日粮蛋白质和蛋氨酸水平对水貂生产性能及毛皮发育的影响
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摘要
本论文通过研究水貂日粮适宜可消化蛋白质和蛋氨酸水平,结合日粮蛋白质和蛋氨酸水平对水貂生产性能、毛囊发育和水貂皮肤内参与水貂皮肤和毛囊生长相关基因的定量定位表达研究,分别从营养水平、组织形态学水平和分子水平对日粮蛋白质和蛋氨酸对水貂毛皮性能影响机理进行了探索性研究,主要内容如下:
1.选取健康断奶公水貂128只,随机分成8组,每组16只幼貂,各组水貂初始体重差异不显著。前5组分别饲喂日粮粗蛋白质水平为36%、32%、28%、24%和20%的日粮,后三组在日粮蛋白质水平为28%的日粮中分别添加0.3%、0.6%和0.9%的DL-蛋氨酸,试验期60天。结果显示:当日粮蛋白质水平降低到28%时添加0.6%蛋氨酸日粮组水貂的试验末体重、日增重、饲料转化率及氮沉积均最佳,且显著的高于36%的高蛋白质日粮组。
结论:育成期水貂可消化蛋白质水平为24.0%,日粮代谢能为20.73 MJ/kg,日粮蛋氨酸水平为1.39%,总含硫氨基酸水平为2.70%,水貂可获得最佳生产性能。
2.在水貂育成期试验基础上,每组留取14只健康公水貂,总计112只,各组水貂初始体重差异不显著。前五组分别饲喂日粮粗蛋白质水平为32%、28%、24%、20%和16%的日粮,后三组在日粮蛋白质水平为24%的日粮中分别添加0.4%、0.8%和1.2%的DL-蛋氨酸,试验期85天。结果显示:当日粮蛋白质水平降至24%补充0.8%的蛋氨酸时,水貂生产性能有提高的趋势,且能够使水貂试验末体重、氮沉积和干皮长均达到最大。
结论:冬毛期水貂可消化蛋白质水平为19.7%,日粮代谢能为21.66MJ /kg,日粮中蛋氨酸水平为1.54%,总含硫氨基酸水平为2.82%,水貂可获得最佳生产性能。
3.分别从饲喂日粮蛋白质水平为32%、24%、16%和24%+0.8%蛋氨酸的处理组,每组采集
4只水貂皮肤样品进行石蜡切片。结果显示:水貂毛囊多为三毛型,初级毛囊有规律的排列在毛囊群的一侧,冬毛期水貂次级毛囊与初级毛囊的平均比例(S/P)为20~22:1;水貂皮肤经Sacpic法染色后,将有活性的毛囊内根鞘染成红色;高蛋白质水平和适宜的蛋氨酸水平日粮组水貂针绒毛活性低。
结论:Sacpic染色法适宜水貂皮肤的染色;针毛和绒毛毛囊的活性周期不同步;高蛋白质水平和适宜蛋氨酸水平日粮对水貂毛囊发育有正向调节作用;水貂打皮季节其毛囊正处于退行期。
4.分别从饲喂蛋白质水平为32%、24%、16%和24%+0.8%蛋氨酸的处理组,每组采集4只水貂皮肤样品,对水貂皮肤中IGF-Ⅰ、IGF-ⅠR和EGF mRNA进行相对定量研究。结果显示:随着日粮蛋白质水平的降低,各基因mRNA的表达量有显著的下降趋势,当低蛋白质日粮中添加蛋氨酸后,可以提高各基因mRNA的表达量;IGF-Ⅰ、IGF-ⅠR基因的表达量与水貂的针毛长度、绒毛长度、S/P等毛皮性能指标有显著的相关性,EGF和IGF-ⅠR的表达量有呈显著的正关性。
结论:高蛋白质水平和适宜蛋氨酸水平日粮对水貂皮肤中IGF-Ⅰ、IGF-ⅠR和EGF基因的表达量有显著的提高作用;IGF-Ⅰ与IGF-ⅠR基因表达量与水貂毛皮性能指标存在显著的相关关系。
5.通过冰冻切片和免疫组织化学方法对水貂皮肤中的EGF、IGF-Ⅰ及IGF-ⅠR基因的分布规律进行了研究。结果显示:处于打皮季节的水貂,皮肤中IGF-Ⅰ、IGF-ⅠR和EGF基因广泛分布在毛囊外根鞘细胞和皮肤表皮细胞的胞质中。
结论:IGF-Ⅰ、IGF-ⅠR和EGF基因在水貂皮肤表皮和毛囊外根鞘细胞中广泛表达,直接参与调控皮肤和毛囊的发育,日粮蛋白质和蛋氨酸通过调控水貂皮肤内IGF-Ⅰ、IGF-ⅠR和EGF基因的表达水平,从而作为影响水貂毛皮品质的一个重要途径。
The paper studied on the protein and methionine (Met) level of minks’diet, and worked on the performance, hair follicle development of the mink and some important growth factors which had special functions in skin and hair follicle growth. From nutrition, microstructure morphological and molecular aspects, respectively, to study how the protein and Met affected the fur characters of mink.
Main contents were as follows:
1. 128 healthy male minks were selected and randomly divided into eight groups with different types of diet, average body weight was similar in all groups. The first five groups dietary protein level were 36%, 32%, 28%, 24% and 20%, the last three groups were supplemented with Met (0.3%, 0.6% and 0.9% DM) in 28% diet, the experiment lasted for 60 days. The results showed that: minks in 28% protein level diet with 0.6% Met group obtained the best body weight, average daily gain, F/G and the daily retention of nitrogen.
Conclusion: the digestible protein level of growing minks’diet was 24.0%, the ME of diet was 20.73MJ/kg and the level of Met in diet was 1.39%, sulphur containing amino acids in diet was 2.70%, minks obtained the best performance.
2. Based on the growing period experiment, each group left 14 healthy minks, total was 112, and average body weight was similar in all groups. The first five groups dietary protein level were 32%, 28%, 24%, 20% and 16%, the last three groups were supplemented with Met (0.4%, 0.8% and 1.2% DM) in 24% diet, the experiment lasted for 85 days. The results showed that: minks in 24%+0.8% Met group obtained the maximized body weight, daily retention of nitrogen and dry fur length.
Conclusion: the digestible protein level of diet was 19.7%, the ME of diet was 21.66MJ/kg, the level of Met in diet was 1.54%, and sulphur containing amino acids in diet was 2.82%, minks obtained the best performance.
3. We studied on the mink’s skin from groups 32%, 24%, 16% and 24%+0.8 Met by paraffin slice. The result showed that: mink’s hair follicles mostly have three primary hair follicles, and they arranged regularly in the same side of the secondary hair follicles. Secondary hair follicles and primary hair follicles average ratio (S/P) was 20~22:1 in the pelting period of minks. Being dyed with Sacpic method, the inner root sheath was dyed red if it was an activated hair follicle. High protein level and optimal Met level diets promoted the hair follicles to the downward period of activity cycle simultaneously.
Conclusion: Sacpic dyeing method was suitable for mink’s skin dyeing; the activity of primary hair follicles and secondary hair follicles were not synchronizated in hair follicles activity cycle; dietary protein and Met levels affected the hair follicles development.
4. We studied on the growth factor gene expressions, which were relevant mink’s skin and fur growth from groups 32%, 24%, 16% and 24%+0.8 Met. We used real-time PCR technology to study the gene expression quantity of IGF-Ⅰ, IGF-ⅠR and EGF mRNA in mink’s skin. The result showed that each gene mRNA expression quantity decreased with the declining dietary protein levels, the expressions of each gene mRNA improved when the low-protein diet was supplemented with Met. The quantity of IGF-Ⅰand IGF-ⅠR mRNA expressions significantly correlative with the length of the guard hair, under hair and S/P of the minks. EGF gene expression was correlative with the IGF-ⅠR mRNA expressions.
Conclusion: High dietary protein level and optimal Met level diets improved the quantity of IGF-Ⅰand IGF-ⅠR mRNA expressions. The quantity of IGF-Ⅰand IGF-ⅠR mRNA expressions were significantly correlative with some skin and fur performance characters of the minks. 5. We studied on the position of IGF-Ⅰ, IGF-ⅠR and EGF in mink’s skin by frozen section and immunohistochemical method. The result showed that: IGF-Ⅰ, IGF-ⅠR and EGF mainly distributed in the cell cytoplasm of skin epidermis cells and out root sheath cells.
Conclusion: dietary protein and Met influenced the IGF-Ⅰ, IGF-ⅠR and EGF mRNA expressions, these genes played a role in skin epidermis cells and out root sheath cells in mink’s skin and then affected the skin growth, hair follicles and fur characteristics of minks.
1.选取健康断奶公水貂128只,随机分成8组,每组16只幼貂,各组水貂初始体重差异不显著。前5组分别饲喂日粮粗蛋白质水平为36%、32%、28%、24%和20%的日粮,后三组在日粮蛋白质水平为28%的日粮中分别添加0.3%、0.6%和0.9%的DL-蛋氨酸,试验期60天。结果显示:当日粮蛋白质水平降低到28%时添加0.6%蛋氨酸日粮组水貂的试验末体重、日增重、饲料转化率及氮沉积均最佳,且显著的高于36%的高蛋白质日粮组。
结论:育成期水貂可消化蛋白质水平为24.0%,日粮代谢能为20.73 MJ/kg,日粮蛋氨酸水平为1.39%,总含硫氨基酸水平为2.70%,水貂可获得最佳生产性能。
2.在水貂育成期试验基础上,每组留取14只健康公水貂,总计112只,各组水貂初始体重差异不显著。前五组分别饲喂日粮粗蛋白质水平为32%、28%、24%、20%和16%的日粮,后三组在日粮蛋白质水平为24%的日粮中分别添加0.4%、0.8%和1.2%的DL-蛋氨酸,试验期85天。结果显示:当日粮蛋白质水平降至24%补充0.8%的蛋氨酸时,水貂生产性能有提高的趋势,且能够使水貂试验末体重、氮沉积和干皮长均达到最大。
结论:冬毛期水貂可消化蛋白质水平为19.7%,日粮代谢能为21.66MJ /kg,日粮中蛋氨酸水平为1.54%,总含硫氨基酸水平为2.82%,水貂可获得最佳生产性能。
3.分别从饲喂日粮蛋白质水平为32%、24%、16%和24%+0.8%蛋氨酸的处理组,每组采集
4只水貂皮肤样品进行石蜡切片。结果显示:水貂毛囊多为三毛型,初级毛囊有规律的排列在毛囊群的一侧,冬毛期水貂次级毛囊与初级毛囊的平均比例(S/P)为20~22:1;水貂皮肤经Sacpic法染色后,将有活性的毛囊内根鞘染成红色;高蛋白质水平和适宜的蛋氨酸水平日粮组水貂针绒毛活性低。
结论:Sacpic染色法适宜水貂皮肤的染色;针毛和绒毛毛囊的活性周期不同步;高蛋白质水平和适宜蛋氨酸水平日粮对水貂毛囊发育有正向调节作用;水貂打皮季节其毛囊正处于退行期。
4.分别从饲喂蛋白质水平为32%、24%、16%和24%+0.8%蛋氨酸的处理组,每组采集4只水貂皮肤样品,对水貂皮肤中IGF-Ⅰ、IGF-ⅠR和EGF mRNA进行相对定量研究。结果显示:随着日粮蛋白质水平的降低,各基因mRNA的表达量有显著的下降趋势,当低蛋白质日粮中添加蛋氨酸后,可以提高各基因mRNA的表达量;IGF-Ⅰ、IGF-ⅠR基因的表达量与水貂的针毛长度、绒毛长度、S/P等毛皮性能指标有显著的相关性,EGF和IGF-ⅠR的表达量有呈显著的正关性。
结论:高蛋白质水平和适宜蛋氨酸水平日粮对水貂皮肤中IGF-Ⅰ、IGF-ⅠR和EGF基因的表达量有显著的提高作用;IGF-Ⅰ与IGF-ⅠR基因表达量与水貂毛皮性能指标存在显著的相关关系。
5.通过冰冻切片和免疫组织化学方法对水貂皮肤中的EGF、IGF-Ⅰ及IGF-ⅠR基因的分布规律进行了研究。结果显示:处于打皮季节的水貂,皮肤中IGF-Ⅰ、IGF-ⅠR和EGF基因广泛分布在毛囊外根鞘细胞和皮肤表皮细胞的胞质中。
结论:IGF-Ⅰ、IGF-ⅠR和EGF基因在水貂皮肤表皮和毛囊外根鞘细胞中广泛表达,直接参与调控皮肤和毛囊的发育,日粮蛋白质和蛋氨酸通过调控水貂皮肤内IGF-Ⅰ、IGF-ⅠR和EGF基因的表达水平,从而作为影响水貂毛皮品质的一个重要途径。
The paper studied on the protein and methionine (Met) level of minks’diet, and worked on the performance, hair follicle development of the mink and some important growth factors which had special functions in skin and hair follicle growth. From nutrition, microstructure morphological and molecular aspects, respectively, to study how the protein and Met affected the fur characters of mink.
Main contents were as follows:
1. 128 healthy male minks were selected and randomly divided into eight groups with different types of diet, average body weight was similar in all groups. The first five groups dietary protein level were 36%, 32%, 28%, 24% and 20%, the last three groups were supplemented with Met (0.3%, 0.6% and 0.9% DM) in 28% diet, the experiment lasted for 60 days. The results showed that: minks in 28% protein level diet with 0.6% Met group obtained the best body weight, average daily gain, F/G and the daily retention of nitrogen.
Conclusion: the digestible protein level of growing minks’diet was 24.0%, the ME of diet was 20.73MJ/kg and the level of Met in diet was 1.39%, sulphur containing amino acids in diet was 2.70%, minks obtained the best performance.
2. Based on the growing period experiment, each group left 14 healthy minks, total was 112, and average body weight was similar in all groups. The first five groups dietary protein level were 32%, 28%, 24%, 20% and 16%, the last three groups were supplemented with Met (0.4%, 0.8% and 1.2% DM) in 24% diet, the experiment lasted for 85 days. The results showed that: minks in 24%+0.8% Met group obtained the maximized body weight, daily retention of nitrogen and dry fur length.
Conclusion: the digestible protein level of diet was 19.7%, the ME of diet was 21.66MJ/kg, the level of Met in diet was 1.54%, and sulphur containing amino acids in diet was 2.82%, minks obtained the best performance.
3. We studied on the mink’s skin from groups 32%, 24%, 16% and 24%+0.8 Met by paraffin slice. The result showed that: mink’s hair follicles mostly have three primary hair follicles, and they arranged regularly in the same side of the secondary hair follicles. Secondary hair follicles and primary hair follicles average ratio (S/P) was 20~22:1 in the pelting period of minks. Being dyed with Sacpic method, the inner root sheath was dyed red if it was an activated hair follicle. High protein level and optimal Met level diets promoted the hair follicles to the downward period of activity cycle simultaneously.
Conclusion: Sacpic dyeing method was suitable for mink’s skin dyeing; the activity of primary hair follicles and secondary hair follicles were not synchronizated in hair follicles activity cycle; dietary protein and Met levels affected the hair follicles development.
4. We studied on the growth factor gene expressions, which were relevant mink’s skin and fur growth from groups 32%, 24%, 16% and 24%+0.8 Met. We used real-time PCR technology to study the gene expression quantity of IGF-Ⅰ, IGF-ⅠR and EGF mRNA in mink’s skin. The result showed that each gene mRNA expression quantity decreased with the declining dietary protein levels, the expressions of each gene mRNA improved when the low-protein diet was supplemented with Met. The quantity of IGF-Ⅰand IGF-ⅠR mRNA expressions significantly correlative with the length of the guard hair, under hair and S/P of the minks. EGF gene expression was correlative with the IGF-ⅠR mRNA expressions.
Conclusion: High dietary protein level and optimal Met level diets improved the quantity of IGF-Ⅰand IGF-ⅠR mRNA expressions. The quantity of IGF-Ⅰand IGF-ⅠR mRNA expressions were significantly correlative with some skin and fur performance characters of the minks. 5. We studied on the position of IGF-Ⅰ, IGF-ⅠR and EGF in mink’s skin by frozen section and immunohistochemical method. The result showed that: IGF-Ⅰ, IGF-ⅠR and EGF mainly distributed in the cell cytoplasm of skin epidermis cells and out root sheath cells.
Conclusion: dietary protein and Met influenced the IGF-Ⅰ, IGF-ⅠR and EGF mRNA expressions, these genes played a role in skin epidermis cells and out root sheath cells in mink’s skin and then affected the skin growth, hair follicles and fur characteristics of minks.
引文
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