CN112273563A - 一种适用于高植物蛋白源的鲟鱼配合饲料的添加剂及应用 - Google Patents
一种适用于高植物蛋白源的鲟鱼配合饲料的添加剂及应用 Download PDFInfo
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- CN112273563A CN112273563A CN202011362561.8A CN202011362561A CN112273563A CN 112273563 A CN112273563 A CN 112273563A CN 202011362561 A CN202011362561 A CN 202011362561A CN 112273563 A CN112273563 A CN 112273563A
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Abstract
本发明涉及饲料添加剂领域,提供了一种适用于高植物蛋白源的鲟鱼配合饲料的添加剂及应用。本发明的添加剂由牛磺酸和硫辛酸构成。本发明在当前鲟鱼饲料中大量使用植物蛋白原料,造成鲟鱼生长速度降低、饲料系数升高、氧化应激增加的背景下,提供了一种组方简单,在高植物蛋白源配合饲料中添加后可明显提高鲟鱼生长性能、改善饲料利用高效率、降低氧化应激水平的饲料添加剂。
Description
技术领域
本发明涉及饲料添加剂领域,特别是涉及一种适用于高植物蛋白源鲟鱼配合饲料的添加剂及应用。
背景技术
鱼粉具有较高的营养价值、适口性和消化率,是水产饲料的首选蛋白源。可是,近年来随需求增加而资源受限等问题的出现,鱼粉价格持续处于高位且波动较大,所以需要寻找合适的替代蛋白原料。植物蛋白原料因价格低廉且供应稳定,是理想的替代原料。但大多数研究发现,一方面大多植物蛋白原料中营养不平衡,且含有毒有害物质或抗营养因子等因素,导致鱼类生长与健康受到负面影响;另一方面,由于高植物蛋白原料带来饲料可消化糖类的增加,导致鱼体脂肪沉积异常,引起氧化应激,造成代谢负担。这些问题在幼鱼阶段显得尤为突出,而幼鱼阶段的生理状况与商品鱼养成阶段的生长表现呈正相关。因此,如何减轻甚至消除高植物蛋白替代鱼粉所引发的影响,对水产饲料的生产有着重要的意义。
目前,以提高植物蛋白源的营养价值并减轻其负面影响为目的的技术方案大致可分为三类,即混合植物蛋白源、加工处理原料(如膨化、发酵等)以及使用饲料添加剂。其中,饲料添加剂因具品种多、作用各异、质量相对稳定的特点,更是这些技术方案中的研究热点。
鲟鱼,即鲟形目鱼类,部分种兼具海淡水生活史。世界鲟鱼的规模化养殖起步于上世纪八十年代,目前中国是鲟鱼养殖产量最大国,约占世界养殖总产量的八成。近年来,随着养殖技术的成熟,商品鲟鱼每公斤的塘边销售价格从高峰的百元跌至目前二十元左右。饲料成本占水产动物养殖成本的60%左右。因此,为降低养殖成本获取养殖效益,对饲料成本提出了很大要求。这就需要对以鱼粉等高价格原料为主的饲料配方进行优化升级,以降低饲料成本。大部分鲟鱼为偏肉食性鱼类,在一定程度上可以利用植物蛋白,然而饲料中使用高比例植物蛋白原料替代鱼粉,鲟鱼的生长或饲料利用会受到抑制,肠道和肝脏功能受损,鱼体尤其是肝脏脂肪沉积增加。因此,有必要开发适合鲟鱼高植物蛋白源饲料配方的添加剂。
牛磺酸是一种含硫的类氨基酸,被认为是鱼粉中的活性小分子之一,具有调节诸如运动行为、食物消耗、能量代谢和胆汁酸合成等作用。除了猫科等少数几种哺乳动物不能合成牛磺酸外,大多数脊椎动物具有一定的牛磺酸合成能力,但在幼龄期体内牛磺酸合成量相对不足,满足不了生长发育的需求,需要外源补充。而植物原料普遍缺乏牛磺酸,这预示着高植物蛋白源配合饲料中牛磺酸的含量可能不足。多数研究发现饲料中添加适量的牛磺酸有利于鱼类生长,提高抗氧化能力,改善鱼类健康,海水鱼类的添加适宜水平似乎高于淡水鱼类。而投喂缺乏牛磺酸的饲料确实会造成鱼类生长减缓、疾病易感性增加,仔稚鱼发育受阻,这在海水鱼类中尤为明显;同时,也发现摄入缺乏牛磺酸的饲料,会导致有些鱼还出现绿肝综合征(green liver syndrome),而补充牛磺酸后鱼类生长性能得到改善,绿肝综合征的发生率也大为降低。但也有少数的研究发现,牛磺酸的添加量过多,会给鱼类健康带来负面影响。已有研究发现,在高植物蛋白源饲料中添加适量的牛磺酸(饲料中添加1000mg/kg牛磺酸)后可促进鲟鱼生长,但会引起脂肪代谢发生改变,造成血脂(胆固醇和甘油三酯)水平升高;然而,高水平的添加(饲料中添加大于2500mg/kg牛磺酸)后反而会抑制其生长,并对肝脏健康不利,表明高植物蛋白饲料中仅添加牛磺酸不足以解决高植物蛋白源饲料对鲟鱼所产生负面问题。因此,亟需研发适合高植物蛋白源饲料配方的复合添加剂。
硫辛酸是一种类维生素的含硫有机羧酸,具有金属鳌合能力、清除活性氧、再生内源性抗氧化剂和修复氧化损伤的作用。硫辛酸的结构特征,使其易被吸收且能分布到机体的各部位发挥作用,甚至能透过血脑屏障,被认为是一种“理想的抗氧化剂”,保护机体免受氧化损伤。而进一步发现饲料中添加硫辛酸可调节鱼类脂肪代谢,促进体内脂解及脂肪酸β-氧化,改善饲料蛋白质利用率与体蛋白沉积率。因此,在高植物蛋白源饲料中同时添加牛磺酸和硫辛酸是否会提高鲟鱼对高植物蛋白源饲料的利用,是否存在协同作用,改善鱼体健康,现在尚未有报道发表。
发明内容
本发明的目的是针对当前鲟鱼配合饲料中植物蛋白源的比例使用较高,导致鲟鱼生长速度减缓、氧化应激增加和健康受损的背景下,提供一种适用于高植物蛋白源的鲟鱼配合饲料的添加剂。
本发明的另一个目的在于提供一种适用于高植物蛋白源的鲟鱼配合饲料的添加剂的应用。
为实现上述目的,本发明通过以下技术方案实现:
一种适用于高植物蛋白源的鲟鱼配合饲料的添加剂,由以下组分按重量份组成:牛磺酸50-100份,硫辛酸30-60份,米糠粕840-920份;
以上所述的方案中,优选的,牛磺酸50-80份,硫辛酸30-45份,米糠粕880-920份。
一种适用于高植物蛋白源的鲟鱼配合饲料的添加剂的应用,包括利用该添加剂制备成高植物蛋白原的鲟鱼饲料;
以上所述的应用中,所述的鲟鱼饲料是由下述组分按重量份组成:
所述的植物蛋白1为大豆分离蛋白或膨化大豆。
以上所述的应用中,优选的,所述的饲料可促进鲟鱼生长,提高鱼体蛋白合成和脂肪代谢,增强鲟鱼免疫力。
以上所述的应用中,优选的,所述的鲟鱼为杂交鲟。
与现有技术相比,本发明具有以下优点:
本发明提供了一种适用于高植物蛋白源鲟鱼配合饲料的添加剂及应用。与现有鲟鱼饲料添加剂相关的技术先比,一方面,本发明是特别适用于高植物园的鲟鱼饲料的添加剂;另一方面,本发明的添加剂组方简单,二种主要成分配伍具有协同作用。在高植物蛋白源饲料中使用本发明的添加剂,可提高鲟鱼对高植物蛋白源配合饲料的利用,促进生长,提高鲟鱼的抗氧化能力,降低病害的发生。
具体实施方式
下面结合具体实施例对本发明做进一步说明,但本发明不受实施例的限制。本发明所述技术方案,如未特别说明,均为本领域的常规方案;所述试剂或材料,如未特别说明,均来源于商业渠道。本发明实施例所用牛磺酸的纯度≥99%,硫辛酸的纯度≥99%。
实施例1:
一种适用于高植物蛋白源的鲟鱼配合饲料添加剂,按以下重量份的原料制成:牛磺酸80份,硫辛酸40份,米糠粕880份,混合粉碎后过60目筛即得。
实施例2:
一种适用于高植物蛋白源的鲟鱼配合饲料添加剂,按以下重量份的原料制成:牛磺酸50份,硫辛酸30份,米糠粕920份,混合粉碎后过60目筛即得。
实施例3:
一种适用于高植物蛋白源的鲟鱼配合饲料添加剂,按以下重量份的原料制成:牛磺酸70份,硫辛酸45份,米糠粕885份,混合粉碎后过60目筛即得。
实施例4:
鲟鱼配合饲料添加剂在制备高植物蛋白由的鲟鱼饲料中的应用:
1.实验鱼
来源于专业鲟鱼繁育场,为国内最主要的养殖鲟鱼杂交鲟(Acipenser baerii♀×A.schrenckii♂)。运回后,在养殖系统中暂养2周。
2实验饲料
以鱼粉为动物蛋白源,以豆粕、大豆分离蛋白、菜粕、花生粕、玉米蛋白粉为植物蛋白源,配制成以鱼粉为主要蛋白源的正对照组饲料(P),以大豆分离蛋白、豆粕、菜粕、花生粕和玉米蛋白粉等植物蛋白源为主的负对照组饲料(N)。另外,在负对照组饲料配方及基础上分别以牛磺酸(T8)、硫辛酸(L4)以及实施例1(T8L4)作为添加剂,配制成2组对比饲料和1种实施例饲料,共计5种饲料。详细试验饲料配方和营养成分见表1。
表1饲料配方和营养成分(g/kg)
3养殖实验与管理
正式实验前用正常基础饲料暂养2周,使实验鱼适应养殖系统和实验饲料。选取体质健壮、规格整齐的初始体质量为29.36±0.07g的实验鱼225尾,放入1套流水养殖系统的18个养殖桶(有效容积450L)中,每桶放鱼15尾。将桶随机分为5组,每组3个平行,每天投喂三次,表观饱食投喂。投喂时间分别为8:30、12:30和16:30,每天记录投喂量。养殖周期42d。养殖期间水温保持在19℃-21℃,溶氧大于5mg/L。
4样品采集及检测方法
饲养28d后,于每晚20时收集粪便,冷冻干燥后放入-20℃冰箱,以备测定干物质、能量、蛋白质、脂肪等的表观消化率。
养殖实验结束后,禁食24h,麻醉(MS-222,100mg/L)后,对每桶实验鱼进行记数和称体重,用于计算成活率、增重率,并根据饲料投喂量计算饲料效率、蛋白质效率。每桶随机取鱼3尾,测量体长和体重,用于计算肥满度;随后取内脏称重,分离肝脏并称重,肠道长度,用于计算肝体比、脏体比、肥满度。并分离并保留背肌、肝脏、用于测定常规营养成分。并另取3尾,用于测定全鱼常规营养成分。营养成分的测定均采用相关的中华人民共和国国家标准。
养殖实验结束后每桶随机取鱼3尾,于尾静脉取血,4℃冰箱内静置2h,以3000r/min的转速离心15min,得到空腹血清,保存于-20℃冰箱中,用于测定血清生化指标和抗氧化指标。分离肝脏,取相同部位的于液氮速冻后,保存于-80℃冰箱中,用于测定肝脏生化指标。其余各指标的计算公式如下:
增重率(WG,%)=(FW-IW)×100/IW
特定生长率(SGR,%/d)=(lnFW-lnIW)×100/t
饲料系数(FCR)=WA/(Wf-Wi+Wd)
成活率(SR,%)=Nf×100/Ni
肥满度(CF)=W×100/L3
肝体比(LSI,%)=WL×100/W
脏体比(VSI,%)=WV×100/W
蛋白质沉积率(PRE,%)=[WA×Wp-(Wf-Wi+Wd)×Fp]×100/(WA×Wp)
干物质表观消化率(ADCd,%)=100×(1-Nf/Mf)
营养物质表观消化率(ADCe,%)=100×(1-Nf/Nd×Md/Mf)
其中,t为实验天数(d),FW和IW分别为每周期实验鱼的初、末均体质量(g),Wi、Wf和Wd分别为每周期实验鱼的初、末总体质量和死亡鱼体质量(g),W为鱼体质量(g),L为鱼体长(cm),Ni和Nf分别为每周期结束后和开始时鱼的尾数;WL、WV分别为肝脏质量(g)、内脏团质量(g),WA、Wp和Fp分别为投喂饲料总质量(g),饲料粗蛋白含量(%)和全鱼蛋白含量(%),Md为饲料中标记物含量(%),Mf为粪便中标记物含量(%),Nd为饲料中营养成分的含量(%),Nf为粪便中营养成分的含量(%)。
5.统计分析
采用SPSS18.0对所得数据进行方差分析(one-way ANOVA),当差异显著时(P<0.05),采用Tukey检验进行多重比较。
6.结果
6.1生长性能
由表2可见,经过6周的养殖,各处理组间成活率较高,且无显著差异(P>0.05)。但是,正对照组的末均体质量、增重率特定生长率显著高于N、T8和L4组的(P<0.05),然而实施例1添加组(T8L4)与正对照均无显著差异(P>0.05)。表明通过添加本发明的添加剂,鲟鱼即使摄食在高植物蛋白源的饲料仍可以达到与摄食以鱼粉为主的饲料的水平,体现了本发明在促进鲟鱼生长方面的优点。同时,与正、负对照组相比,本发明的添加剂未对实验鱼的肥满度、肝体比和脏体比产生显著的影响(P>0.05)。
表2生长性能
注:表格中同列相同小写字母或无字母表示差异不显著(P>0.05),不同小写字母表示差异显著(P<0.05)。
6.2饲料利用
由表3可见,实施例1添加组的实验鱼的摄食率、饲料系数与P组无显著差异(P>0.05),但显著低于N和T8组的(P<0.05);而实施例添加组的1实验鱼的蛋白质效率和蛋白质保留率亦与P组无显著差异(P>0.05),但显著高于N和T8组的(P<0.05);L4组的这4个指标均与N组无显著差异(P>0.05),表明实施例1添加组可以改善饲料利用。实施例1添加组的干物质、粗蛋白和能量的表观消化率与L4组差异不显著(P>0.05),但显著高于N组(P<0.05),表明实施例1添加组改善饲料利用的原因是通过提高饲料营养物质的表观消化率实现的。
表3饲料利用
注:表格中同列相同小写字母或无字母表示差异不显著(P>0.05),不同小写字母表示差异显著(P<0.05)。
6.3体成分
由表4可见,各处理组间全鱼和肝脏的营养成分均无显著差异(P>0.05)。但是,与P组相比,实施例1添加组的肌肉水分显著降低(P<0.05),肌肉粗脂肪含量显著升高(P<0.05),表明实施例1添加组的饲料更有利于肌肉脂肪沉积。
表4体成分(%,湿重)
注:表格中同列相同小写字母或无字母表示差异不显著(P>0.05),不同小写字母表示差异显著(P<0.05)。
6.4血清抗氧化指标
由表5可见,各组间过氧化氢酶(CAT)、总抗氧化能力(TAOC)和铜锌超氧化物歧化酶(CUZNSOD)活性没有显著差异,但与P和N组相比,实施例1可以显著降低血清丙二醛(MDA)水平(P<0.05);而与N组相比,实施例1添加组也显著增加了总谷胱甘肽(TGSH)和氧化型谷胱甘肽(GSSG)的含量(P<0.05),且显著增高了溶菌酶的活性(P<0.05),表明实施例1添加组饲料更有利实验鱼健康。与对比例相比,实施例1添加组的鱼体的血清MDA含量显著降低(P<0.05),表明实施例1添加组饲料更能组织脂肪氧化。
表5血清抗氧化指标/(U/L)
注:表格中同列相同小写字母或无字母表示差异不显著(P>0.05),不同小写字母表示差异显著(P<0.05)。MDA丙二醛,CAT过氧化氢酶,TAOC总抗氧化能力,TGSH总谷胱甘肽,GSSG氧化型谷胱甘肽,GSH还原型谷胱甘肽,TSOD总超氧化物歧化酶,CUZNSOD铜锌超氧化物歧化酶,LZM溶菌酶。
6.5血清生化指标
由表6可见,N组血清谷丙转氨酶(AST)显著高于P组(P<0.05),表明高植物蛋白源饲料可能会对肝细胞损伤,而饲料中添加牛磺酸、硫辛酸后可以降低转氨酶(ALT、AST)活性,改善肝脏健康状况。与N组相比,虽然血清总蛋白(TP)、低密度脂蛋白胆固醇(LDLC)水平无显著差异(P>0.05),但是与P组相比,实施例1添加组的可以显著提高血清TP和LDLC水平(P<0.05),表明实施例1添加组可以提高蛋白质合成和脂肪代谢。
表6血清生化指标
注:表格中同列相同小写字母或无字母表示差异不显著(P>0.05),不同小写字母表示差异显著(P<0.05)。ALT谷草转氨酶,AST谷丙转氨酶,ALP碱性磷酸酶,TP总蛋白,ALB白蛋白,TC总胆固醇,
LDLC低密度脂蛋白胆固醇,HDLC高密度脂蛋白胆固醇,TG甘油三酯,GLU血糖。
6.6肝脏抗氧化
由表7可见,N组的MDA、TGSH、GSH含量,CAT、TSOD、CUZNSOD活性均高于P组(P<0.05),这表明N组的氧化应激状态高于P组,而饲料中添加牛磺酸、硫辛酸后可以缓解应激状态,表现为MDA、TGSH和GSH水平下降,CAT、TSOD、CUZNSOD活性降低。但实施例1添加组与对比例相比,更有利于肝脏CAT活性(P<0.05)。
表7肝脏抗氧化/(U/mg protein)
注:表格中同列相同小写字母或无字母表示差异不显著(P>0.05),不同小写字母表示差异显著(P<0.05)。MDA丙二醛,CAT过氧化氢酶,TAOC总抗氧化能力,TGSH总谷胱甘肽,GSSG氧化型谷胱甘肽,GSH,还原型谷胱甘肽,TSOD,总超氧化物歧化酶,CUZNSOD,铜锌超氧化物歧化酶,LZM溶菌酶。
实施例5:
1.实验鱼
来源同实施例4。
2实验饲料
以鱼粉、鸡肉粉为动物蛋白源,以豆粕、膨化大豆、菜粕、花生粕、玉米蛋白粉为植物蛋白源,配制成以鱼粉为主要蛋白源的正对照组饲料,以大豆分离蛋白、豆粕、菜粕、花生粕和玉米蛋白粉等植物蛋白源为主的负对照组饲料(N);另外,在负对照组饲料配方及基础上分别以实施例2、实施例3作为添加剂,配制成2组实施例饲料。共计4种饲料。详细实验饲料配方和营养成分见表8。
表8饲料配方和营养成分(g/kg)
3养殖实验与管理
正式实验前用正常基础饲料暂养2周,使实验鱼适应养殖系统和实验饲料。选取体质健壮、规格整齐的初始体质量为12.02±0.17g的实验鱼180尾,放入1套流水养殖系统的12个养殖桶(有效容积450L)中,每桶放鱼15尾。将桶随机分为4组,每组3个平行,每天投喂三次,表观饱食投喂。投喂时间分别为8:30、12:30和16:30,每天记录投喂量。养殖周期28天。养殖期间水温保持在19℃-21℃,溶氧大于5mg/L。
4样品采集及检测方法
养殖实验结束后,禁食24h,麻醉(MS-222,100mg/L)后,对每桶实验鱼进行记数和称体重,用于计算成活率、增重率,并根据饲料投喂量计算饲料系数。每桶随机取鱼3尾,测量体长和体重,用于计算肥满度;随后于尾静脉取血,4℃冰箱内静置2h,以3000r/min的转速离心15min,得到空腹血清,保存于-20℃冰箱中,用于测定血清指标。各指标的计算公式见实验例1。
5统计分析
采用SPSS18.0对所得数据进行方差分析(one-way ANOVA),当差异显著时(P<0.05),采用Tukey检验进行多重比较。
6结果
6.1生长性能和饲料利用
经过28天的养殖,实验期间未发现鱼死亡,成活率均为100%。由表9可见,负对照组的实验鱼生长最差,末体质量显著低于其他三组(P<0.05)。相比于负对照组,实施例2添加组和实施例3添加组的末体质量和增重率均有所升高,尤其是实施例3的与正对照组无显著差异(P>0.05),表明实施例添加组可以增加实验鱼的生长速度。
表9生长性能和饲料利用
组别 | 初均体重/g | 末均体重/g | 增重率/% | 肥满度/(g/cm<sup>3</sup>) | 饲料系数 |
正对照组P | 11.82±0.24 | 55.56±1.13<sup>c</sup> | 370.23±12.48<sup>c</sup> | 0.65±0.02 | 0.82±0.02<sup>a</sup> |
负对照组N | 12.07±0.21 | 40.10±1.26<sup>a</sup> | 232.39±9.20<sup>a</sup> | 0.69±0.02 | 1.10±0.03<sup>b</sup> |
实施例2 | 12.53±0.58 | 48.31±0.88<sup>b</sup> | 286.79±16.10<sup>ab</sup> | 0.69±0.03 | 0.88±0.04<sup>a</sup> |
实施例3 | 11.64±0.10 | 49.70±1.98<sup>bc</sup> | 326.85±17.59<sup>bc</sup> | 0.68±0.03 | 0.94±0.04<sup>a</sup> |
注:表格中同列相同小写字母或无字母表示差异不显著(P>0.05),不同小写字母表示差异显著(P<0.05)。
6.2血清抗氧化和免疫指标
由表10可见,与正对照组相比,实施例2添加组无显著差异(P>0.05),但实施例3添加组的溶菌酶活性显著升高(P<0.05),表明实施例似乎提高了实验鱼的免疫力;而GSH-PX和SOD酶活性无显著差异(P>0.05),且这三组的均显著高于负对照组(P<0.05),表明实施例添加组可以提高实验鱼的抗氧化能力。
表10血清抗氧化和免疫指标(U/L)
组别 | 溶菌酶 | GSH-PX | SOD |
正对照组P | 13.68±0.76<sup>b</sup> | 174.42±2.17<sup>b</sup> | 1143.90±7.81<sup>b</sup> |
负对照组N | 9.51±0.83<sup>a</sup> | 149.46±7.84<sup>a</sup> | 894.04±41.32<sup>a</sup> |
实施例2 | 17.53±0.93<sup>c</sup> | 177.94±1.28<sup>b</sup> | 1073.63±49.84<sup>b</sup> |
实施例3 | 13.50±0.63<sup>b</sup> | 180.27±0.82<sup>b</sup> | 1175.43±16.78<sup>b</sup> |
注:表格中同列相同小写字母或无字母表示差异不显著(P>0.05),不同小写字母表示差异显著(P<0.05)。GSH-PX谷胱甘肽过氧化物酶,SOD超氧化物歧化酶。
综合以上结果,可以发现在高植物蛋白源饲料中,添加实施例的添加剂,可以提高鲟鱼的生长速度,改善饲料利用效率,并提高了抗氧化能力,尤其是可以改善因为高植物蛋白源产生的血清抗氧化能力下降问题。
Claims (6)
1.一种适用于高植物蛋白源的鲟鱼配合饲料的添加剂,由以下组分按重量份组成:牛磺酸50-100份,硫辛酸30-60份,米糠粕840-920份。
2.根据权利要求1所述添加剂,其特征在于:牛磺酸50-80份,硫辛酸30-45份,米糠粕880-920份。
3.权利要求1所述的添加剂在制备高植物蛋白源的鲟鱼配合饲料中的应用。
4.权利要求3所述的应用中的鲟鱼配合饲料,由下述组分按重量份组成:
组分 重量份
鱼粉 10-15
植物蛋白1 7-7.5
豆粕 18-26
菜粕 7.5-8.0
花生粕 3.5-8.0
玉米蛋白粉 11-21
面粉 9-12
沸石粉 2-2.5
维生素预混料 1
矿物质预混料 1
赖氨酸 0.05-0.075
磷酸二氢钙 1.5
鱼油 3
豆油 4-7
氯化胆碱 0.25
所述的植物蛋白1为大豆分离蛋白或膨化大豆。
5.权利要求4所述的配合饲料在制备成促进鲟鱼生长,提高鱼体蛋白合成和脂肪代谢,增强鲟鱼免疫力的饲料中的应用。
6.根据权利要求5所述的应用,所述的鲟鱼为杂交鲟。
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CN103636947A (zh) * | 2013-12-09 | 2014-03-19 | 浙江大学 | 用于高蛋白、低鱼粉水产配合饲料的饲料添加剂及其生产方法 |
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