脫脂奶粉和大豆卵磷脂對綿羊精液低溫保存效果的影響
發(fā)布時間：2018-09-05 20:51
【摘要】：本試驗探討了添加脫脂奶粉替代鮮牛奶和大豆卵磷脂替代卵黃在精液低溫保存的可行性,旨在開發(fā)適宜在養(yǎng)羊研究及生產(chǎn)實踐中應(yīng)用的綿羊精液低溫保存稀釋液。假陰道法采集綿羊精液,用不同濃度的脫脂奶粉和大豆卵磷脂分別替代綿羊精液稀釋液中的鮮牛奶和卵黃,對綿羊精液稀釋并進行低溫保存,定時復(fù)溫并對綿羊精子的活力、存活時間、活精子百分率、畸形率和頂體完整率進行檢測,從而評價綿羊精液的低溫保存效果,并將低溫保存稀釋液用于成年綿羊體外受精試驗進行驗證。在綿羊精液稀釋中添加不同濃度的(4%、7%、10%、13%)脫脂奶粉替代鮮牛奶作為試驗組,對照組則添加牛奶,試驗結(jié)果表明:精子活力變化情況,在0~60h各脫脂奶粉組稀釋液與牛奶組無顯著差異(P0.05),但10%脫脂奶粉組高于其他脫脂奶粉組,在72~84h時,10%脫脂奶粉組與牛奶組無顯著性差異(P0.05),而4%、13%脫脂奶粉組顯著低于牛奶組(P0.05),在72~132h,其他脫脂奶粉組(4%、7%、13%)均低于10%脫脂奶粉組和牛奶組(P0.05);活精子百分率變化情況,在0~72h各個脫脂奶粉組均與牛奶組無顯著性差異(P0.05),到96h,10%脫脂奶粉組最高,達到62.62%,與牛奶組差異不顯著(P0.05),120h時,10%脫脂奶粉組與牛奶組無顯著性差異(P0.05),但是顯著高于其他脫脂奶粉組(P0.05);精子畸形率變化情況,在24h、72h和96h三個時間點,7%、10%脫脂奶粉組與牛奶組相互之間差異不顯著(P0.05),10%脫脂奶粉組畸形率最低,在120h時,10%脫脂奶粉組與牛奶組差異不顯著(P0.05),并顯著低于其他脫脂奶粉組(P0.05);精子頂體完整率變化情況,在48h時10%脫脂奶粉組與牛奶組差異不顯著(P0.05),顯著高于4%和13%脫脂奶粉組(P0.05)。綜合以上各指標(biāo)檢測結(jié)果,以10%脫脂奶粉替代鮮牛奶的效果為最佳。在以10%脫脂奶粉替代鮮牛奶進行優(yōu)化綿羊低溫精液稀釋液配方的基礎(chǔ)上,利用不同濃度(0.375%、0.750%、1.500%、3.000%)的大豆卵磷脂取代傳統(tǒng)稀釋液中的卵黃,試驗結(jié)果表明:精子活力變化情況,在0~120h各時間點,0.375%大豆卵磷脂組精子活力與卵黃組差異均不顯著(P0.05),其中在84h時,0.375%大豆卵磷脂組精子活力還維持在0.43,在60~96h,1.500%、3.000%大豆卵磷脂組活力均顯著低于卵黃組和0.375%大豆卵磷脂組(P0.05),在108h時,0.750%、1.500%、3.000%大豆卵磷脂組活力均顯著低于卵黃組(P0.05);精子存活時間情況,0.375%大豆卵磷脂的有效存活時間為88h,總存活時間為120h,與卵黃組組差異不顯著(P0.05),其余各大豆卵磷脂組的有效存活時間和總存活時間均顯著低于0.375%大豆卵磷脂組和卵黃組(P0.05);活精子百分率變化情況,在0、24h時間點,各個組間無顯著性差異(P0.05),從48h~120h,0.375%大豆卵磷脂組與卵黃組差異不顯著(P0.05),尤其在72h、96h、120h,0.375%大豆卵磷脂組與卵黃組差異不顯著(P0.05),但顯著高于0.750%、1.500%、3.000%大豆卵磷脂組(P0.05);精子畸形率變化情況,0~120h,0.375%大豆卵磷脂組與卵黃組差異均不顯著(P0.05),在72h、96h時,0.375%大豆卵磷脂組畸形率最低;精子頂體完整率變化情況,0~120h,各個大豆卵磷脂組均與卵黃組差異不顯著(P0.05)。綜合以上結(jié)果可以看出,0.375%的大豆卵磷脂組替代卵黃效果最好。利用最優(yōu)化的綿羊精液低溫保存稀釋液(10%脫脂奶粉替代新鮮牛奶,0.375%大豆卵磷脂替代卵黃)低溫保存綿羊精精液,進行綿羊的體外受精試驗,以冷凍精液做為對照,結(jié)果表明:低溫組精液的綿羊卵母細胞體外受精卵裂率與冷凍組無顯著性差異(P0.05),但低溫組卵裂率(63.12%)高于冷凍組(55.92%),低溫組的囊胚發(fā)育率(20.61%)顯著高于冷凍組(14.27%)(P0.05);囊胚經(jīng)熒光染色后計數(shù)細胞數(shù),低溫組(111.64)和冷凍組(109.60)無顯著差異(P0.05)。
[Abstract]:The feasibility of using skimmed milk powder instead of fresh milk and soybean lecithin instead of yolk in cryopreservation of semen was studied in this experiment. The purpose of this experiment was to develop Sheep Semen Cryopreservation dilution suitable for sheep breeding research and production practice. The fresh milk and egg yolk in sheep semen dilution solution were diluted and cryopreserved, and the sperm viability, survival time, percentage of live sperm, deformity rate and acrosome integrity of sheep were detected. The cryopreserved semen dilution was used in adult sheep. External fertilization test was carried out to verify the results. Different concentrations of skimmed milk powder (4%, 7%, 10%, 13%) were added in the dilution of Sheep Semen to replace fresh milk as the experimental group, while milk was added in the control group. The results showed that there was no significant difference in sperm motility between the skimmed milk powder group and the milk group at 0-60 h (P 0.05), but the 10% skimmed milk powder group was higher than the milk group. Other skimmed milk powder groups, at 72-84 hours, 10% skimmed milk powder group and milk group had no significant difference (P 0.05), but 4%, 13% skimmed milk powder group was significantly lower than milk group (P 0.05), at 72-132 hours, other skimmed milk powder group (4%, 7%, 13%) were lower than 10% skimmed milk powder group and milk group (P 0.05); the percentage of live sperm was changed in each skimmed milk powder group at 0-72 hours. There was no significant difference between the milk group and the 10% skimmed milk powder group (P 0.05). At 96 h, the 10% skimmed milk powder group was the highest, reaching 62.62%, which was not significantly different from the milk group (P 0.05). At 120 h, the 10% skimmed milk powder group and the milk group had no significant difference (P 0.05), but significantly higher than the other skimmed milk powder group (P 0.05); the change of sperm deformity rate was 7%, 10% skimmed at 24 h, 72 h and 96 h. There was no significant difference between the fat milk powder group and the milk group (P 0.05). The deformity rate of the 10% skimmed milk powder group was the lowest. At 120 hours, the deformity rate of the 10% skimmed milk powder group and the milk group was not significantly different (P 0.05), and was significantly lower than that of the other skimmed milk powder groups (P 0.05). It was significantly higher than that of 4% and 13% skimmed milk powder group (P 0.05). According to the above results, 10% skimmed milk powder was the best substitute for fresh milk. The results showed that the sperm motility of 0.375% soybean lecithin group was not significantly different from that of egg yolk group (P 0.05) at 0-120 H. The sperm motility of 0.375% soybean lecithin group remained at 0.43, 60-96 h, 1.500%, 3.000% soybean lecithin group was significantly lower than that of egg at 84 H. The activity of 0.750%, 1.500% and 3.000% soybean lecithin group was significantly lower than that of lecithin group (P 0.05) at 108 hours in yellow group and 0.375% soybean lecithin group (P 0.05); the effective survival time of sperm in 0.375% soybean lecithin group was 88 hours and the total survival time was 120 hours, which was not significantly different from that in yolk group (P 0.05). Survival time and total survival time were significantly lower than 0.375% soybean lecithin group and yolk group (P 0.05); the percentage of live sperm had no significant difference (P 0.05) between each group at 0 and 24 hours, and there was no significant difference (P 0.05) between soybean lecithin group and yolk group from 48 to 120 hours, 0.375% soybean lecithin group and yolk group (P 0.05), especially at 72 hours, 96 hours, 120 hours, 0.375% soybean lecithin group and egg yolk group. There was no significant difference between yellow group and yolk group (P 0.05), but it was significantly higher than 0.750%, 1.500%, 3.000% soybean lecithin group (P 0.05); sperm deformity rate was not significantly different between 0-120 h, 0.375% soybean lecithin group and yolk group (P 0.05); at 72 h, 96 h, 0.375% soybean lecithin group had the lowest deformity rate; acrosome integrity rate of sperm, 0-120 h, each soybean egg. The results showed that 0.375% soybean lecithin group had the best effect on replacing yolk. Sheep semen was cryopreserved in vitro with the optimum dilution of Sheep Semen (10% skim milk instead of fresh milk, 0.375% soybean lecithin instead of yolk). The results showed that there was no significant difference in cleavage rate between cryogenic group and cryogenic group (P 0.05), but the cleavage rate of cryogenic group (63.12%) was higher than that of cryogenic group (55.92%). The blastocyst development rate of cryogenic group (20.61%) was significantly higher than that of cryogenic group (14.27%) (P 0.05). The number of counted cells in cryogenic group (111.64) and cryopreservation group (109.60) had no significant difference (P0.05).
【學(xué)位授予單位】：河北農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S826
本文編號：2225429
[Abstract]:The feasibility of using skimmed milk powder instead of fresh milk and soybean lecithin instead of yolk in cryopreservation of semen was studied in this experiment. The purpose of this experiment was to develop Sheep Semen Cryopreservation dilution suitable for sheep breeding research and production practice. The fresh milk and egg yolk in sheep semen dilution solution were diluted and cryopreserved, and the sperm viability, survival time, percentage of live sperm, deformity rate and acrosome integrity of sheep were detected. The cryopreserved semen dilution was used in adult sheep. External fertilization test was carried out to verify the results. Different concentrations of skimmed milk powder (4%, 7%, 10%, 13%) were added in the dilution of Sheep Semen to replace fresh milk as the experimental group, while milk was added in the control group. The results showed that there was no significant difference in sperm motility between the skimmed milk powder group and the milk group at 0-60 h (P 0.05), but the 10% skimmed milk powder group was higher than the milk group. Other skimmed milk powder groups, at 72-84 hours, 10% skimmed milk powder group and milk group had no significant difference (P 0.05), but 4%, 13% skimmed milk powder group was significantly lower than milk group (P 0.05), at 72-132 hours, other skimmed milk powder group (4%, 7%, 13%) were lower than 10% skimmed milk powder group and milk group (P 0.05); the percentage of live sperm was changed in each skimmed milk powder group at 0-72 hours. There was no significant difference between the milk group and the 10% skimmed milk powder group (P 0.05). At 96 h, the 10% skimmed milk powder group was the highest, reaching 62.62%, which was not significantly different from the milk group (P 0.05). At 120 h, the 10% skimmed milk powder group and the milk group had no significant difference (P 0.05), but significantly higher than the other skimmed milk powder group (P 0.05); the change of sperm deformity rate was 7%, 10% skimmed at 24 h, 72 h and 96 h. There was no significant difference between the fat milk powder group and the milk group (P 0.05). The deformity rate of the 10% skimmed milk powder group was the lowest. At 120 hours, the deformity rate of the 10% skimmed milk powder group and the milk group was not significantly different (P 0.05), and was significantly lower than that of the other skimmed milk powder groups (P 0.05). It was significantly higher than that of 4% and 13% skimmed milk powder group (P 0.05). According to the above results, 10% skimmed milk powder was the best substitute for fresh milk. The results showed that the sperm motility of 0.375% soybean lecithin group was not significantly different from that of egg yolk group (P 0.05) at 0-120 H. The sperm motility of 0.375% soybean lecithin group remained at 0.43, 60-96 h, 1.500%, 3.000% soybean lecithin group was significantly lower than that of egg at 84 H. The activity of 0.750%, 1.500% and 3.000% soybean lecithin group was significantly lower than that of lecithin group (P 0.05) at 108 hours in yellow group and 0.375% soybean lecithin group (P 0.05); the effective survival time of sperm in 0.375% soybean lecithin group was 88 hours and the total survival time was 120 hours, which was not significantly different from that in yolk group (P 0.05). Survival time and total survival time were significantly lower than 0.375% soybean lecithin group and yolk group (P 0.05); the percentage of live sperm had no significant difference (P 0.05) between each group at 0 and 24 hours, and there was no significant difference (P 0.05) between soybean lecithin group and yolk group from 48 to 120 hours, 0.375% soybean lecithin group and yolk group (P 0.05), especially at 72 hours, 96 hours, 120 hours, 0.375% soybean lecithin group and egg yolk group. There was no significant difference between yellow group and yolk group (P 0.05), but it was significantly higher than 0.750%, 1.500%, 3.000% soybean lecithin group (P 0.05); sperm deformity rate was not significantly different between 0-120 h, 0.375% soybean lecithin group and yolk group (P 0.05); at 72 h, 96 h, 0.375% soybean lecithin group had the lowest deformity rate; acrosome integrity rate of sperm, 0-120 h, each soybean egg. The results showed that 0.375% soybean lecithin group had the best effect on replacing yolk. Sheep semen was cryopreserved in vitro with the optimum dilution of Sheep Semen (10% skim milk instead of fresh milk, 0.375% soybean lecithin instead of yolk). The results showed that there was no significant difference in cleavage rate between cryogenic group and cryogenic group (P 0.05), but the cleavage rate of cryogenic group (63.12%) was higher than that of cryogenic group (55.92%). The blastocyst development rate of cryogenic group (20.61%) was significantly higher than that of cryogenic group (14.27%) (P 0.05). The number of counted cells in cryogenic group (111.64) and cryopreservation group (109.60) had no significant difference (P0.05).
【學(xué)位授予單位】：河北農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S826
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