It is known that sperm cells will settle out of extender over time which could impact the concentration per dose within a batch. The objective of this study to investigate the effects of mixing prior to packaging on concentration of packaged insemination doses. At a commercial boar stud, three ejaculates were collected, extended, pooled and assigned to one of three treatment groups: pour-mix (PM) where ejaculates were immediately packaged after combining the three ejaculates, stir-mix (SM) where three ejaculates were combined and stirred thoroughly then packaged, or no mixing (NM) where pooled ejaculates were allowed to sit for 10 min then packaged. Semen was packaged with 35ml per tube creating ~350 individual doses of semen. Every fifth dose was labeled with consecutive numbers (~70/treatment) and overnighted to Purdue University. Concentration of each dose was determined using the NucleoCounter® SP-100TM. The entire study was repeated over 6 weeks. Within treatment, concentration did not differ among doses for the PM and SM treatments. However, NM doses were less concentrated (P< 0.001) in doses >300 compared to all others and doses 251–300 were less concentrated than doses 1 to 50. NM samples 1–250 averaged 36.12 million cells/ml whereas samples 251 to the end of the batch averaged 34.68 million/ml. This results in the last 20 doses having ~50.4 million less cells/dose. Across treatments, NM had lower concentrations (P=0.02) from doses >200 compared to the SM treatment. PM appeared to be an intermediate as it was numerically lower than SM, but not statistically different from either NM or SM. The difference in concentrations between the NM and SM >200 was 1.85 million/ml equaling 64.75 million fewer cells. Overall, allowing semen to settle and not mixing prior to packaging can result in 50–64 million cells fewer per dose in the last 50–100 doses packaged.

中文翻译：

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PSVI-9 自动包装前混合精液对公猪精液的影响

众所周知，精子细胞会随着时间的推移从稀释剂中沉淀出来，这可能会影响批次中每剂量的浓度。本研究的目的是调查包装前混合对包装授精剂量浓度的影响。在商业公猪站，三个精液被收集、扩展、汇集并分配到三个处理组之一：pour-mix (PM)，其中精液在合并三个精液后立即包装，搅拌混合 (SM)，其中三个精液混合并充分搅拌然后包装，或不混合（NM），让汇集的精液静置 10 分钟，然后包装。精液包装为每管 35 毫升，产生约 350 次单独剂量的精液。每第五剂都标有连续数字（~70/治疗），并在普渡大学过夜。使用 NucleoCounter® SP-100TM 确定每个剂量的浓度。整个研究重复了 6 周。在治疗中，PM 和 SM 治疗剂量之间的浓度没有差异。然而，与所有其他剂量相比，NM 剂量在大于 300 的剂量中浓度较低（P<0.001），并且剂量 251-300 的浓度低于剂量 1 至 50。NM 样品 1-250 平均为 3612 万个细胞/ml，而样品 251 至批次末平均3468万/ml。这导致最后 20 个剂量的细胞/剂量减少了约 5040 万个。在处理中，与 SM 处理相比，NM 在大于 200 的剂量下具有较低的浓度 (P=0.02)。PM 似乎是一个中间值，因为它在数值上低于 SM，但与 NM 或 SM 没有统计学差异。NM 和 SM 之间的浓度差异＞200 为 185 万/ml，相当于减少了 6475 万个细胞。总体而言，在包装前允许精液沉淀而不是混合可以导致在最后包装的 50-100 剂中每剂减少 50-6400 万个细胞。