CN117298256A - 一种提高牦牛生殖性能的促性腺激素组合物及其施用方法 - Google Patents
一种提高牦牛生殖性能的促性腺激素组合物及其施用方法 Download PDFInfo
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Abstract
一种提高牦牛生殖性能的促性腺激素组合物及其施用方法,该促性腺激素组合物由重组马促卵泡生成素(reFSH)和重组人绒毛膜促性腺激素(rhCG)组成,以每头份单次注射量计算,其包含1500‑2500IU的reFSH和300‑600IU的rhCG。本发明的组合物特别用于治疗或恢复乏情牦牛的发情和诱导产犊牦牛的同期发情,在注射本发明组合物后,乏情牦牛恢复发情或同期发情,发情周期显著缩短,且受配率和受胎率均不受影响。
Description
说明书
相关申请
本申请要求申请日为2022年6月22日提交的中国专利申请号 CN202210714709.2的优先权,其全部内容通过引用并纳入本文。
技术领域
本发明涉及一种提高牦牛生殖性能的促性腺激素组合物,具体而言,涉及一种能诱导牦牛发情及同期发情的促性腺激素组合物和施用方法。
发明背景
牦牛(Bos grunniens)是高原地区的特殊畜种,也是唯一能适应特殊寒冷气候条件的牛种,但高原严寒、缺氧、缺草等恶劣环境通常造成牦牛的繁殖性能低下,呈现2年1胎或3年1胎的繁殖模式。生殖激素药物诱导牦牛发情或同期发情是提高牦牛繁殖性能的重要手段,这些激素包括雌激素、孕激素、促性腺激素释放激素(GnRH)、促卵泡激素(FSH)和促黄体激素(LH)等。一直以来,生殖激素调控“下丘脑-垂体-卵巢”轴诱导牦牛发情或同步发情是最为有效的手段,如以孕酮控释(CIDR)联合氯前列烯醇(PG)等可诱导牦牛发情或同期发情(梁春年等,中国专利CN103784212A);以雌激素、黄体酮、睾酮组合的三合激素处理可通过调节卵巢诱导母牦牛发情及同期发情,虽然诱导的发情率高但受胎率很低。陈世彪等以GnRH联合PGF2a的处理方案可诱导母牦牛同期发情,虽然发情率低于三合激素,但受胎率显著增加(“畜牧与兽医”,2014)。王文飙等(“中国牛业科学”,2017)采用氯前列烯醇(PG)联合GnRH或联合孕马血清促性腺激素(PMSG)处理诱导的牦牛同期发情率分别是91%和94%,妊娠率分别是 73%和69%。目前认为能调控牦牛发情或同期发情,以及提高受胎率的生殖激素主要是GnRH和PMSG,但二者单独应用十分有限,通常是二者合用或与其他激素联合应用,但实施方案的周期较长。
发明内容
本发明的首要目的是为改善牦牛繁殖性能低下而提供一种诱导乏情牦牛恢复发情和产犊牦牛同期发情的促性腺激素组合物。
更具体地说,本发明直接从调控卵泡生长和成熟的同步化方面,提供了重组 FSH和hCG的组合物用于提高牦牛的生殖性能。马源FSH和牛源FSH高度同源,本发明人在体外研究中发现,特异表达牛FSH受体(eFSHR)的细胞对重组马 FSH(reFSH)的反应优于重组牛FSH(rbFSH),在促进eFSHR特异表达细胞的 cAMP合成、原代牛卵巢颗粒细胞的雌激素和孕激素合成方面尤其显著。因此,在该组合物中,reFSH替代牛FSH的作用,rhCG替代牛LH的作用。在本发明的组合物中添加了rhCG,可协同reFSH促进颗粒细胞和黄体细胞合成雌激素和孕激素,也有利于腔前卵泡发育形成卵泡腔,并促进卵泡的最终成熟。
优选地,对乏情母牦牛实施单次交巢穴注射或肌肉注射本发明组合物,每头份施用生物活性剂量为1500-2500IU的reFSH和300-600IU的rhCG,注射次日后连续观察发情体征,待母牦牛发情后或配种前适时注射1000IU的rhCG促进排卵,随后实施与公牦牛配种或人工输精。
更优选地,对乏情母牦牛实施单次交巢穴注射或肌肉注射本发明组合物,每头份施用生物活性剂量为2000IU(reFSH)+500IU(rhCG);注射次日后连续观察发情体征,待母牦牛发情后或配种前适时注射1000IU的rhCG促进排卵,随后实施与公牦牛配种或人工输精。
更优选地,乏情母牦牛实施2次交巢穴注射或肌肉注射本发明组合物,首次每头份施用生物活性剂量为1800IU(reFSH)+450IU(rhCG),次日每头份再次注射 1800IU(reFSH)+450IU(rhCG)。于第2次注射后连续观察发情体征,待发情后或配种前适时注射1000IU的rhCG促进排卵,随后实施与公牦牛配种或人工输精。
本发明的另一目的是提供诱导带犊母牦牛同期发情的促性腺激素组合物的施用方法,操作简便、耗时短且十分有效。
优选地,对带犊母牦牛实施单次交巢穴注射或肌肉注射本发明组合物,每头份施用生物活性剂量为1800IU的reFSH和450IU的rhCG,注射次日后连续观察发情体征,待母牦牛发情后或配种前适时注射1000IU的rhCG促进排卵,随后实施与公牦牛配种或人工输精。
更优选地,对带犊母牦牛单次交巢穴注射或肌肉注射本发明组合物,每头份施用生物活性剂量为2000IU的reFSH和500IU的rhCG,注射次日后连续观察发情体征,待母牦牛发情后或配种前适时注射1000IU的rhCG促进排卵,随后实施与公牦牛配种或人工输精。
本发明的有益之处:操作简单、用时短且效果十分明显,为改善高原牦牛繁殖性能低下、牦牛隔年甚至3年一胎的繁殖模式,提高和发展牦牛产业具有重要意义。
下面结合实施例对本发明作进一步说明。
实施例1本发明组合物的配制
组合物1的配制:无菌条件下,取注射用reFSH与注射用rhCG混匀,加入生理盐水或注射用水至所需体积,随后以5mL量分装至西林瓶,每瓶含reFSH (2000IU)和rhCG(500IU)。
组合物2的配制:无菌条件下,取注射用reFSH与注射用rhCG混匀,加入生理盐水或注射用水至所需体积,随后以5mL量分装至西林瓶,每瓶含reFSH (1800IU)和rhCG(450IU)。
实施例2单次注射本发明对乏情母牦牛恢复发情的影响
选择隔年或两年未自然发情的母牦牛256头(体重200-250公斤),在发情季节施用本发明。将所选择的乏情母牦牛集中饲养管理,于1周后单次交巢穴注射本发明组合物1(下称实验组1),以每头份注射5mL,其包含2000IU的reFSH 和500IU的rhCG,注射次日后连续观察发情体征,待母牦牛发情后适时注射 1000IU的rhCG,每头份注射量为5mL,随后实施与公牦牛配种。另选择120头自然放牧和不经任何注射处理的乏情母牛,作为本发明组合物1的对照组。从起始发情的第1只母牦牛开始观察和计算,结果显示,256头乏情母牦牛经本发明组合物1处理后,224头乏情牦牛恢复发情,发情率为87.5%(表1),同期妊娠的受配率为93%,次年顺利产犊(受胎率为100%)。同比对照,120头自然放牧的对照组牦牛在发情季节仅有10头出现发情体征,发情率仅为8.33%。此外,对照组牦牛发情持续时长为7-21天,本发明组合物1处理后母牦牛的发情持续时长显著缩短为3-6天。
表1单次注射本发明对乏情母牦牛恢复发情的影响
实施例3两次注射本发明对乏情母牦牛恢复发情的影响
选择隔年或两年未自然发情的母牦牛32头(体重200-250公斤),在发情季节施用本发明。将所选择的乏情母牦牛集中饲养管理,于1周后首次交巢穴注射本发明组合物2(以下称实验组2),以每头份注射5mL,其包含1800IU的reFSH 和450IU的rhCG;于第2日以相同量再次注射本发明组合物2,注射次日连续观察发情体征,待母牦牛发情后实施与公牦牛配种,同步交巢穴注射1000IU的 rhCG,每头份注射量为5mL。另选择16头自然放牧和不经任何注射处理的乏情母牛,作为本发明组合物2的对照组。
从两次注射后起始发情的第1只母牦牛开始观察和计算,结果显示,32头乏情母牦牛经本发明组合物2处理后,31头恢复发情,发情率为96.88%(表2),发情持续时长为3-5天。同比对照,16头自然放牧的对照组牦牛在发情季节仅1 头在第19天出现发情体征(发情率仅为6.25%)。
表2两次注射本发明对乏情母牦牛恢复发情的影响
实施例4单次注射本发明诱导断奶母牦牛的同期发情和定时输精
选择带犊母牦牛273头(体重150-200公斤),均于8月初的同一天断乳,断乳后交巢穴注射本发明组合物1(以下称实验组3),以每头份注射5mL,其包含2000IU的reFSH和500IU的rhCG,注射次日后连续观察发情体征,从起始发情的第1只断乳母牦牛开始观察和计算,结果显示,注射本发明组合物1后,断乳母牦牛均在52小时左右出现发情,发情时间集中且发情率达到100%(见表3)。于发情后适时注射1000IU的rhCG,每头份注射量为5mL,同时实施定时输精。同比对照,未经本发明处理的231头断乳母牦牛中有148头于断奶后自然发情,发情率为64.07%,发情时长为7-15天。
表3单次注射本发明对断奶母牦牛同期发情的影响
Claims (5)
1.一种提高牦牛生殖性能的促性腺激素组合物,由重组马促卵泡生成素(reFSH)和重组人绒毛膜促性腺激素(rhCG)组成,其特征在于,以每头份计算,reFSH的生物活性单位为1500-2500IU,rhCG的生物活性单位为300-600IU。
2.如权利要求1所述的促性腺激素组合物,其特征在于,以每头份计算,reFSH的生物活性单位为2000IU,rhCG的生物活性单位为500IU。
3.权利要求1和2所述促性腺激素组合物在诱导乏情母牦牛发情的施用方法,其特征在于,对乏情母牦牛实施单次交巢穴注射或肌肉注射促性腺激素组合物,每头份施用生物活性剂量为1500-2500IU的reFSH和300-600IU的rhCG,优选生物活性剂量为2000IU(reFSH)+500IU(rhCG);注射次日后连续观察发情体征,待母牦牛发情后或配种前适时注射1000IU的rhCG,随后实施与公牦牛配种或人工输精。
4.权利要求1和2所述促性腺激素组合物在诱导乏情母牦牛发情的施用方法,其特征在于,对乏情母牦牛实施2次交巢穴注射或肌肉注射促性腺激素组合物,首次每头份施用生物活性剂量为1800IU(reFSH)+450IU(rhCG),次日每头份再次交巢穴注射或肌肉注射1800IU(reFSH)+450IU(rhCG)。于第2次注射后连续观察发情体征,待发情后或配种前适时注射1000IU的rhCG,随后实施与公牦牛配种或人工输精。
5.权利要求1和2所述促性腺激素组合物在诱导带犊母牦牛同期发情的施用方法,其特征在于,对带犊母牦牛在断奶后实施单次交巢穴注射或肌肉注射促性腺激素组合物,每头份施用生物活性剂量为1500-2500IU的reFSH和300-600IU的rhCG,优选生物活性剂量为2000IU(reFSH)+500IU(rhCG);注射次日后连续观察发情体征,待发情后或配种前适时注射1000IU的rhCG,随后同步实施人工输精或定时输精。
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