CN114711344A - 一种促进鳗鲡生长及体表颜色变黄的方法 - Google Patents
一种促进鳗鲡生长及体表颜色变黄的方法 Download PDFInfo
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Abstract
本发明涉及一种促进鳗鲡生长及体表颜色变黄的方法,将小球藻粉用于鳗鲡饲料中部分替代鱼粉,制成小球藻配合饲料喂养鳗鲡,从而促进鳗鲡生长及体表颜色变黄。将小球藻配合饲料喂养鳗鲡30‑60天后,鳗鲡增重率可提高5‑25%、鱼体黄色值(b*)可达10‑30,鱼皮叶黄素含量可达1.0‑20.0μg/g。采用本发明方法养殖鳗鲡可减少鱼粉用量,同时还可增强鱼体免疫及代谢能力,以及提高鳗鲡生长速度和鱼体黄色调,从而有利于改善鳗鲡的营养价值及商品化价值,具有较大的应用前景。
Description
技术领域
本发明属于鱼类饲料技术领域,具体涉及一种促进鳗鲡生长及体表颜色变黄的方法。
背景技术
鳗鱼为高级食用鱼类,具有很高的食用价值,其肉质细嫩、味鲜美,深受广大消费者的喜爱。近年来世界鳗鱼养殖产量呈不断增长的趋势,总产量基本稳定在23万吨左右,中国鳗鱼养殖总产量居世界第一,约占世界总产量的70%。
全球养殖业的发展,使得鱼粉的需求量不断增加。然而,全球鱼粉资源有限、鱼粉价格不断上涨,研制和应用低鱼粉水产饲料成为水产饲料行业持久关注和追求的目标。因此,为了水产饲料的可持续生产,寻求一种可替代鱼粉并且具有较高营养价值、绿色可持续发展的原料对于水产饲料行业的变革具有重大的意义。
微藻是一种营养价值高且经济可行的鱼粉替代品,将其用于水产饲料可以降低对鱼粉的需求、减轻鱼粉生产的压力。微藻富含蛋白质、碳水化合物、脂肪酸、维生素、类胡萝卜素等营养物质,且各种氨基酸配比均衡,将其应用在水产饲料以部分替代鱼粉不仅可为水产动物提供充足的营养,还具有着色、增强鱼体免疫力等功效。
鳗鱼的体色在销售中的作用举足轻重,养出体色好的鳗鱼是所有养殖业者热切期望的,也是很多饲料制造厂一直在探索的课题,鳗鱼体色的改善能够极大地提高鳗鱼的经济价值。正常的鳗鱼背部为青黑色,腹部呈暗白色,而黄色调鳗鱼的商品价格远高于普通黑色鳗鱼,可极大提高鳗鱼的经济价值。然而截止目前,尚没有一种可减少饲料中鱼粉用量,同时又可提高鳗鲡生长及鱼体黄色调的方法。CN 102771680名称为“花鳗鲡幼鳗鱼阶段粉状配合饲料”的发明专利,该配合饲料配方由鱼粉、预糊化淀粉、发酵豆粕等组成。该专利的缺点是,鱼粉含量占比高,不能满足水产养殖产业可持续发展的需求。因此,有必要结合鳗鱼营养需求和体色调控的特点,通过精选原料,优化营养组成,开发出一种既能替代鱼粉又能够改善鳗鱼体色的配合饲料,以提高鳗鱼的经济价值,促进产业健康、可持续发展。
发明内容
针对以上不足,本发明的目的在于提供一种促进鳗鲡生长及体表颜色变黄的方法,在减少饲料中鱼粉用量的同时增强鱼体免疫及代谢能力,从而能够在较短喂养周期内显著提高鳗鲡生长及鱼体黄色调,实现改善鳗鲡的营养价值及商品化价值。
本发明通过以下技术方案达到上述目的:
一种促进鳗鲡生长及体表颜色变黄的方法,将小球藻粉用于鳗鲡配合饲料中部分替代鱼粉,制成小球藻配合饲料喂养鳗鲡,从而促进鳗鲡生长及体表颜色变黄。
进一步的,上述的小球藻粉中的蛋白质含量为48-58%、叶黄素含量为0.6-1.1%、类胡萝卜素含量为1.0-1.8%。
进一步的,上述的部分替代具体为:按照等量蛋白替代的原则,加入的小球藻粉的蛋白质的质量与替换掉的鱼粉的蛋白质的质量相等,被替代的鱼粉质量为鳗鲡配合饲料中鱼粉总质量的 5~20%。
进一步的,按占饲料总重的重量百分比含量计,上述的小球藻配合饲料中含有小球藻粉4.5-18%、进口白鱼粉52-62%、大豆浓缩蛋白2%、面粉1%、α-淀粉20-25%、啤酒酵母2%、蛋氨酸0.2%、复合多维0.2%、复合矿物质0.5%、氯化胆碱0.5%、卵磷脂1%、肝末粉1%、鱼油0.2%、豆油0-1.1%、乌贼膏1%。
进一步的,上述的小球藻配合饲料中,所述的复合多维由维生素A、维生素D3 、维生素E、维生素K3、维生素B1、维生素B2 、维生素B6 、维生素B12 、维生素C 、烟酰胺、泛酸钙、叶酸、生物素按质量比120:2:60:320:960:80:8:4000:320:160:40:1.6组成;所述的复合矿物质由硫酸亚铁、硫酸锰、碘酸钾、氯化钴、硫酸铜、硫酸锌、亚硒酸钠、硫酸镁按质量比34:3.161:0.278:0.441:3.768:12.347:0.055:37.143组成。
进一步的,上述的小球藻配合饲料的制备方法包括如下步骤:按占饲料总重的重量百分比含量计,将小球藻粉4.5-18%、进口白鱼粉52-62%、大豆浓缩蛋白2%、面粉1%、α-淀粉20-25%、啤酒酵母2%、蛋氨酸0.2%、复合多维0.2%、复合矿物质0.5%、氯化胆碱0.5%、卵磷脂1%、肝末粉1%、鱼油0.2%、豆油0-1.1%、乌贼膏1%的原料进行混合;混好的料先进行粗粉碎,然后经过超微粉碎;用80目的筛子进行筛选;筛选合格后再次混合,即得配合饲料。
采用上述的小球藻配合饲料喂养鳗鲡30-60天,鳗鲡增重率可提高5-25%。
采用上述的小球藻配合饲料喂养鳗鲡30-60天,鳗鲡鱼体黄色值(b*)可达10-30,鱼皮叶黄素含量可达1.0-20.0 μg/g。
与现有技术相比,本发明的有益效果在于:
1)采用富含蛋白质的小球藻粉部分替代鱼粉,不仅能减少鱼粉用量,还能满足鳗鲡对蛋白质等多种营养物质的需求,从而增强鱼体免疫及代谢能力,促进鳗鲡生长。
2)小球藻富含叶黄素等类胡萝卜素,采用其部分替代鳗鲡饲料中的鱼粉,可使鳗鲡摄入天然叶黄素实现在鱼皮中富集沉积,从而提高鱼体黄色调,可明显改善鳗鲡的营养价值及商品化价值。
附图说明
图1:本发明检测的鳗鲡背鳍前缘上方及侧线上方之间区域的色度值。
图2:本发明检测的鳗鲡背鳍后缘上方及侧线上方之间区域的色度值。
图3:本发明检测的鳗鲡臀鳍前缘上方及侧线下方之间区域的色度值。
图4:本发明检测的鳗鲡臀鳍后缘上方及侧线下方之间区域的色度值。
图5:本发明检测的鳗鲡皮肤叶黄素含量。
图6:本发明检测的鳗鲡血清及肝脏总蛋白含量。
图7:本发明检测的鳗鲡血清及肝脏白蛋白含量。
图8:本发明检测的鳗鲡血清及肝脏总胆固醇含量。
图9:本发明检测的鳗鲡血清及肝脏谷丙转氨酶含量。
图10:本发明检测的鳗鲡血清及肝脏谷草转氨酶含量。
图11:本发明检测的鳗鲡血清及肝脏总抗氧化能力。
图12:本发明检测的鳗鲡血清及肝脏过氧化氢酶含量。
图13:本发明检测的鳗鲡血清及肝脏碱性磷酸激酶含量。
图14:本发明检测的鳗鲡血清及肝脏总超氧化物歧化酶含量。
具体实施方式
下面结合附图对本发明的实施例作详细说明:本实施例在以本发明技术方案为前提下进行实验,给出了详细的实施方式和具体的操作过程,但本发明的保护范围不限于下述的实施例。
实施例1
小球藻粉的叶黄素、总类胡萝卜素和其他主要营养成分测定:
采用丙酮作为溶剂提取小球藻粉中的叶黄素、总类胡萝卜素,采用液相色谱法测定其含量;采用凯氏定氮法测定小球藻粉中的蛋白质含量。
表1 小球藻粉的叶黄素、总类胡萝卜素和其他主要营养成分
下述实施例中所采用小球藻粉均与本实施例相同。
实施例2
小球藻配合饲料配方和小球藻粉添加量的确定:
根据鳗鲡生长发育过程中的营养需求、原料特性和配方经验,确定饲料配方中的主要原料组成和小球藻粉加入量。以进口白鱼粉、大豆浓缩蛋白、面粉、α-淀粉、啤酒酵母、蛋氨酸、复合多维、复合矿物质、氯化胆碱、卵磷脂、肝末粉、鱿鱼油、豆油、乌贼膏配制鳗鲡配合饲料;其中,所述的进口白鱼粉是由鳕鱼、比目鱼等整鱼或下脚料经过蒸煮、提脂、烘干、粉碎等制成,粗蛋白含量为65%;所述的复合多维由维生素A、维生素D3 、维生素E、维生素K3、维生素B1、维生素B2 、维生素B6 、维生素B12 、维生素C 、烟酰胺、泛酸钙、叶酸、生物素按质量比120:2:60:320:960:80:8:4000:320:160:40:1.6组成;所述的复合矿物质由硫酸亚铁、硫酸锰、碘酸钾、氯化钴、硫酸铜、硫酸锌、亚硒酸钠、硫酸镁按质量比34:3.161:0.278:0.441:3.768:12.347:0.055:37.143组成。为了科学评估小球藻粉对鳗鲡生长发育及鱼体色调的影响,本实施例中设置1个对照组(未使用小球藻粉替代进口白鱼粉制得的鳗鲡饲料)和4个实验组(将小球藻粉用于鳗鲡饲料中部分替代进口白鱼粉制得的小球藻配合饲料),因此共5组饲料。其中,部分替代的方式为:按照等量蛋白替代的原则,加入的小球藻粉的蛋白质的质量与替换掉的鱼粉的蛋白质的质量相等。
具体饲料配方和测定营养成分见下表。其中,饲料1~5中被替代的鱼粉质量依次为鳗鲡配合饲料中鱼粉总质量的0%、 5%、10%、15%和20%。
表2 具体饲料配方和测定营养成分
实施例3
饲料加工:
按实施例2表2中配方称取饲料原料进行混合;混好的料先进行粗粉碎,然后经过超微粉碎;用80目的筛子进行筛选;筛选合格后再次混合,即得成品饲料。
实施例4
鳗鲡养殖试验:
选用同一批规格一致、机体健康的黑仔鳗苗作为实验鱼,平均体重为6.3±0.39g/尾。设置五组试验,分别为投喂饲料1、饲料2、饲料3、饲料4、饲料5,其中饲料1为空白对照。试验在15个相同规格(60cm*50cm*60cm)的长方形玻璃缸中,每组实验设置三个重复,每个玻璃缸放养15尾鱼,采用循环水养殖模式,养殖实验共进行60天。
养殖实验结束后,鳗鲡饥饿24 h后采样,统计其摄食及生长数据。
养殖实验结束后,每组随机选取5尾鳗鱼,并进行低温匀浆处理,采用GB 5009.5-2016 凯氏定氮法测定鳗鱼的粗蛋白含量,采用GB/T 5009.6-2003 索式抽提法测定鳗鱼的粗脂肪含量,采用GB 5009.3-2016 测定鳗鱼的水分含量,采用GB/T 5009.4-2003 测定鳗鱼的灰分含量,并测定鳗鱼体表色值和皮肤中叶黄素含量,以空白对照组的鳗鱼作为参照。
养殖实验结束后,每组随机取10尾鱼采用断尾取血于2mL离心管中,4 ℃静置6 h,低温离心(4 ℃,3000 r/min,10 min),取上清即得血清,并且在冰浴条件下快速解剖鳗鲡,取肝脏及鱼皮。准确称取肝脏重量,与生理盐水按1:9的比例混匀,采用组织匀浆机冰浴匀浆,匀浆液在4 ℃条件下3000 r/min离心10 min,取匀浆上清液装于5mL离心管中,样品采集完毕,及时保存于-80 ℃冰箱中待测。采用南京建成生物工程研究所试剂盒测定血清和肝脏的总胆固醇(Total cholesterol, T-CHO)、白蛋白(Albumin, ALB)、总蛋白(Totalprotein,TP)、谷草转氨酶(Glutamic oxalacetic transaminase,GOT)、谷丙转氨酶(Glutamate pyruvate transaminase,GPT)、总超氧化物歧化酶(Total superoxidedismutase,TSOD)、过氧化氢酶(Catalase,CAT)、总抗氧化能力(Total antioxidantcapacity,TAOC)碱性磷酸酶(Alkaline phosphatase,AKP)活性。
数据处理:采用IBM SPSS Statistics 26.0软件对数据进行单因素方差分析(One-way ANOVA),用Duncan多重比较分析组间差异显著性,用P<0.05时认为有显著性差异。
皮肤体色值测定:采用国际发光照明委员会规定的L、a*、b*色度空间表示鳗鱼体色,其中L表示亮度,+a表示红色,-a*表示绿色,+b*表示黄色,-b*表示蓝色。养殖实验结束后,每个玻璃缸随机测定3尾鱼。使用柯尼卡-美能达CR-400色差计测定体色,分别测定鳗鱼背部和腹部皮肤的体色值,背部分别为背鳍前缘上方及侧线上方之间的区域和背鳍后缘上方及侧线上方之间的区域,腹部分别为臀鳍前缘上方及侧线下方之间的区域和臀鳍后缘上方及侧线下方之间的区域。
皮肤叶黄素含量的测定:养殖实验结束后每个玻璃缸随机选取5尾鱼的鱼皮于-80℃冰箱中保存,用于测定皮肤中的叶黄素含量。取100 mg冻干后的鱼皮置于2 mL的离心管中,加入1 mL的60% w/w 的KOH溶液和 1 g研磨珠。用珠磨破碎仪破碎7 min,然后40℃水浴40 min。在通风厨中将水浴好的混合物倒入15 mL离心管中,用1 mL无水乙醚清洗破碎用的离心管中的色素,重复三次,合并清洗液加入混合物中,用震荡器震荡30秒后于4000 rpm离心2 min,将上层乙醚倒入棕色瓶中。加入3 mL乙醚进行萃取,重复以萃取步骤直至乙醚层无色。用氮气吹干萃取的乙醚液,加入1 mL丙酮,震荡30秒进行重悬。采用高效液相色谱法测定叶黄素的含量。具体所用仪器型号及色谱条件如下所示:
仪器:Agilent 1200 Series高效液相色谱仪;
色谱柱:YMC30 RP-30(4.6 mm×250 mm×5μm);
进样量:20μL;
检测波长:450 nm;
柱温:30℃;
流动相A:称取3.854 g乙酸铵和1 g丁基羟基甲苯溶解于800 mL甲醇,加入0.5 mL三乙胺和30 mL水,混匀,再用无水甲醇定容至1 L容量瓶,过滤,超声;
流动相B:称取0.5 g丁基羟基甲苯溶解于400 mL甲基叔丁基醚,加0.25 mL三乙胺,混匀,再加一定量甲基叔丁基醚定容至500 mL容量瓶,过滤,超声;
流动相洗脱条件:流速1 mL/min,按下表方法进行设定:
表3 洗脱条件
存活率、增重率、特定生长率、饲料系数及肥满度的计算方法为:
(1)存活率(Survival rate,SR,%)=100×N2/N1;
(2)增重率(Weight gain rate,WGR,%)=100×(Wt-W0)/W0;
(3)特定生长率(Specific grow rate,SGR,%/d)=100×(ln Wt-ln W0)/t;
(4)饲料系数(Feed conversion ratio,FCR)=F/(Wt-W0);
(5)肥满度(Condition factor CF,g/cm3)=100×Wb/L3。
式中:N2为每组实验结束时鱼尾数;N1为每组实验开始时鱼尾数;t为饲喂天数(d);W0为平均每组鱼初始总重(g/组);Wt为平均每组鱼终末总重(g/组);F为 每尾鱼摄食的饲料干物质重(g),Wb为鱼末重(g/尾);L为鱼体长(cm/尾)。
如表4所示,与空白对照组相比,饲料2、饲料3、饲料4、饲料5对鳗鲡的存活率无显著影响;而且添加了小球藻粉的试验组的增重率、特定生长率比对照组更高,饲料系数相对于对照组降低,表明投喂小球藻基饲料能提高鳗鱼的生长速度和饲料转换率,可以有效地提高产量、降低成本,具有很高的经济价值。
表4 小球藻基饲料对鳗鱼生长性能的影响
注:同行数值肩标英文大写字母不同表示P<0.01,同行数值肩标英文小写字母不同表示P<0.05,肩标字母相同或无标记为P>0.05。
如表5所示,与空白对照组相比,鳗鱼的粗蛋白含量随着小球藻用量的增加呈先增后减的趋势,鳗鱼粗脂肪含量的趋势则相反。饲料3试验组的粗蛋白含量显著高于空白对照组(P<0.05),各组水分含量均没有显著性差异,各组的粗灰分含量与对照组相比也无显著性差异。上述结果说明使用小球藻替代饲料中的鱼粉能增加鳗鱼的蛋白质含量,其中饲料2试验组的效果最好,极大地提高了鳗鱼的营养价值。
表5鳗鱼营养成分分析
注:同行数值肩标英文大写字母不同表示P<0.01,同行数值肩标英文小写字母不同表示P<0.05,肩标字母相同或无标记为P>0.05。
如图1~4所示,与空白对照组相比,饲料2~5试验组鳗鱼背部(分别为背鳍前缘上方及侧线上方之间的区域和背鳍后缘上方及侧线上方之间的区域)及腹部(分别为臀鳍前缘上方及侧线下方之间的区域和臀鳍后缘上方及侧线下方之间的区域)的黄色值(b*)均显著提高,说明本发明提供的实施例饲料能够显著提高鳗鱼鱼体的黄色调。
如图5所示,与对照组相比,饲料2~5试验组的鳗鱼皮叶黄素含量均显著提升,并且鳗鱼皮的叶黄素含量随着鳗鱼饲料中小球藻替代鱼粉比例的增加而增加,即饲料5试验组的鳗鱼皮的叶黄素含量最高。
如图6~7所示,血清总蛋白、血清白蛋白及肝脏白蛋白的含量均随小球藻添加量的增加呈先增后减的趋势,饲料2~4试验组鳗鲡的血清总蛋白含量显著高于对照组(P<0.05),饲料3、饲料5试验组的血清白蛋白含量显著高于对照组(P<0.05),饲料3~5试验组鳗鲡的肝脏白蛋白含量显著高于对照组(P<0.05)。血液中的总蛋白由肝脏合成,血清总蛋白含量升高代表鳗鱼肝脏合成蛋白质能力增强,白蛋白参与维持水产动物血清中的渗透压并能以胶体的形式对蛋白起到保护作用,上述结果表明小球藻基饲料组的机体抗病力高于对照组。
如图8所示,饲料2~5试验组鳗鲡的血清总胆固醇含量和对照组相比无显著差异,然而肝脏的总胆固醇含量均显著高于对照组(P<0.05),结合小球藻基饲料组优于对照组的生长性能,表明在一定的范围内肝脏胆固醇含量的增加可能与鳗鱼生长性能的提升有关。总胆固醇是血清脂类的重要组成,反应机体脂类代谢的程度,上述结果表明小球藻基饲料组与对照组的脂类代谢水平基本一致。
如图9~10所示,血清谷丙转氨酶含量随小球藻添加量的增加呈先上升后下降的趋势;饲料2~5试验组鳗鲡的肝脏的谷丙转氨酶含量均显著高于对照组(P<0.05),但血清的谷草转氨酶含量均显著低于对照组(P<0.05),肝脏的谷草转氨酶含量均显著高于对照组(P<0.05)。谷丙转氨酶和谷草转氨酶是水产动物机体中参与氨基酸代谢的重要酶类,在健康情况下主要存在肝脏中,血清中含量较低。上述结果表明,小球藻基饲料组肝脏的氨基酸代谢能力比对照组强。
如图11~14所示,饲料3、饲料5试验组鳗鲡的血清总抗氧化能力均显著高于对照组(P<0.05),且饲料2~5试验组鳗鲡的肝脏总抗氧化能力均显著高于对照组(P<0.05);饲料2~5试验组鳗鲡的血清过氧化氢酶含量均显著高于对照组(P<0.05),且实施例饲料2~3试验组鳗鲡的肝脏过氧化氢酶含量显著高于对照组(P<0.05);饲料2~5试验组鳗鲡的血清碱性磷酸酶含量与对照组无显著性差异,而饲料2~4试验组鳗鲡的肝脏碱性磷酸酶含量显著高于对照组;饲料2~5试验组鳗鲡的血清超氧化物歧化酶的含量均高于对照组,而肝脏碱性磷酸酶含量与对照组无显著性差异。根据上述结果表明,小球藻基饲料组的免疫力及抗氧化能力均高于对照组。
综上所述,在试验范围内使用小球藻粉部分替代鳗鲡饲料中鱼粉均可达到提高鳗鲡生长和鱼体黄色调的效果,且随替代比例的提高其效果越明显。
以上所述仅为本发明的较佳具体实施例,但本发明的保护范围并不局限与此,凡依本发明申请专利范围所做的均等变化及修饰,且任何熟悉本技术领域的技术人员在本发明揭露的技术范围内,根据本发明的技术方案及其构思加以等同替换或改变,皆应属本发明的涵盖范围。
Claims (6)
1.一种促进鳗鲡生长及体表颜色变黄的方法,其特征在于:将小球藻粉用于鳗鲡饲料中部分替代鱼粉,制成小球藻配合饲料喂养鳗鲡,从而促进鳗鲡生长及体表颜色变黄。
2.根据权利要求1所述的一种促进鳗鲡生长及体表颜色变黄的方法,其特征在于:所述的小球藻粉中的蛋白质含量为48-58%、叶黄素含量为0.6-1.1%、类胡萝卜素含量为1.0-1.8%。
3.根据权利要求1所述的一种促进鳗鲡生长及体表颜色变黄的方法,其特征在于:按照等量蛋白替代的原则,加入的小球藻粉的蛋白质的质量与替换掉的鱼粉的蛋白质的质量相等,被替代的鱼粉质量为鳗鲡饲料中鱼粉总质量的 5~20%。
4.根据权利要求3所述的一种促进鳗鲡生长及体表颜色变黄的方法,其特征在于:按占饲料总重的重量百分比含量计,所述的小球藻配合饲料中含有小球藻粉4.5-18%、进口白鱼粉52-62%、大豆浓缩蛋白2%、面粉1%、α-淀粉20-25%、啤酒酵母2%、蛋氨酸0.2%、复合多维0.2%、复合矿物质0.5%、氯化胆碱0.5%、卵磷脂1%、肝末粉1%、鱼油0.2%、豆油0-1.1%、乌贼膏1%。
5.根据权利要求4所述的一种促进鳗鲡生长及体表颜色变黄的方法,其特征在于:所述的复合多维由维生素A、维生素D3 、维生素E、维生素K3、维生素B1、维生素B2 、维生素B6 、维生素B12 、维生素C 、烟酰胺、泛酸钙、叶酸、生物素按质量比120:2:60:320:960:80:8:4000:320:160:40:1.6组成;所述的复合矿物质由硫酸亚铁、硫酸锰、碘酸钾、氯化钴、硫酸铜、硫酸锌、亚硒酸钠、硫酸镁按质量比34:3.161:0.278:0.441:3.768:12.347:0.055:37.143组成。
6.根据权利要求4所述的一种促进鳗鲡生长及体表颜色变黄的方法,其特征在于:所述的小球藻配合饲料的制备方法包括如下步骤:按占饲料总重的重量百分比含量计,将小球藻粉4.5-18%、进口白鱼粉52-62%、大豆浓缩蛋白2%、面粉1%、α-淀粉20-25%、啤酒酵母2%、蛋氨酸0.2%、复合多维0.2%、复合矿物质0.5%、氯化胆碱0.5%、卵磷脂1%、肝末粉1%、鱼油0.2%、豆油0-1.1%、乌贼膏1%的原料进行混合;混好的料先进行粗粉碎,然后经过超微粉碎;用80目的筛子进行筛选;筛选合格后再次混合,即得小球藻配合饲料。
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