【作者】 袁勇超；

【导师】 龚世园；

【作者基本信息】 华中农业大学 ， 水产养殖， 2011， 博士

【摘要】 本研究以胭脂鱼(Myxocyprinus asiaticus)幼鱼为实验对象,研究了胭脂鱼幼鱼饲料中适宜蛋白能量比、磷需求量,胭脂鱼对植物蛋白源的利用及适宜的投喂水平。论文包含6个方面的内容：1.本实验是以体重10g左右的胭脂鱼为研究对象,以蛋白质和能量作为实验因子,设计9种配合饲料,分别饲喂9个组,进行为期8周(56 d)的生长饲养实验,研究适宜于胭脂鱼生长阶段的蛋白质、能量需要量。选取实验鱼共600尾,随机分成9组,每组3个重复,每个重复20尾。实验日粮的营养水平为：蛋白质水平分别为340.0、390.0、440.0 g Kg-1；每个蛋白水平下设置三个脂肪水平分别为60、100和140 g Kg-1,实验饲料的蛋白能量比变化范围为22.4-32.8mg protein kJ-1。实验结果表明饲料蛋白能量比对胭脂鱼的生长性能和体成分有显著影响(P<0.05)。饲料组8和9(蛋白水平为440 gKg-1,可消化能量水平为14.05和15.09 KJg-1,蛋白能量比为31.43和29.22 mg KJ-1)的特定生长率表现出最高(2.16和2.27%d-1)。然而,饲料组6胭脂鱼对饲料利用(蛋白水平为390.0 g Kg-1,可消化能量水平为14.96 KJ g-1)的蛋白质效率、蛋白质积存率和能量积存率表现出最高。实验期间饲料各处理组对胭脂鱼的存活率没有显著影响。极差分析表明实验因子蛋白质水平对胭脂鱼全鱼蛋白含量、胭脂鱼肥满度及肝体指数有显著影响(P<0.05)。实验因子能量对胭脂鱼全鱼、背部白色肌肉和肝脏中粗脂肪含量有显著影响(P<0.05)。基于以上实验结果表明,在本实验条件下,胭脂鱼生长阶段适宜蛋白水平为440.0 g Kg-1,可消化能量水平为14.05-15.09 KJ g-1,适宜蛋白能量比为29.22-31.43 mg KJ-1。饲料中脂肪含量在胭脂鱼生长阶段表现出了一定的蛋白质节约效应。2.本实验拟通过在配合饲料中添加Ca(H2PO4)2,设置不同梯度的有效磷水平,研究胭脂鱼幼鱼对磷的需要量。选择体重为(1.77±0.2 g,平均值±标准偏差)的胭脂鱼450尾进行实验,随机分为5组,每组设3个重复,每个桶(玻璃纤维钢桶)放鱼30尾。在以白鱼粉、豆粕为蛋白源的基础饲料中加入不同浓度的Ca(H2P04)2,基础饲料(饲料1)可利用磷水平为3.1 g Kg-1,配制磷含量(g Kg-1饲料干重)分别为6.2、8.6、10.8、13.2和15.8(可利用磷含量分别为3.1、5.3、7.5、9.6和11.8 g Kg-1)的5种实验饲料。实验周期为8周(56 d),每天饱食投喂两次(早晨8：00和下午5：00),水温变化范围为27.5-30.5℃,溶氧≥6mgl-1。饲养结果表明,当饲料中磷含量为10.8g Kg-1时,胭脂鱼的特定增重率显著高于饲料1和饲料2组(P<0.05),蛋白质效率、饲料系数最低,但与饲料4和饲料5组没有显著性差异。饲料各处理组不同磷水平对胭脂鱼幼鱼摄食量没有显著影响(P>0.05)。饲料中磷利用率与磷水平呈显著性负相关(P<0.05),鱼体成分分析表明其脂肪、灰分、钙、磷、镁、锌、铜含量显著受饲料磷水平的影响(P<0.05),但全鱼鱼体锰的含量和钙磷比处理组之间没有显著性差异。饲料磷水平也显著性影响脊柱骨和鳞片中矿物质的含量,脊椎骨中钙磷比与饲料中磷水平呈显著性的负相关,各处理组之间鳞片中钙磷比没有显著性差异。饲料中磷缺乏症主要表现为较低的生长性能,鱼体组织矿物质含量相对降低和鱼体脂肪含量增加。血清化学成分分析表明饲料中磷水平对血清磷酸酶活性、甘油三酯和总胆固醇水平有一定的影响。对鱼体,脊椎骨和鳞片中磷含量进行线模型回归分析表明,胭脂鱼对饲料中有效磷需要量分别为8.3,8.8,8.6 g Kg-1,对增重率进行折线模型回归分析表明,胭脂鱼饲料适宜有效磷的需要量为7.4 g Kg-1。3.饲料蛋白原料表观消化率数据对于配制精确的饲料配方和降低水产养殖环境污染显得尤为重要。本实验设计为研究胭脂鱼(初始体重18.05±0.27g)对7种饲料原料：白鱼粉、红鱼粉(国产)、发酵豆粕、膨化豆粕、豆粕、棉粕和菜籽粕的干物质、蛋白质、脂肪、总能、磷和氨基酸等的表观消化率,实验饲料为参考饲料和测试饲料(70%参考饲料+30%测试原料),以添加0.5%的三氧化二铬作为外源性指示剂。实验结果表明,白鱼粉的干物质表观消化率最高(74.5%),植物性蛋白原料干物质表观消化率的变化范围为54.3-71.6%。胭脂鱼对白鱼粉、红鱼粉、膨化豆粕和发酵豆粕的蛋白质表观消化率较棉粕和菜粕显著性高(P<0.05),植物性蛋白原料蛋白质表观消化率变化范围为72.8-92.3%,动物性蛋白原料蛋白质表观消化率的变化范围为90.3-91.2%。胭脂鱼利用饲料植物性和动物性蛋白原料中脂肪的表观消化率均较高,变化范围为89.8-94.9%。白鱼粉的能量表观消化率最高,其次是红鱼粉,能量表观消化率最低值为棉粕(P<0.05)。胭脂鱼利用饲料动物性蛋白原料及(发酵豆粕和膨化豆粕)中磷的能力较植物性蛋白原料(豆粕、棉粕和菜粕)要显著性高(P<0.05),动物性和植物性蛋白原料中磷的表观消化率的变化范围分别为58.8-61.4%、25.8-56.0%。胭脂鱼对饲料原料中氨基酸的利用效果同对蛋白质表观消化率的结果相近,只是膨化豆粕除外,胭脂鱼对膨化豆粕中几种氨基酸的利用率较低,可能原因是与膨化豆粕的加工过程中高温处理有关,导致部分蛋白质变性被破坏不能有效消化利用。基于以上研究结果,建议发酵豆粕和膨化豆粕可以考虑作为胭脂鱼饲料中鱼粉的良好替代蛋白源。4.本实验设计为用发酵豆粕部分替代鱼粉对胭脂鱼生长性能和饲料利用率的影响。7组等氮实验饲料配制为用发酵豆粕蛋白分别替代鱼粉蛋白的0、15、25、35、45、55和65%。实验用胭脂鱼初始体重为4.59±0.2 g,每组实验饲料设置3个重复,实验周期为8周。实验结果：饲料中发酵豆粕百分含量超过39.1%(发酵豆粕替代鱼粉蛋白高于45%时),胭脂鱼幼鱼的体增重和特定增长率会显著性下降(P<0.05)。胭脂鱼的生长性能和发酵豆粕替代鱼粉蛋白的比例呈显著性的负相关(P<0.05)。随着发酵豆粕替代鱼粉比例的上升,饲料中赖氨酸和蛋氨酸含量在逐渐降低,可能是造成胭脂鱼幼鱼生长性能下降的主要原因。发酵豆粕替代鱼粉处理对胭脂鱼的摄食水平没有显著性影响。投喂全鱼粉蛋白饲料(D-0)组其胭脂鱼的饵料系数最低,蛋白质效率最高,与D-15、D-25和D-35饲料组没有显著性差异。实验饲料处理组D-0、D-15、D-25和D-35其胭脂鱼对饲料的表观干物质消化率(ADMDs)之间没有显著性差异,变化范围为71.17-72.55%；对饲料蛋白质的表观消化率(ADPs)变化范围为89.07-90.08%,当饲料中发酵豆粕蛋白替代鱼粉蛋白超过45%时(D-45、D-55和D-65处理组),胭脂鱼对饲料的表观干物质消化率(ADMDs)和蛋白质的表观消化率(ADPs)变化范围分别为69.74-70.26%和88.54-88.86%。基于以上实验结果表明发酵豆粕可以作为胭脂鱼配合饲料中鱼粉蛋白的替代蛋白原料,饲料中发酵豆粕蛋白替代鱼粉蛋白的含量可以达35%而对胭脂鱼的生长性能、饵料系数和蛋白质效率没有显著影响。5.本实验设计为在六组实验饲料中添加晶体或包膜赖氨酸和／或蛋氨酸对胭脂鱼生长性能和饲料利用的影响,实验周期为8周。试验用胭脂鱼体重初始值为3.3.g左右,每天进行两次饱食投喂,每组3个重复。其中,五组等氮等能的饲料分别设计为在发酵豆粕型饲料中不添加晶体或包膜的赖氨酸和蛋氨酸(NLM组)、添加晶体赖氨酸和蛋氨酸(LM组)、添加包膜赖氨酸(CL组)、添加包膜蛋氨酸(CM组)和添加包膜赖氨酸和蛋氨酸(CLM组)。对照组实验饲料(FM组)设计为以65%的鱼粉蛋白为动物性蛋白源,不添加包膜或晶体氨基酸。实验结果：胭脂鱼摄食CL组、CM和CLM饲料组较没有添加氨基酸的饲料NLM组来讲,其表现出显著性高的体增重和蛋白质效率(P<0.05)。同时添加包膜赖氨酸和蛋氨酸的饲料CLM组胭脂鱼比饲料LM组生长性能有显著性提高(P<0.05)。实验处理对各试验组胭脂鱼的摄食水平和存活率没有表现出显著性差异(P>0.05)。试验处理组之间胭脂鱼鱼体、肌肉的常规营养组成和氨基酸组成有些许差异,差异不显著。胭脂鱼投喂饲料CLM组相比较饲料NLM组获得了显著性高的肥满度、肝体指数和脏体指数(P<0.05)。实验结果表明胭脂鱼饲料中用发酵豆粕替代一定比例鱼粉,同时添加包膜赖氨酸和蛋氨酸能有效的改善发酵豆粕因其赖氨酸和蛋氨酸含量相对缺乏引起的不良生长问题。6.本试验以11.77g的胭脂鱼为实验对象,拟通过8周的投喂生长实验,研究不同摄食水平对胭脂鱼幼鱼生长性能,饲料利用,营养存留,鱼体组成及营养物质的表观消化率的影响。实验周期8周,摄食水平设计为每天投喂量为其鱼体体重的0%(饥饿组)-4%(bw d-1)(递增水平为0.5%),共9个水平,每个水平设置3个重复。实验结果表明,不同摄食水平对胭脂鱼的生长性能,饲料利用率,营养积存,体组成和对饲料干物质、蛋白质和能量的表观消化率都有显著性影响(P<0.05)。饥饿处理组胭脂鱼幼鱼存活率最低。胭脂鱼实验末鱼体体重,生长率,随摄食水平递增至3.0%而显著性上升(P<0.05)。饲料系数在摄食水平为2.5%时表现出最低(P<0.05)。蛋白质效率随摄食水平递增至2.5%而显著性上升(P<0.05)。当摄食水平为1.0%(每尾鱼每天摄入能量≤2.27 kJ)时,鱼体维持基本正常代谢,表现出了摄食水平时蛋白质积存效应优先于脂肪积存。这表明当鱼体摄食水平处于新陈代谢维持水平时,鱼体内脂肪组织会支持蛋白质优先积存。全鱼,肌肉和肝脏粗脂肪含量,全鱼蛋白质含量随投喂水平从0.5%递增至3.0%而显著性上升(P<0.05),然而摄食水平对肌肉和肝脏蛋白质含量没有显著性影响(P>0.05)。胭脂鱼幼鱼对饲料中干物质、蛋白质和能量的表观消化率随摄食水平递增(0.5-3.0%)而显著性上升。对胭脂鱼幼鱼生长率进行折线模型回归分析表明胭脂鱼幼鱼适宜生长性能的摄食水平和维持体代谢水平分别为3.10%和0.45%。更多还原

【Abstract】 Optimum protein and energy levels, feeding levels and phosphorus requirements and utilization of plant protein source of juvenile Chinese sucker were determined in the present study. The results obtained can be briefly summarized as follows:1. A growth experiment was conducted to investigate effect of dietary protein to energy ratios on growth and body composition of juvenile Myxocyprinus asiaticus (initial mean weight:10.04±0.53 g, mean±S.D.). Nine practical diets were formulated to contain three protein levels (340,390 and 440 g Kg-1), each with three lipid levels (60, 100 and 140 g Kg-1), in order to produce a range of P/E ratios (from 22.4 to 32.8 mg protein kJ-1). Each diet was randomly assigned to triplicate groups of 20 fish in 400-L indoors flow-through circular fibreglass tanks provided with sand-filtered aerated freshwater. The results showed that the growth was significantly affected by dietary P/E ratio (P<0.05). Fish fed the diets with 440 g Kg-1 protein (100 and 140 g Kg-1 lipid, P/E ratio of 31.43 and 29.22 mg protein kJ-1) had the highest specific growth rates (SGR) (2.16 and 2.27% day-1, respectively). However, fish fed the diet with 390 g Kg-1 protein and 140 g Kg-1 lipid showed comparable growth (2.01% day-1), and had higher protein efficiency ratio (PER), protein productive value (PPV) and energy retention (ER) than other groups (P<0.05). No significant differences in survival were found among dietary treatments. Carcass lipid content was positively correlated with dietary lipid level, but irrespective of protein level and inversely correlated with carcass moisture content. Carcass protein contents increased with increasing dietary lipid at each protein level. The white muscle and liver composition showed that lipid increased with increasing dietary lipid level (P<0.05). Dietary protein concentrations had significant effect on condition factor (CF), hepatosomatic index (HSI) and viscerosomatic index (VSI) (P<0.05). However, Dietary lipid concentrations had no significant effect on CF, HSI (P>0.05). Based on these observations,440 g Kg-1 protein with lipid from 100 to 140 g Kg-1 (P/E ratio of 29.22 to 31.43 mg protein kJ-1) seemed to meet minimum requirement for optimal growth and feed utilization, and lipid could cause protein-sparing effect in diets for juvenile Chinese sucker.2. A growth trial was conducted to estimate the optimum requirement of dietary available phosphorus (P) for Chinese sucker juveniles. Triplicate groups of juveniles Chinese sucker (initial mean weight:1.77±0.02 g, mean±S.D.) were fed diets containing graded levels (3.1,5.3,7.5,9.6 and 11.8 g kg-1) of available phosphorus. The basal diet (diet 1), containing 3.1 g kg-1 available P, was supplemented with graded levels of monocalcium phosphate (MCP) to formulate four experimental diets. The fish were fed twice daily (08:00 h and 17:00 h) to satiation for 8 weeks. During the experimental period, the water temperature fluctuated from 27.5 to 30.5℃and dissolved oxygen was more than 6 mg l-1. The specific growth rate (SGR), protein efficiency ratio (PER) were all significantly increased by dietary available phosphorus up to 7.5 g kg-1 (P<0.05) and then leveled off beyond this level. Feed conversion ratio (FCR) significantly decreased with dietary available phosphorus level up to 7.5 g kg-1 (P<0.05). Dietary treatments did not significantly affect feed intake (FI) (P>0.05). Efficiency of phosphorus (P) utilization significantly decreased with dietary available phosphorus level (P<0.05). Body composition analysis showed that the whole-body lipid, ash, calcium, phosphorus, magnesium(Mg), zinc(Zn), copper(Cu) contents were all significantly affected by dietary available P concentration (P<0.05), however, no significance were found Manganese(Mn) concentration and calcium/phosphorus (Ca/P) ratios in whole-body among all the treatments (P>0.05). Dietary phosphorus levels also significantly affected the mineralization of vertebrae and scale (P<0.05), and Ca/P ratios in scale were not influenced by dietary P supplementation, while vertebrae Ca/P ratio decreased with dietary available P levels (P<0.05) (quadratic effect, P<0.001). Signs of phosphorus deficiency were characterized by poor growth, slightly reduced mineralization and an increase in body lipid content. The blood chemistry analysis showed that dietary available P had distinct effects on enzyme activities of alkaline phosphatase (ALP), as well as contents of triacyglycerol (TG) and total cholesterol (T-CHO) (P<0.05). Broken-line analysis based on weight gain (WG) indicated the minimum available phosphorus requirement for the optimal growth of juvenile Chinese sucker was 7.4 g kg-1. Based on the phosphorus content in whole body, vertebrae or scale indicated that the requirements were 8.3,8.8 and 8.6 g kg-1, respectively.3. Building a reliable database on apparent digestibility of protein sources is critical for accurate feed formulation and reduction of aquaculture wastes. Apparent digestibility coefficients (ADCs) of dry matter (DM), crude protein, crude lipid, gross energy, phosphorus and amino acids for white fish meal (WFM), brown fish meal (native) (BFM), fermented soybean meal (FSBM), extruded soybean meal (ESBM), soybean meal (SBM), cottonseed meal (CM) and rapeseed meal (RM) were determined for Chinese sucker (Myxocyprinus asiaticus) (18.05±0.27g) using a reference diet (RF) and test diets (70% RF diet plus 30% of tested feedstuff) containing 0.5% chromic oxide as an inert marker. WFM had the highest apparent digestibility coefficient for DM (74.5%). For plant products, the ADCs of DM ranged from 54.3 to 71.6%. Protein digestion by Chinese sucker of WFM, BFM, FSBM and ESBM was significantly higher than CM and RM, ranging from 72.8 to 92.3% for plant products, and 90.3 to 91.2% for animal feedstuffs. Lipids from both animal and plant feedstuffs were highly digested by Chinese sucker, ranging from 89.8 to 94.9%. The highest digestibility coefficient of energy was registered for WFM, followed by BFM; and the lowest digestibility coefficient of energy was recorded for CM (P<0.05). The Chinese sucker did utilize dietary phosphorus from the animal feedstuffs (FSBM and ESBM) more efficiently than from plant feedstuffs (SBM, CM and RM), with values ranging from 58.8 to 61.4% and 25.8-56.0%, respectively. Amino acid availability reflected protein digestibility, except for ESBM, for which the availability of some amino acids was lower, possibly because of protein denaturation during processing. Based on the current findings, FSBM and ESBM could be considered as good potential substitutes for FM in diets for Chinese sucker.4. The study was designed to study the growth performance of the Chinese sucker, Myxocyprinus asiaticus in response to the replacement of fish meal with Fermented Soybean meal (FSBM) in its diet. Seven isonitrogenous and isolipidic diets were formulated with 0,15,25,35,45,55 or 65% replacement of fish meal with FSBM on a protein basis. Each diet was fed to three replicate groups of fish with an initial weight 4.59±0.2 g for eight weeks. Weight growth (WG) and Specific growth rate (SGR) was significantly lower when FSBM inclusion was 45% or greater, replacing more than 45% fish meal protein. A significant negative relationship was observed between growth response and the level of fish meal protein replacement with FSBM protein. Methionine and Lysine content decreased as FSBM inclusion levels increased, consequently compromising growth performance. Feed intake (FI) were unaffected by fish meal replacement levels. The feed conversion ratio (FCR) and the protein efficiency ratio (PER) were highest at the lowest FSBM inclusion level. Experimental diets D-0, D-15, D-25 and D-35 had apparent dry matter digestibility (ADMDs) ranging from 71.17-72.55% and apparent protein digestibility (ADPs) from 89.07-90.08%, while the diets with higher FSBM inclusion (D-45 to D-65) had a significantly lower ADMDs range (69.74-70.26%) and APDs range (88.54-88.86%). It is concluded that FSBM is an acceptable alternative plant protein source that can replace up to 35% of fish meal protein in diets without significant adverse effects on growth, survival, FCR, PER and body composition.5. An 8-week feeding trial was conducted using six test diets to evaluate the effects of supplementation of crystalline or coated lysine (Lys) and/or methionine (Met) on growth performance and feed utilization of the Chinese sucker, Myxocyprinus asiaticus. Test diets were fed to triplicate groups of Chinese sucker (about 3.3 g) two times a day. Five iso-nitrogenous and iso-caloric diets based on fermented soybean meanl (FSBM) were prepared by either not supplementing with lysine and methionine (NLM), or supplementing with both crystalline Lys and Met (LM), with coated Lys (CL), with coated Met (CM), or with both coated Lys and Met (CLM). The control diet (FM) was formulated to contain 65% fish meal as the sole animal protein source without amino acid (AA) supplementation. The Chinese sucker juveniles fed the CL, CM and CLM diets had higher weight gain (WG) and protein efficiency ratio (PER) compared to Chinese sucker fed the NLM diet (P<0.05). The Chinese sucker juveniles fed the CLM diet showed significantly higher growth performance than those fed the LM diet (P< 0.05). Feed intake (FI) and survival among all groups at the end of the experiment were not significantly affected by the dietary treatments. The whole body, muscle proximate, total amino acid composition showed few differences among the groups. The Chinese sucker juveniles fed the CLM diet showed significantly higher CF, HSI and VSI than those fed the NLM diet. This study indicated that the supplementation of coated lysine and methionine was effective in improving the nutritive value of FSBM-based diets deficient in methionine and lysine for juvenile Chinese sucker.6. An experiment was conducted to determine effects of feeding levels on growth performance, feed utilization, nutrient deposition, body composition, and apparent digestibility coefficients (ADCs) of nutrients for juvenile Chinese sucker (initial weight, 11.77±0.22 g). Chinese sucker were fed a practical diet from 0 (starvation) to 4.0% (at 0.5% increments) body weight per day (bw d-1) for 8 weeks. The results showed that growth performance, feed utilization, nutrient deposition, body composition, and ADCs of dry matter, protein, and energy were significantly (P<0.05) affected by feeding levels. Survival was the lowest for the starvation group. Final mean body weight, Growth rate, thermal-unit growth coefficient (TGC) increased with feeding rate from 0 to 3.0% bw d-1 (P<0.05) and showed no significant differences above the level (P>0.05). Feed conversion rate was significantly lower at a feeding level of 2.5% bw d-1 than above and below the level (P<0.05). Protein efficiency ratio was markedly highest at the 2.5% bw d"1 ration level (P<0.05). Fish fed at the feeding level (1.0% bw d-1), which represented a maintenance ration (energy gain was less than 2.27 kJ fish-1 day-1), showed positive protein deposition but negative lipid deposition. This indicates that fish fed a maintenance ration mobilize body lipid reserve to support protein deposition. Lipid contents of whole body, muscle, and liver increased with increasing feeding rates from 0.5 to 3.0% bw d-1 and showed no significant differences above the level (P>0.05). Protein contents of whole-body composition increased with feeding rate from 0.5 to 3.0% bw/d (P<0.05) and showed no significant differences above the level (P>0.05), whereas muscle, and liver remained relatively stable with the different ration amount with the exception of fish fed a ration of 0.5% bw d-1, at which Chinese sucker possessed significantly lower body protein concentration (P<0.05). ADCs of dry matter, protein, and energy decreased with increasing feeding levels from 0.5 to 3.0% bw d-1 and then remained relatively constant over the level. Based on the broken-line regression analysis using WG data, the optimum and maintenance feeding levels for Chinese sucker were 3.10% bw d-1 and 0.45% bw d-1, respectively更多还原