CN107333704A - 提高母畜体内早期胚胎附植数量和窝产仔数的方法 - Google Patents
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Abstract
本发明涉及动物繁殖领域，具体公开了提高母畜体内早期胚胎附植数量和窝产仔数的方法以及N‑乙酰半胱氨酸(NAC)在提高母畜体内早期胚胎附植数量和窝产仔数方面的应用。本发明以小鼠、猪为例，通过摄入NAC，针对性地改善了子宫环境及提高早期胚胎发育能力，增加附植胚胎数量，具有提高母畜窝产仔数的能力；且NAC属小分子化合物，化学性质较稳定，生产成本较低，具有广阔的应用前景和巨大的经济效益。
Description
技术领域
本发明涉及动物繁殖领域，具体地说，涉及提高母畜体内早期胚胎附植数量和窝产仔数的方法。
背景技术
超数排卵(Superovulation，简称超排)，是应用外源性促性腺激素诱发卵巢多个卵泡发育，排出比自然条件下多几倍甚至十几倍并具有受精能力的卵子，目的是在优良母畜有效繁殖年限内，尽可能获得其后代，快速扩繁优良种群，或者获得较多的卵子用于体外受精和早期胚胎的培养。目前，超数排卵主要采用血清促性腺激素(PMSG)刺激卵泡发育，再经人绒毛膜促性腺激素释放激素(hCG)或促黄体生成素(LH)以及促性腺激素释放激素类似物(GnRH-a)诱导排卵，以获得较多的卵子。尽管超排技术在生产实践及实验室应用相当广泛，但超排会对母体子宫或胚胎造成伤害，如卵泡异质性增加(ManyMC,Denef JF,Hamudi S,Haumont S.Increased follicular heterogeneityin experimental colloid goiter produced by refeeding iodine excess afterthyroid hyperplasia.Endocrinology 1986；118:637-644.)，排卵虽然增加但胚胎活力降低(Do LV,Vianna W,Gama R,Campos RA,Pinese M,Sant'Anna MA.Second oestrus synchronization and precocious embryoviability after puberty induction in gilts by the use of gonadotrophintreatment.Reprod Domest Anim 2005；40:141-144.Martinat-Botte F,Venturi E,Guillouet P,Driancourt MA,Terqui M.Induction andsynchronization of ovulations of nulliparous and multiparous sows withan injection of gonadotropin-releasing hormone agonist(Receptal).Theriogenology 2010；73:332-342.)，或排卵增加但附植的胚胎并未显著增加(Martinat-Botte F,Venturi E,Guillouet P,Driancourt MA,TerquiM.Induction and synchronization of ovulations of nulliparous andmultiparous sows with an injection of gonadotropin-releasing hormoneagonist(Receptal).Theriogenology 2010；73:332-342.)，最终产仔数亦未提高(Longenecker DE,Day BN.Fertility level of sows superovulatedat post-weaning estrus.J Anim Sci 1968；27:709-711)。研究结果显示应用PMSG进行超数排卵或者同期发情改变了子宫内分泌环境(PopeWF,Xie S,Broermann DM,Nephew KP.Causes and consequences ofearly embryonic diversity in pigs.J Reprod Fertil Suppl 1990；40:251-260.)，使子宫环境与早期胚胎发育不同步从而影响了早期胚胎发育。
近年来，规模化、集约化畜禽养殖快速发展，提高多胎动物窝产仔数已经成为养殖业增收的重要途径。自1949年，Tanabe等首次用PMSG刺激猪卵泡发育后，PMSG已经被广泛应用于实验室小鼠超排取卵，或牛、羊超排冲胚等实践生产过程。但文献研究表明，超数排卵后成功附植的胚胎以及产仔数并没有随之增加。因此，针对这一问题，研究人员尝试通过向超排母猪日粮添加一定比例的叶酸，但并未提高经产母猪产仔数(Longenecker DE,Day BN.Fertility level of sowssuperovulated at post-weaning estrus.J Anim Sci 1968；27:709-711.)；虽然文献表明对未经PMSG超排处理的雌性CD-1小鼠通过腹腔注射褪黑素会提高产仔数(He C,Wang J,Li Y,Zhu K,Xu Z,Song Y,Song Y,Liu G.Melatonin-related genes expressed in the mouse uterus duringearly gestation promote embryo implantation.J Pineal Res 2015；58:300-309.)，但长期使用褪黑素会影响自身内分泌，甚至产生依赖性。且由于褪黑素见光易分解，所有操作需避光进行，导致处理过程繁琐；日粮添加N-氨甲酰谷氨酸(NCG)可以提高大鼠及母猪产仔数(ZhuJ,Zeng X,Peng Q,Zeng S,Zhao H,Shen H,Qiao S.MaternalN-Carbamylglutamate Supplementation during Early PregnancyEnhances Embryonic Survival and Development through Modulation ofthe Endometrial Proteome in Gilts.J Nutr 2015；145:2212-2220.)，但其是否能提高超排母畜产仔数，暂时没有相关研究；在利用PMSG对母畜进行同期发情处理或定时输精的时代背景及提高养殖企业和农户经济效益的迫切需求下，增加母畜的产仔数是一种必然趋势。目前，针对超排后早期胚胎附植能力低，产仔数未增加的研究较少，目前暂未取得令人满意的成果。
发明内容
为了解决现有技术中存在的问题，本发明的目的是提供一种提高母畜体内早期胚胎附植数量和窝产仔数的方法。
为了实现本发明目的，本发明的技术方案如下：
本发明针对PMSG可以提高雌性排卵，但早期胚胎附植或最终产仔数并未提高的技术问题，以小鼠为模式动物，通过腹腔注射N-乙酰半胱氨酸(NAC)改善母畜子宫环境、提高小鼠早期胚胎附植能力，以期最终提高母畜窝产仔数。
本发明基于上述研究，科学合理地确定了NAC注射剂量及时间，有效提高了雌性小鼠体内早期胚胎附植数量，并可提高母畜最终窝产仔数。
第一方面，本发明提供了N-乙酰半胱氨酸在提高母畜体内早期胚胎附植数量和窝产仔数方面的应用。
第一方面，本发明提供了一种提高母畜体内早期胚胎附植数量的方法，使体成熟的动物通过注射或饲喂的方式摄取N-乙酰半胱氨酸。
进一步地，通过注射方式摄取N-乙酰半胱氨酸的剂量为50～300mg/kg BW·天，通过饲喂方式摄取N-乙酰半胱氨酸的剂量为1～300mg/kg BW·天。
更进一步地，当所述动物为小鼠时，通过注射方式摄取N-乙酰半胱氨酸的剂量为100～200mg/kg BW·天，优选150mg/kgBW·天。
更进一步地，当所述动物为猪时，通过饲喂方式摄取N-乙酰半胱氨酸的剂量为5～10mg/kg BW·天，优选7.5mg/kg BW·天。
更为具体地，在所述动物摄取N-乙酰半胱氨酸的期间，对其进行超数排卵处理，并保证其摄取N-乙酰半胱氨酸直至胚胎附植结束。
进一步地，本发明在具体实施方式中以小鼠和猪为例进行试验，但本发明所述动物为多胎动物，并不限于猪和小鼠。
第三方面，本发明提供了一种提高母畜窝产仔数的方法，具体为：使多胎动物自交配处理前至少7天起通过注射或饲喂的方式摄取N-乙酰半胱氨酸，直至胚胎附植或分娩结束。
本发明的有益效果在于：
本发明通过腹腔注射NAC，针对性地改善了PMSG处理后的子宫环境及提高小鼠体内早期胚胎发育能力，增加了超排小鼠附植胚胎数量，具有提高母畜窝产仔数的能力；且NAC属小分子化合物，化学性质较稳定，生产成本较低，具有广阔的应用前景和巨大的经济效益。
附图说明
图1为本发明实施例1中实验组的操作简图。
具体实施方式
下面将结合实施例对本发明的优选实施方式进行详细说明。需要理解的是以下实施例的给出仅是为了起到说明的目的，并不是用于对本发明的范围进行限制。本领域的技术人员在不背离本发明的宗旨和精神的情况下，可以对本发明进行各种修改和替换。
下述实施例中所使用的实验方法如无特殊说明，均为常规方法。
下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
实施例1提高小鼠体内早期胚胎附植能力
1、实验组
随机选取7～8周龄ICR雌性小鼠(北京维通利华)，如图1所示，每天17:00通过腹腔注射0.1mL NAC溶液(Sigma公司)，NAC注射量相当于150mg/kg体重，连续注射14天，并在第5天17:00注射NAC后，接着注射5IU PMSG(宁波第二激素厂)，48h后(注射NAC的第7天)注射5IU hCG(宁波第二激素厂)，并将雌性小鼠与雄性ICR小鼠按1:1同笼交配，次日早8:00检栓，见栓当天记为0.5dpc(第8天)。在7.5dpc(第15天)时，通过尾静脉向妊娠小鼠注射浓度为1％的芝加哥蓝，5min后脱颈处死，用灭菌剪刀及手术镊子打开腹腔，小心剥离子宫，计数胚胎附植位点。
2、对照组
将NAC注射液换成同体积的生理盐水，其他操作同实验组。
3、实验结果
表1腹腔注射NAC对小鼠早期胚胎附植能力的影响
注：同列数据肩标字母不同表示差异显著(P<0.05)
实施例2NAC提高母猪窝产仔数
将体重无明显差异的妊娠母猪分为两组，设置对照组(n＝19)和试验组(n＝26)。
对照组的日粮中未添加NAC，试验组的日粮中500mg/kg的NAC。
两组母猪自产前7天开始饲喂上述不同日粮，直至分娩结束，统计窝产仔数并对比，见表2。
表2日粮中添加NAC对母猪窝产仔数的影响
注：同行数据肩标不同表示差异显著(P<0.05)
实验结果表明，通过饲喂方式摄取NAC的母猪相对于未摄取NAC的母猪，可以显著提高窝产仔数。
虽然，上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述，但在本发明基础上，可以对之作一些修改或改进，这对本领域技术人员而言是显而易见的。因此，在不偏离本发明精神的基础上所做的这些修改或改进，均属于本发明要求保护的范围。
Claims (10)
1.N-乙酰半胱氨酸在提高母畜体内早期胚胎附植数量和窝产仔数方面的应用。
2.一种提高母畜体内早期胚胎附植数量的方法，其特征在于，使体成熟的动物通过注射或饲喂的方式摄取N-乙酰半胱氨酸。
3.根据权利要求2所述的方法，其特征在于，通过注射方式摄取N-乙酰半胱氨酸的剂量为50～300mg/kg BW·天，通过饲喂方式摄取N-乙酰半胱氨酸的剂量为1～300mg/kg BW·天。
4.根据权利要求3所述的方法，其特征在于，所述动物为小鼠，通过注射方式摄取N-乙酰半胱氨酸的剂量为100～200mg/kg BW·天。
5.根据权利要求4所述的方法，其特征在于，通过注射方式摄取N-乙酰半胱氨酸的剂量为150mg/kg BW·天。
6.根据权利要求3所述的方法，其特征在于，所述动物为猪，通过饲喂方式摄取N-乙酰半胱氨酸的剂量为5～10mg/kg BW·天。
7.根据权利要求6所述的方法，其特征在于，通过饲喂方式摄取N-乙酰半胱氨酸的剂量为7.5mg/kg BW·天。
8.根据权利要求1～7任一项所述的方法，其特征在于，在所述动物摄取N-乙酰半胱氨酸的期间，对其进行超数排卵处理，并保证其摄取N-乙酰半胱氨酸直至胚胎附植或分娩结束。
9.根据权利要求8所述的方法，其特征在于，所述动物为多胎动物。
10.一种提高母畜窝产仔数的方法，其特征在于，使多胎动物自交配处理前至少7天起通过注射或饲喂的方式摄取N-乙酰半胱氨酸，直至胚胎附植或分娩结束。
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