【作者】 胡骏鹏；

【导师】 何瑞国； 熊远著；

【作者基本信息】 华中农业大学 ， 动物遗传育种与繁殖， 2008， 博士

【摘要】 朗德鹅和溆浦鹅是国内主要的鹅肥肝品种。在肥肝鹅产业化开发过程中,品系培育是其中的一个重要方面。大多数研究表明,日粮ME水平对早期鹅增重的影响比蛋白质水平影响大,但少有研究者就能量一个因素对肥肝鹅品系的早期生长发育进行特定研究。本课题通过改变日粮代谢能,结合朗德鹅和溆浦鹅早期生长情况、关键日龄屠体指标、相关血液生化指标和血清激素水平和肌肉生成抑制因子(MSTN)基因表达量等因素,探索朗德鹅和溆浦鹅早期生长发育规律、超饲养前期所需要的适宜ME营养需要和涉及早期生长发育的相关规律。试验结果分别摘要如下:试验1 0～70d阶段能量对朗德鹅公鹅生长发育影响的研究目的:研究不同日粮能量水平对朗德鹅公鹅超饲养前期(0～70d)生长性能指标、屠体性能、体尺指标、血液生化指标、激素水平和肌肉成分的影响研究。方法:本试验用90只0日龄初生重大致相同的朗德鹅公鹅为试验对象,通过添加豆油来设计不高中低三种不同代谢能水平(0～3w:13.38MJ/kg、12.13MJ/kg和10.87MJ/kg;4～10w:13.81MJ/kg、12.55MJ/kg和11.30MJ/kg)的日粮,试验鹅随机分为试验Ⅰ组,试验Ⅱ组,试验Ⅲ组,每组30只,分栏饲养70天。每周进行称重和体尺测定;于21d、70d进行屠宰试验,屠宰前采集血液进行血清参数和血清激素水平检测;屠宰后测定肌肉成分。结果:(1)适宜朗德鹅公鹅能量水平:0～21d为12.13MJ/kg、22～70d为12.55MJ/Kg;(2)日粮能量对于朗德鹅公鹅屠宰率、全净膛率、半净膛率、腿肌率的影响差异均不显著(p＞0.05);(3)朗德鹅公鹅肌肉中的蛋白质含量超过21%;胸肌和腿肌总脂含量都是随着日粮能量浓度提高而提高,Ⅰ组胸肌和腿肌总脂含量比Ⅲ组分别高24.22%和34.73%;胸肌中各项化学成分含量高于腿肌;(4)21d时,朗德鹅公鹅血清中TG、LDL、HLDL差异不显著(p＞0.05);70d时,朗德鹅公鹅血清中AST、LDL和HLDL的含量差异不显著(p＞0.05);(5)21d血清IGF—Ⅰ测定值(♂Ⅲ除外)均高于70d时各组测定值;且ME水平对21d时血清IGF—Ⅰ水平影响差异显著(p＜0.05);21d血清GH测定值随日粮能量的升高而升高,70d时,公鹅各组GH差异不显著(p＞0.05)。试验2 0～70d阶段能量对朗德鹅母鹅生长发育影响的研究目的:研究不同日粮能量水平对朗德鹅母鹅超饲养前期(0～70d)生长性能指标、屠体性能、体尺指标、血液生化指标、激素水平和肌肉成分的影响研究。方法:同朗德鹅公鹅。结果:(1)适宜朗德鹅母鹅能量水平:0～21d为13.38MJ/kg、22～70d为13.81MJ/Kg;(2)各组母鹅腿肌中干物质含量分别要高于同组的胸肌干物质含量约4.58%、0.21%和10.29%;胸肌中总脂含量分别要高于同组的腿肌总脂含量约1.53%、6.91%和1.88%;(3)21d时,朗德鹅母鹅TG、LDL、HLDL差异不显著(p＞0.05);70d时,朗德鹅母鹅血清中AST、LDL、TC、AKP和HLDL的含量差异不显著(p＞0.05)。(4)21d血清IGF—Ⅰ测定值均高于70d测定值;21d时,母鹅IGF—Ⅰ各组差异不显著(p＞0.05),70d差异显著(p＜0.05)。21d时,母鹅GH随日粮能量的提高而提高。试验3 0～70d阶段能量对溆浦鹅生长发育影响的研究目的:研究不同日粮能量水平对溆浦鹅超饲养前期(0～70d)生长性能指标、屠体性能、体尺指标、血液生化指标、激素水平和肌肉成分的影响研究。方法:同朗德鹅公鹅。结果:(1)适宜溆浦鹅能量水平:0～21d能量水平为13.38MJ/kg、22～70d能量水平为13.81MJ/Kg。(2)采食中等能量的溆浦鹅Ⅱ组在屠宰率、全净膛率、半净膛率、腹脂率和胸肌率五个指标上和其余两组差异显著(p＜0.05);对于皮下脂肪厚度和肌间脂肪厚度两个指标上溆浦鹅Ⅰ、Ⅱ组和Ⅲ组之间差异显著(p＜0.05);Ⅱ组的腹脂重和腹脂率高于其余两组,差异显著(p＜0.05)。(3)采食不同能量日粮溆浦鹅胸肌和腿肌蛋白质含量差异不显著(p＞0.05);Ⅰ组胸肌中总脂含量分别比Ⅱ、Ⅲ组胸肌总脂含量高4.46%和8.66%;Ⅰ组腿肌中总脂含量分别比Ⅱ、Ⅲ组腿肌总脂含量高4.99%和11.63%。(4)21d和70d时,溆溆浦鹅血清IGF—Ⅰ测定值差异不显著(p＞0.05);21d时血清GH测定值低于70d时测定值。试验1—3结论:(1)朗德鹅公鹅、朗德鹅母鹅和溆浦鹅早期适宜ME水平:0～21d分别为12.13MJ/kg、13.38MJ/kg和13.38MJ/kg;22～70d分别为12.55MJ/kg、13.81MJ/kg和13.81MJ/kg;(2)本试验中,日粮能量水平可以影响鹅的体尺发育;(3)ME水平显著影响朗德鹅部分屠宰指标(公鹅屠体重、全净膛重、半净膛重、胸肌重、腿肌重和腹脂重);日粮能量水平对朗德鹅屠宰率、全净膛率、半净膛率、腿肌率的影响差异均不显著;(4)ME水平极显著影响溆浦鹅的屠宰率、全净膛率、半净膛率、腹脂率和胸肌率等五个指标;(5)朗德鹅肌肉粗脂肪含量高于溆浦鹅肌肉粗脂肪含量;同等能量水平下朗德鹅公鹅粗蛋白质、粗脂肪和粗灰分含量高于朗德鹅母鹅;ME水平会影响肌肉化学组成;(6)GH水平随ME提高而提高;血清IGF—Ⅰ水平和日粮能量水平无相关性。试验4朗德鹅、溆浦鹅肌肉生成抑制因子(MSTN)基因的克隆和能量对其表达量影响的研究目的:采用反转录—聚合酶链反应(RT—PCR)技术,克隆朗德鹅、溆浦鹅肌肉生成抑制因子(MSTN)基因,研究MSTN基因表达和日粮能量及其部分血清激素含量的相互关系,了解MSTN的功能,为水禽分子营养和分子育种提供基础资料。方法:从朗德鹅、溆浦鹅的腿肌中抽提总RNA,用两步法RT—PCR扩增出MSTN基因的cDNA编码序列,以pGEM—T Vector为载体,将该片段克隆到大肠杆菌(Escherichia coli)中。通过筛选阳性克隆,双酶切鉴定后测序;以MSTN基因的克隆为基础,以β—actin为内标,构建优化的半定量RT—PCR方法,研究三种不同能量对朗德鹅公鹅、朗德鹅母鹅和溆浦鹅在21d和70d两个时期肌肉组织MSTN基因表达的差异;同时用放免法测定21、70d的血清GH、IGF—Ⅰ浓度。结果:(1)克隆到朗德鹅、溆浦鹅MSTN基因cDNA的部分序列,其片段大小为1128bp,编码375个氨基酸组成的多肽,与已发表的鸡、鸭、鹅的MSTN核苷酸相似性分别为94%、94%、99%;氨基酸的相似性分别为98%、97%、98%;(2)21d时,朗德鹅公鹅、母鹅和溆浦鹅MSTN表达量差异不显著;(3)70d时朗德鹅公鹅MSTN的表达量情况为Ⅰ＞Ⅲ＞Ⅱ,朗德鹅母鹅MSTN的表达量情况为Ⅲ＞Ⅱ＞Ⅰ,溆浦鹅MSTN的表达量情况为Ⅲ＞Ⅱ＞Ⅰ;(4)日粮能量对21d后朗德鹅、溆浦鹅MSTN基因的表达有影响,MSTN基因的表达量和血清IGF—Ⅰ的变化趋势基本一致;与血清GH含量无相关性,GH和能量的高低呈正相关。结论:日粮能量对21d后朗德鹅、溆浦鹅MSTN基因的表达有影响,且MSTN基因表达量和血清IGF—Ⅰ具有相关性,和血清GH无相关性。更多还原

【Abstract】 Landes geese and Xupu geese are the main kinds domestic fatty liver geese in China currently.Breeding fatty liver geese is one of the most important aspects in the development process of geese industrialization.Most researchers consider ME levels exert more greater influence on earlier period geese gain than protein level,but up to the present,there are no researchers who research a energy factor to the geese kind specifically.The earlier growth performance rule of landes geese and Xupu geese,the standard of feasible ME nutrition needing beyond earlier period and over-feeding in this two kinds of geese were researched connected with the earlier period growth performace, the carcass traits,MSTN gene expression,IGF-Ⅰand GH level in serum which closely related with growth development.The results summarized as follows:Experiment 1 Effect of Energy on Male Landes Geese Growth Performance on 0-70 Days StageObjective:To research effect of different ME level on growth performance,the carcass trait,body measurement,serum Parameters,hormone level and muscle composition of male landes geese in pre-overfeeding(0-70 days).Methods:90 0-day-old Landes geese about similar body weight each were used to study the effects of diets containing different metabolic energy levels(0～21d:13.38MJ/kg,12.13MJ/kg and 10.87MJ/k. 22～70d:13.81MJ/kg、12.55MJ/kg and 11.30MJ/kg) through adding soybean oil with equal CP%on bird growth performance,dressed performance,the carcass traits,body measurement,partial serum parameters and IGF-Ⅰ,GH levels in serum.Results:(1) The best ME to male geese is 12.13MJ/kg(0-21d) and 12.55MJ/kg(22-70d) in order to obtaining the best growth performance.(2) Using soybean oil as energy adding,the dressed carcass percentage,eviscerated carcass weight rate,semi-eviscerated carcass weight rate,leg muscle rate of male landes geese were showed no significantly differences(p＞0.05) between these treatments.(3) Protein content in landes male geese’s muscle was all beyond 21%whatever fed with any kinds of energy.Total fattiness content of chest and leg muscle increased with daily food energy concentration,in this experiment that of♂Ⅰwhich food intake energy was the highest respectively higher 24.22%and 34.73%than that of♀Ⅲwhich food intake energy was the lowest;each composition content in chest muscle was larger than that in leg muscle.(4)TG,LDL and HLDL in serum were showed no significantly differences(p＞0.05) between these treatments at 21d; at 70d,the AST,LDL and HLDL in serum were showed no significantly differences (p＞0.05).(5) IGF—Ⅰlevel in serum at 21d was higher than that at 70d except♂Ⅲ.IGF-Ⅰlevel showed significantly differences(p＜0.05) at 21d.Male landes geese’s GH levels in serum at 21d was increased with ME,at 70d,that difference of all groups was unremarkable(p＞0.05).Experiment 2 Effect of Energy on female Landes Geese Growth Performance on 0-70 Days StageObjective:To research effect of different ME levels on growth performance,the carcass trait,body measurement,serum Parameters,hormone level and muscle composition of female landes geese in pre-overfeeding(0-70 days).Methods:According to the design of experiment 1 above.Results:(1) The best ME to female geese is 13.38MJ/kg (0-21d) and 13.81MJ/kg(22-70d) in order to obtaining the best growth performance.(2) Muscle basic composition analysis of 70d female landes geese showed the dry material content of leg muscle was higher than chest muscle about 4.58%,0.21%and 10.29%in the same treatment,total fattiness content of leg muscle was higher than chest muscle about 1.53%,6.91%and 1.88%in the same treatment.(3) Using soybean oil as energy adding,the TG,LDL and HLDL in serum were showed no significantly differences (p＞0.05) between these treatments at 21d.At 70d,the AST,LDL TC AKP and HLDL in serum were showed no significantly differences(p＞0.05).(4) IGF—Ⅰlevel in serum at 21d was higher than that at 70d each treatment.IGF-Ⅰlevel showed no significantly differences(p＞0.05) at 21d,while it showed significantly differences(p＜0.05) at 70d,; female landes geese’s serum GH level at 21d was increased with ME.Experiment 3 Effect of Energy on Xupu Geese Growth Performance on 0-70 Days StageObjective:To research effect of different ME level on growth performance,the carcass trait,body measurement,serum Parameters,hormone level and muscle composition of Xupu geese in pre-overfeeding(0-70 days).Methods:According to the design of experiment 1 above.Results:(1) The best ME to Xupu geese is 13.38MJ/kg (0-21d) and 13.81MJ/kg(22-70d) in order to obtaining the best growth performance.(2) The carcass rate,eviscerated carcass rate,semi- eviscerated carcass rate,abdomen fat ratio and chest muscle rate were significant(p＜0.05) compared to the second group and the other two groups.As for subcutaneous fat layer thickness and intermuscle fat thickness,diversity were significant(p＜0.05) amone three groups.Abdomen fat weight and abdomen fat ratio of the second group were high than other two groups(p＜0.05).(3) Protein content of breast and leg muscle of Xupu geese which intake different energy diety has not significant diversity among three groups.Total lipids of chest muscle of the first group higher 4.46%,8.66%than the second and the third group,respectivelly.Total lipids of leg muscle of the first group higher 4.99%,11.63%than the second and the third group respectivelly.(4)There have no significant differences(p＞0.05) of IGF-Ⅰlevels at 21d or 70d.GH levels in serum of Xupu geese at 21d were low than its at 70d. Conclusions of Experiment 1-3:(1) The best ME to male geese is 12.13MJ/kg(0-21d) and 12.55MJ/kg(22-70d) in order to obtaining the best growth performance.The best ME to female geese is 13.38MJ/kg(0-21d) and 13.81MJ/kg(22-70d) in order to obtaining the best growth performance.The best ME to Xupu geese is 13.38MJ/kg(0-21d) and 13.81MJ/kg(22-70d) in order to obtaining the best growth performance.(2)In this experiment,ME could influence body sizes in limited range;(3) ME influence some parameters which have closely relation with growth performance like slaught weight of male landes geese,eviscerated carcass weight,semi-eviscerated carcass weight,breast muscle weight,leg muscle weight and abdominal fat weight of landes geese were showed significantly differences(p＜0.05) between these treatments;but dressed carcass percentage,eviscerated carcass weight rate,semi-eviscerated carcass weight rate,leg muscle rate of landes geese were showed no significantly differences(p＞0.05) which effected by ME level.(4)ME levels influenced Xupu geese’s dressed carcass percentage, eviscerated carcass weight,semi-eviscerated carcass weight rate,abdominal fat rate and breast muscle rate significantly(p＜0.05).(5)Muscle lipid content of landes geese was higher than it of Xupu geese.CP,total lipid and DM content of male landes geese were higher than those of female landes geese,and the chemical constitution of geese muscles would changed with ME levels.(6) GH levels in serum were influenced by ME levels, while there was a positive relationship between them;IGF-Ⅰlevels in serum has no relationship with ME levels.Experiment 4 The Research of MSTN Gene Clone and energy Influence on MSTN Gene Expression in Landes Geese and Xupu GeeseObjective:Cloning of MSTN gene of Landes geese and Xupu geese the relationship was researched between MSTN gene expression and different energy diets or IGF-Ⅰ,GH in serum in order to know the function of MSTN gene and provide information for molecular nutriology and breeding of waterfowl.Methods:Genome RNA was extracted from muscle of Landes geese and Xupu geese,MSTN gene mRNA was amplified by using RT-PCR,the PCR product was cloned into pGEM-T vector.The MSTN gene was isolated and sequenced from the positive clones screened Based on the MSTN gene clone, usingβ-actin as inner control,constructing an optimal semi-quantitative RT-PCR method, the difference of MSTN gene expression at 21 and 70 days by feeding different energy diets,at one time we detected IGF-Ⅰand GH in serum.Results:(1) The unique DNA fragment about 1128bp in length was obtained.Sequence analysis revealed that it has a length of 1128 nucleotides which encodes a 375 amino peptide.Compared with homologous sequence of human,rat,duck and goose,it displayed a fairly high degree conservation.The homologue of the nucleotide sequence are 94%,94%,99%respectively, the homologue of the amino acid sequence are 98%,97%,98%sespectively.(2) At 21 days,the expressions of MSTN in Landes geese or Xupu geese were showed no significantly differences.(3) At 70 days,the expression of MSTN in male wasⅠ＞Ⅲ＞Ⅱby 70d,to female wasⅢ＞Ⅱ＞Ⅰby 70d,to Xupu geese wasⅢ＞Ⅱ＞Ⅰby 70d.(4) In 21～70 days period,the expression of MSTN was changed with IGF-Ⅰin serum,it was not associated with GH in serum.GH was significant correlations with energy.Conclusions: The different energy influenced MSTN gene expression of landes geese and Xupu geese; in this period,the MSTN gene expression was associated with IGF-Ⅰin serum,but it was not associated with GH in serum.更多还原