Department of Veterinary Medicine and Animal Science, University of Milan, 26900, Lodi, Italy.
Department of Agricultural, Environmental and Food Science, University of Molise, 86100, Campobasso, Italy.
Cryobiology. 2022 Jun;106:66-72. doi: 10.1016/j.cryobiol.2022.04.001. Epub 2022 Apr 13.
The aim of this study was to compare the effect of two permeant-cryoprotectants, dimethylacetamide (DMA) and N-methylacetamide (NMA) used at different concentrations (0%, 2%, 4%, 6%) on the quality and fertility of post-thaw rooster semen. Ejaculates were processed in 7 treatments: Lake pre-freezing+0.1 M trehalose (LPF-T) (control treatment), LPF-T+2% DMA, LPF-T+4% DMA, LPF-T+6% DMA, LPF-T+2% NMA, LPF-T+4% NMA, LPF-T+6% NMA. Sperm quality [sperm membrane integrity (SMI), motility and kinetic parameters] was assessed before and after cryopreservation. Fertility and embryo viability were recorded. Increasing both DMA and NMA concentration from 2 to 6% improved SMI, total motile sperm, progressive motile sperm (PMS), VCL, VSL and VAP values. PMS recovery rates were significantly the highest in 6% DMA, 4% NMA and 6% NMA treatments. Semen cryopreserved with DMA produced the best fertility and embryo viability at 6%; progressive lower values were recorded at lower concentrations, with no viable embryos at 2%. Semen cryopreserved with NMA showed the best fertility values at 2% and lower values were recorded at higher concentrations; live embryos were found in all NMA treatments. Finally, NMA and DMA showed a similar positive concentration dependent effect of the quality of cryopreserved semen. NMA, not DMA, provided the highest fertility and embryo viability values at the lowest 2%. Therefore, the use of NMA is recommended in order to reduce the cryoprotectant concentration, with a concomitant reduction in the risk of toxicity, providing at the same time the adequate cryoprotective action to obtain viable embryos after artificial insemination of cryopreserved chicken semen.
本研究旨在比较两种透性冷冻保护剂二甲基乙酰胺(DMA)和 N-甲基乙酰胺(NMA)在不同浓度(0%、2%、4%、6%)下对解冻后公鸡精液质量和生育力的影响。精液分别在 7 种处理中进行处理:湖预冻+0.1 M海藻糖(LPF-T)(对照处理)、LPF-T+2% DMA、LPF-T+4% DMA、LPF-T+6% DMA、LPF-T+2% NMA、LPF-T+4% NMA、LPF-T+6% NMA。在冷冻保存前后评估精子质量[精子膜完整性(SMI)、活力和运动参数]。记录受精率和胚胎存活率。DMA 和 NMA 浓度从 2%增加到 6%均提高了 SMI、总运动精子数、前向运动精子数(PMS)、VCL、VSL 和 VAP 值。6% DMA、4% NMA 和 6% NMA 处理中的 PMS 回收率显著最高。用 DMA 冷冻保存的精液在 6%时产生最佳的受精率和胚胎存活率;浓度越低,值越低,在 2%时没有存活的胚胎。用 NMA 冷冻保存的精液显示出最佳的受精率,在 2%时,浓度越高,值越低;在所有 NMA 处理中都发现了活胚胎。最后,NMA 和 DMA 对冷冻精液质量均表现出相似的正浓度依赖性影响。NMA,而不是 DMA,在最低 2%时提供了最高的受精率和胚胎存活率。因此,为了降低冷冻保护剂的浓度,同时降低毒性风险,建议使用 NMA,以获得在冷冻鸡精液人工授精后可存活的胚胎。