the spring, time required for collecting urine female panda, and a urine sample by enzyme linked immunosorbent assay detected authentic estradiol female panda its peak appearance time step, characterized by further comprising the steps of: (1) occurs in real time, a female panda estradiol peak time of starting O hours, with the test for detection of O to 48 hours ELISA all urine samples of luteinizing hormone values, find luteinizing hormone peak and time of occurrence; (2) based on estradiol real peak time, time luteinizing hormone and estradiol real peak time peak time interval, female panda estrus peak whether the conditions jointly determine the behavior appears normal female panda or delay ovulation phenomena.
  2. 2.根据权利要求1所述的一种大熊猫正常或延迟排卵的确定方法,其特征在于:当雌性大熊猫促黄体生成激素峰值时刻与雌二醇真实峰值时刻的时间间隔小于16小时、且在雌二醇真实峰值时刻后的30小时内雌性大熊猫出现发情高峰行为时,则可以确定雌性大熊猫为正常排卵个体。 2. According to a panda according to claim 1 or method of determining the normal delay ovulation, characterized in that: when a female panda time luteinizing hormone and estradiol true peak peak time interval is less than 16 hours of time, and when female panda estrus peak behavior occurs within 30 hours after the estradiol real peak time, it can be determined to be normal ovulation female panda individuals.
  3. 3.根据权利要求1所述的一种大熊猫正常或延迟排卵的确定方法,其特征在于:当雌性大熊猫促黄体生成激素峰值时刻与雌二醇真实峰值时刻的时间间隔大于16小时、且在雌二醇真实峰值时刻后的30小时内雌性大熊猫未出现发情高峰行为时,则可以确定雌性大熊猫为延迟排卵个体。 3. According to a panda according to claim 1 or method of determining the normal delay ovulation, characterized in that: when a female panda time luteinizing hormone and estradiol true peak peak time interval is greater than 16 hours of time, and when female panda estrus peak behavior not appear within 30 hours after the estradiol real peak time, you can determine the individual female panda delay ovulation.
  4. 4.根据权利要求1~3所述的任何一种大熊猫正常或延迟排卵的确定方法,其特征在于:促黄体生成激素值采用犬促黄体生成激素试剂盒检测。 The panda any one of the claims 1 to 3, or the method for determining the normal delay ovulation, characterized in that: the value of luteinizing hormone in dogs using luteinizing hormone kit.
CN 201310070672 2013-03-06 2013-03-06 Determination method of normal or delayed ovulation in pandas CN103126728B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN 201310070672 CN103126728B (en) 2013-03-06 2013-03-06 Determination method of normal or delayed ovulation in pandas

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN 201310070672 CN103126728B (en) 2013-03-06 2013-03-06 Determination method of normal or delayed ovulation in pandas

Publications (2)

Publication Number Publication Date
CN103126728A true CN103126728A (en) 2013-06-05
CN103126728B true CN103126728B (en) 2014-03-19

Family

ID=48487626

Family Applications (1)

Application Number Title Priority Date Filing Date
CN 201310070672 CN103126728B (en) 2013-03-06 2013-03-06 Determination method of normal or delayed ovulation in pandas

Country Status (1)

Country Link
CN (1) CN103126728B (en)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1129898A (en) 1993-07-02 1996-08-28 尤尼帕思有限公司 Method, apparatus and testing box for determining ovulation
CN101528138A (en) 2006-09-05 2009-09-09 孕育中心有限公司 Method of detecting and predicting ovulation and the period of fertility
WO2010061613A1 (en) 2008-11-28 2010-06-03 株式会社日立製作所 Method, system and kit for supporting pregnancy
CN102735852A (en) 2012-06-12 2012-10-17 成都大熊猫繁育研究基地 Determination method of giant panda estradiol true peak value
CN102743239A (en) 2012-07-02 2012-10-24 成都大熊猫繁育研究基地 Timely artificial insemination method for giant panda

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1129898A (en) 1993-07-02 1996-08-28 尤尼帕思有限公司 Method, apparatus and testing box for determining ovulation
CN101528138A (en) 2006-09-05 2009-09-09 孕育中心有限公司 Method of detecting and predicting ovulation and the period of fertility
WO2010061613A1 (en) 2008-11-28 2010-06-03 株式会社日立製作所 Method, system and kit for supporting pregnancy
CN102735852A (en) 2012-06-12 2012-10-17 成都大熊猫繁育研究基地 Determination method of giant panda estradiol true peak value
CN102743239A (en) 2012-07-02 2012-10-24 成都大熊猫繁育研究基地 Timely artificial insemination method for giant panda

Also Published As

Publication number Publication date Type
CN103126728A (en) 2013-06-05 application

Similar Documents

Publication Publication Date Title
Odell et al. Radioimmunoassay for human follicle—stimulating hormone: physiological studies
AIMAN et al. The frequency of androgen receptor deficiency in infertile men
Smith et al. Serum levels of luteinizing hormone and progesterone during the estrous cycle, pseudopregnancy and pregnancy in the dog
Garth Sasser et al. Detection of pregnancy by radioimmunoassay of a novel pregnancy-specific protein in serum of cows and a profile of serum concentrations during gestation
Savabieasfahani et al. Developmental programming: differential effects of prenatal exposure to bisphenol-A or methoxychlor on reproductive function
Radford et al. Ovarian function in suckling and non-suckling beef cows post partum
Wilcox et al. Post-ovulatory ageing of the human oocyte and embryo failure
Nuojua-Huttunen et al. Intrauterine insemination treatment in subfertility: an analysis of factors affecting outcome
Bell et al. Chronic heat stress and prenatal development in sheep: I. Conceptus growth and maternal plasma hormones and metabolites
Burger et al. Hormonal changes in the menopause transition
Gonzalez-Bulnes et al. Effects of progestagens and prostaglandin analogues on ovarian function and embryo viability in sheep
Schwarzenberger et al. Concentrations of faecal immunoreactive progestagen metabolites during the oestrous cycle and pregnancy in the black rhinoceros (Diceros bicornis michaeli)
Shih et al. Factors affecting low birthweight after assisted reproduction technology: difference between transfer of fresh and cryopreserved embryos suggests an adverse effect of oocyte collection
Gould et al. Chimpanzee reproductive senescence: a possible model for evolution of the menopause
Brown et al. Hormone Secretion in the Asian Elephant (Elephas maximus): Characterizationof Ovulatory and Anovulatory Luteinizing Hormone Surges
Moreira et al. Reproductive steroid hormones and ovarian activity in felids of the Leopardus genus
Shideler et al. Monitoring ovulation and implantation in the lion-tailed macaque (Macaca silenus) through urinary estrone conjugate evaluations
Ziegler et al. Urinary gonadotropin and estrogen excretion during the postpartum estrus, conception, and pregnancy in the cotton‐top tamarin (Saguinus oedipus oedipus)
Out et al. A prospective, randomized, assessor-blind, multicentre study comparing recombinant and urinary follicle stimulating hormone (Puregon versus Metrodin) in in-vitro fertilization
Lefevre et al. Reproductive senescence in female rats: a longitudinal study of individual differences in estrous cycles and behavior
Ehrlich et al. Urinary bisphenol A concentrations and implantation failure among women undergoing in vitro fertilization
Burke et al. Estradiol benzoate delays new follicular wave emergence in a dose-dependent manner after ablation of the dominant ovarian follicle in cattle
Walker et al. Urinary concentrations of ovarian steroid hormone metabolites and bioactive follicle-stimulating hormone in killer whales (Orcinus orchus) during ovarian cycles and pregnancy
Rutigliano et al. Effects of method of presynchronization and source of selenium on uterine health and reproduction in dairy cows
Martin et al. Effects of dietary zinc deficiency on the reproductive system of young male sheep: testicular growth and the secretion of inhibin and testosterone

Legal Events

Date Code Title Description
C06 Publication
C10 Request of examination as to substance