Krueger C, Rath D, Johnson L A
Institute of Animal Science and Animal Behaviour, Mariensee, Neustadt, Germany.
Theriogenology. 1999 Dec;52(8):1363-73. doi: 10.1016/s0093-691x(99)00223-x.
Conventional insemination techniques in pigs require 2 to 3 x 10(9) sperm/dose. When using the latest high-speed sperm-sorting technology, one can still sort only about 5 to 6 million sperm of each sex per hour. The objective of the present study was to find the minimal sperm concentration at a low-insemination volume in pigs without diminishing fertilization rate and litter size using surgical deep intra-uterine insemination (IUI). Semen from 3 boars was collected and diluted with Androhep to 5 x 10(8), 1 x 10(8), 1 x 10(7), 5 x 10(6) or 1 x 10(6) sperm/0.5 ml. In trial 1, 109 prepuberal gilts were synchronized and surgically inseminated into the tip of each uterine horn 32 h or 38 h after hCG treatment or at the time of ovulation, respectively. Pregnant gilts were allowed to go to term. Pregnancy and farrowing rates did not differ significantly except at the lowest sperm concentration if inseminated 32 h or 38 h after hCG treatment (p < 0.05). No differences were found among insemination groups for the total number of piglets, number of piglets born alive, stillborn piglets, and mummified fetuses. In trial 2, 34 gilts were inseminated as described above 32 h after hCG. Additionally, 9 gilts were inseminated once nonsurgically with 1 x 10(9) sperm as controls. Gilts were slaughtered 48 h after insemination, and embryos were recovered. Embryos were cultured in NCSU 23 (120 h), evaluated morphologically and stained with fluorescent dye (Hoechst 33342) to visualize nuclei. Recovery rates varied between 71.4% and 84.4%. Fertilization rate of the lowest sperm concentration (1 x 10(6) sperm/horn) differed significantly (p < 0.05) from all other groups. Cleavage rates at specific developmental stages did not differ. After 5 days of in vitro culture, embryos developed to morulae and blastocysts. No differences were found for these stages. In conclusion, no major differences were found between insemination groups as long as the sperm dosage was at least 10 million sperm per gilt. The low volume was sufficient for successful deep intra-uterine insemination. Embryo development was comparable to the controls.
猪的传统授精技术每剂量需要2至3×10⁹个精子。使用最新的高速精子分选技术时,每小时仍只能分选大约500万至600万个不同性别的精子。本研究的目的是通过手术深部子宫内授精(IUI),找出猪在低授精量情况下不降低受精率和产仔数的最低精子浓度。采集3头公猪的精液,用Androhep稀释至5×10⁸、1×10⁸、1×10⁷、5×10⁶或1×10⁶个精子/0.5毫升。在试验1中,109头青春期前的后备母猪进行同期发情处理,分别在hCG处理后32小时或38小时或排卵时通过手术将精液授入每个子宫角的顶端。怀孕的后备母猪使其足月分娩。除了在hCG处理后32小时或38小时授精时最低精子浓度组外,妊娠率和产仔率没有显著差异(p<0.05)。在仔猪总数、活产仔猪数、死产仔猪数和木乃伊化胎儿数方面,各授精组之间没有差异。在试验2中,34头后备母猪在hCG处理后32小时按上述方法授精。另外,9头后备母猪作为对照非手术授精1×10⁹个精子一次。授精后48小时屠宰后备母猪并回收胚胎。胚胎在NCSU 23中培养(120小时),进行形态学评估并用荧光染料(Hoechst 33342)染色以观察细胞核。回收率在71.4%至84.4%之间。最低精子浓度组(1×10⁶个精子/子宫角)的受精率与所有其他组有显著差异(p<0.05)。特定发育阶段的分裂率没有差异。体外培养5天后,胚胎发育至桑葚胚和囊胚阶段。这些阶段没有差异。总之,只要每头后备母猪的精子剂量至少为1000万个精子,各授精组之间没有重大差异。低体积足以成功进行深部子宫内授精。胚胎发育与对照组相当。