CN116602251A - 一种用于提高南美白对虾的虾青素酯含量的养殖方法 - Google Patents
一种用于提高南美白对虾的虾青素酯含量的养殖方法 Download PDFInfo
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- CN116602251A CN116602251A CN202310705659.6A CN202310705659A CN116602251A CN 116602251 A CN116602251 A CN 116602251A CN 202310705659 A CN202310705659 A CN 202310705659A CN 116602251 A CN116602251 A CN 116602251A
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- China
- Prior art keywords
- penaeus vannamei
- feed
- astaxanthin
- powder
- vannamei boone
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/50—Culture of aquatic animals of shellfish
- A01K61/59—Culture of aquatic animals of shellfish of crustaceans, e.g. lobsters or shrimps
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/179—Colouring agents, e.g. pigmenting or dyeing agents
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
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Abstract
本发明提供一种用于提高南美白对虾的虾青素酯含量的养殖方法,使用指定的饲料投喂,能够有效提高对虾体内的虾青素酯含量。本发明还提供了一种具有提高南美白对虾体内虾青素酯含量的饲料,其为添加有3%坛紫菜粉的南美白对虾基础饲料和包含有添加有0.1%玉米黄素或0.1%岩藻黄素的饲料。使用上述两种饲料对南美白对虾进行投喂,能效提高对虾体内的虾青素酯含量。
Description
技术领域
本发明属于水产养殖技术领域,具体涉及一种用于提高南美白对虾的虾青素酯含量的养殖方法。
背景技术
南美白对虾(学名:Litopenaeus Vannamei)是对虾科、对虾属动物,具有生长快、肉质鲜美、营养价值高等特点,属于世界上养殖产量最高的虾类之一。自引入我国后,得到大规模的推广养殖,约占我国对虾养殖总产量的80%以上,成为我国养殖产量最高的对虾养殖品种。
虾青素主要以虾青素酯的形式存在,广泛存在于水产生物中,具有组织着色、提高环境适应能力、参与自由基清除、增强繁殖功能及提高机体免疫力等功能。虾青素酯在对虾形成颜色时特别重要,市面上南美白对虾的销售价格,99%是由虾身颜色决定的。虾青素也是重要的抗氧化剂,强大的抗氧化能力有助于保护虾的营养储备,增强繁殖功能,降低血清中的脂质。同时,虾青素酯也可改善血淋巴免疫指标,提高免疫力和抗应激能力。
在自然环境条件下,虾不能自身合成虾青素,只能通过食用浮游动物、甲壳或海藻,利用酮化和羟基化步骤将膳食类胡萝卜素进行转化或直接吸收成虾青素及其酯化形式。目前的南美白对虾养殖过程中,多通过在池塘内直接补充藻类作为虾青素来源,而藻类的恣意生长会产生水体酸碱度的变化、有毒气体等影响对虾的健康,给养殖户的管理增加难度。此外,饲料中直接补充虾青素也是提高水产动物中虾青素酯含量的直接办法,但由于虾青素原料昂贵,导致养殖成本极增。海藻是理想的饲料组成,具有产值高、富含类胡萝卜素以及附加多种活性物质,是虾青素酯形成的重要来源,既能满足南美白对虾营养需求,又增强其品质和抗病性。因此,发展藻源类胡萝卜素饲料,促进对虾养殖业健康发展,为消费者提供营养丰富的高品质对虾。
发明内容
本发明的目的是提供一种用于提高南美白对虾的虾青素酯含量的养殖方法,用于提高南美白对虾体内的虾青素酯含量。
本发明首先提供了一种具有提高南美白对虾体内虾青素酯含量的饲料,所述的饲料为添加有3%坛紫菜粉的南美白对虾基础饲料;
所述的坛紫菜粉,是将坛紫菜用淡水清洗,除去杂藻、晒干除水、剪碎和高温烘烤,研磨成粉制备的。
本发明还提供了一种提高南美白对虾的虾青素酯的养殖方法,是使用上述的饲料进行投喂;
作为实施例的一种具体记载,投喂量为对虾体重的5%—8%;
更进一步的,所投喂的饲料还包含有添加有0.1%玉米黄素或0.1%岩藻黄素的饲料。
本发明再筛选获得了能够有效提高南美白对虾的虾青素酯含量的饲料的基础上,提供一种南美白对虾的养殖方法,能够有效提高对虾体内的虾青素酯含量。
附图说明
图1:南美白对虾中虾青素酯的选择离子流图,其中A为Am-C16:0;B为Am-C18:0;C为Ad-C20:5/16:0;D为Ad-C22:6/16:0;E为Ad-C 20:5/20:5;F为Ad-C 22:6/18:1;G为Ad-C22:6/20:5;H为Ad-C 22:6/22:6。
图2:虾青素酯的质谱裂解途径图。
图3:不同藻粉添加组中南美白对虾的虾壳、肝胰腺和肌肉的虾青素酯组成图
图4:0.1%玉米黄素和0.1%岩藻黄素组中南美白对虾的虾壳、肝胰腺和肌肉中的虾青素酯组成图。
图5:不同坛紫菜添加比例组中南美白对虾的虾壳、肝胰腺和肌肉的虾青素酯组成图。
具体实施方式
本发明所使用南美白对虾幼虾由浙江省宁波大学周岐存教授提供。选择海带粉、羊栖菜粉、坛紫菜、玉米黄素和岩藻黄素作为饲料添加剂,按照不同添加比例共配制10种南美白对虾饲料。在基础饲料的基础上,分别添加3%海带粉(TL-3%)、3%羊栖菜粉(SF-3%)、1%坛紫菜粉(PH-1%)、2%坛紫菜粉(PH-2%)、3%坛紫菜粉(PH-3%)、4%坛紫菜粉(PH-4%)、5%坛紫菜粉(PH-5%)、0.1%玉米黄素(ZX-01%)和0.1%岩藻黄素(FX-0.1%)替代纤维素,配制成相应的饲料,分为不同藻粉组、不同比例坛紫菜粉组和不同类胡萝卜素组。不添加藻粉或类胡萝卜素的基础饲料为对照组。幼虾经基础饲料暂养两周后,挑选体色、规格一致的健康南美白对虾幼虾(均重2.94±0.14g)。将幼虾随机分成10组,每组3重复,每重复30尾虾。实验设置空白组、不同藻粉组、不同比例坛紫菜粉组和不同类胡萝卜素组。将30个养殖桶中的南美白对虾用基础饲料驯化一周,然后分别用上述10组饲料进行投喂,养殖时间为8周。每天投喂三次(6:30、12:30、18:30),日投饵量约占对虾体重的5%—8%,早晚投喂量约占总投喂量的70%,并根据实际情况做适当调整。每隔2d吸污与换水,保持水质。养殖期间水温控制为22~35℃、氨氮应在0.2mg/L以下,亚硝酸盐在0.1mg/L以下,硫化氢在0.1mg/L以下,pH值范围为7.8~8.8,溶氧在4.0mg/L以上。
8周养殖实验结束后,分别取南美白对虾的虾壳、肌肉、肝胰腺作为实验样本。每个平行取4mL离心管的肝胰腺、7只虾壳和7只虾肌肉,各3个平行,然后迅速迅速放入液氮中冻干,然后转移至-80℃冰箱保存。
下面结合具体实施例和附图对本发明进行详细的描述。
实施例1:藻源饲料加工制备
选择海带粉、羊栖菜粉、坛紫菜、玉米黄素和岩藻黄素作为饲料添加剂,按照不同添加比例共配制10种南美白对虾饲料。在基础饲料的基础上,分别添加3%海带粉(TL-3%)、3%羊栖菜粉(SF-3%)、1%坛紫菜粉(PH-1%)、2%坛紫菜粉(PH-2%)、3%坛紫菜粉(PH-3%)、4%坛紫菜粉(PH-4%)、5%坛紫菜粉(PH-5%)、0.1%玉米黄素(ZX-01%)和0.1%岩藻黄素(FX-0.1%)替代纤维素,配制成相应的饲料,分为三组饲料:不同藻粉组、不同比例坛紫菜粉组和不同类胡萝卜素组。不添加藻粉或类胡萝卜素的基础饲料为对照组。所有饲料原料经小型粉碎机粉碎后过80目筛,过筛饲料原料按比例称重后运用逐级放大法将饲料原料均匀混合,并使用饲料制粒机将饲料制成直径为1mm和1.5mm的硬颗粒饲料,在60℃烘箱中将饲料烘干至恒重。饲料配方及其营养水平见表1和表2。
表1:不同南美白对虾饲料配方及营养组成表
注:每kg预混料提供矿物质元素:铁6g;铜0.3g;锰1.5g;锌4g;硒0.02g;碘0.07g;钴0.007g;提供维生素:维生素A 375000IU;维生素D3 150000IU;维生素E 4000mg;维生素K3 400mg;维生素B1 600mg;维生素B2 1000mg;维生素B6 850mg;维生素B12 2mg;烟酸10000mg;叶酸200mg;肌醇12000mg;D-生物素8mg;D-泛酸钙2200mg;维生素C 14000mg。
表2:添加不同种类藻粉/类胡萝卜素的饲料配方及营养组成表
注:每kg预混料提供矿物质元素:铁6g;铜0.3g;锰1.5g;锌4g;硒0.02g;碘0.07g;钴0.007g;提供维生素:维生素A 375000IU;维生素D3 150000IU;维生素E 4000mg;维生素K3 400mg;维生素B1 600mg;维生素B2 1000mg;维生素B6 850mg;维生素B12 2mg;烟酸10000mg;叶酸200mg;肌醇12000mg;D-生物素8mg;D-泛酸钙2200mg;维生素C 14000mg。
实施例2:确定具有增强南美白对虾虾青素含量的藻粉种类
将上述不同藻粉组南美白对虾中的虾壳、肌肉、肝胰腺组织进行虾青素酯提取。
将不同组织样品冷冻干燥,并在液氮下研磨成细粉末,准确称取500mg肝胰腺、肌肉或虾壳样品,放置于EP管中,并加入5mL甲醇溶液(含0.1%BHT)作为提取液。将EP管震荡30s后,转移至超声仪中,超声提取30min;再加入5mL二氯甲烷溶液(含0.1%BHT),继续震荡30s,超声提取30min。将提取液放置于冰上静置5min,在12000r/min条件下高速离心30min,取上清液并放置于新的EP管中,并用旋转浓缩仪将上清液浓缩近干。将其转移至进样瓶中,并用甲醇溶液(含0.1%BHT)定容至1.0mL,使用0.22μm滤膜过滤,得到滤液,待分析。整个实验处理过程皆在低温、避光的环境下进行。
在南美白对虾肌肉、虾壳和肝胰腺中共检测到8个虾青素酯,其母离子和特征碎片离子见表3,选择离子流图和质谱裂解规律如图1和图2所示。采用亚油酸虾青素单酯(Am-C18:2)和双棕榈酸虾青素双酯(Ad-C16:0/16:0)标准品进行虾青素酯含量分析。以纵坐标为标准品峰面积,横坐标为浓度,绘制标准工作曲线,得到亚油酸虾青素单酯(Am 18:2)和双棕榈酸虾青素双酯(Ad-C16:0/16:0)的线性方程式分别为Y=684594*X+63328和Y=796664*X+46077,依据该标准曲线对南美白对虾不同组织中的虾青素酯进行定量分析。
表3:南美白对虾中虾青素酯的母离子和特征碎片离子信息表
如图3所示,在TL-3%组、SF-3%组和PH-3%组的南美白对虾虾壳、肝胰腺和肌肉中共检测到了6、4和2种虾青素酯,其中虾壳和肝胰腺中的虾青素酯主要以双酯形式存在,均检测到Ad-C22:6/16:0、Ad-C22:6/18:1、Ad-C22:6/20:5和Ad-C22:6/22:6,而肌肉中主要以单酯形式存在。此外,虾青素酯的总含量在虾壳、肝胰腺和肌肉分别为217.50mg/kg、41.18mg/kg、116.82mg/kg,虾壳中的含量分别是肝胰腺和肌肉的5.2和1.8倍。由此表明,虾壳中虾青素酯数量和含量最多,是主要的虾青素酯积累部位,而肌肉以累积虾青素单酯为主。
在虾壳中,6种虾青素酯(Ad-C20:5/16:0、Ad-C22:6/16:0、Ad-C20:5/20:5、Ad-C22:6/18:1、Ad-C22:6/20:5、Ad-C22:6/22:6)的含量随着藻粉种类的变化而呈现出不同的变化趋势,整体而言,TL-3%组、SF-3%和PH-3%组的总虾青素酯含量是对照组的1.5、0.9和3.5倍,即PH-3%组>TL-3%组>对照组>SF-3%。此外,PH-3%组中的6种虾青素酯的含量分别是对照组的3.4、3.6、4.1、3.7、3.5和3.9倍,根据其含量由高到低顺序为:Ad-C22:6/16:0、Ad-C22:6/20:5、Ad-C22:6/22:6、Ad-C22:6/18:1、Ad-C20:5/16:0、Ad-C20:5/20:5。在肝胰腺中,只有PH-3%组的虾青素酯总含量高于对照组,是对照组的4.9倍,Ad-C22:6/18:1和Ad-C22:6/22:6的含量分别占了四种虾青素酯的31.9%和37.6%,为其主要的虾青素酯;然而,TL-3%和SF-3%的总虾青素酯含量则显著低于对照组,仅是对照组的70.1%、20.2%。在肌肉中,仅检测到了2种虾青素单酯,分别为Am-C16:0和Am-C18:0,其中Am-C16:0是主要的虾青素单酯,占了总虾青素酯的67.4%。两种虾青素单酯在三种藻粉添加剂组中的含量顺序为:PH-3%组>TL-3%组≈SF-3%>对照组,分别是对照组的2.2倍、1.4倍和1.3倍。结果表明3%坛紫菜粉作为饲料添加剂对南美白对虾中的虾青素酯含量提升效果最优。
实施例3:不同类胡萝卜素饲料添加剂对南美白对虾虾青素含量的影响
用实施例2中的方法对不同类胡萝卜素组虾青素含量进行检测,如图4所示,将含0.1%玉米黄素和0.1%岩藻黄素作为饲料添加剂进行南美白对虾投喂实验,分别在肌肉、虾壳和肝胰腺中检测到了3、6和2种虾青素酯,其中虾壳中的虾青素酯数量最多,其次是肌肉,肝胰腺中的虾青素酯含量最少。此外,虾壳、肌肉和肝胰腺中虾青素组成也存在较大差异,其中虾壳和肝胰腺中的虾青素酯主要以双酯形式存在,在虾壳和肝胰腺均检测到Ad-C22:6/16:0、Ad-C22:6/18:1、Ad-C22:6/20:5和Ad-C22:6/22:6,而肌肉中主要以单酯形式存在。虾青素酯的总含量,在虾壳、肝胰腺和肌肉分别为91.9646mg/kg、29.1087mg/kg、53.1536mg/kg,虾壳中的含量分别是肝胰腺和肌肉的3.1和1.7倍。
在虾壳中,根据不同类胡萝卜素的添加,6种虾青素酯(Ad-C20:5/16:0、Ad-C22:6/16:0、Ad-C20:5/20:5、Ad-C22:6/18:1、Ad-C22:6/20:5、Ad-C22:6/22:6)的含量呈现不同的高低趋势,其中ZX-0.1%组的虾青素酯的总含量最高,ZX-0.1%组和FX-0.1%组的虾青素酯总含量分别为对照组的1.2和1.0倍。在肝胰腺中,根据不同藻粉的添加,4种虾青素酯的含量呈现不同的高低趋势,其中ZX-0.1%组的虾青素酯的总含量最高,ZX-0.1%组和FX-0.1%组的虾青素酯总含量分别为对照组的2.3和1.9倍。在肌肉中检测到的2种虾青素单酯,即Am-C16:0和Am-C18:0,含量差距较大,其中Am-C16:0为主要的虾青素单酯,占了总虾青素酯的91.0%。FX-0.1%组的虾青素酯的总含量最高,ZX-0.1%组和FX-0.1%组的虾青素酯总含量分别为对照组的1.1和1.2倍。结果证实,0.1%玉米黄素和0.1%岩藻黄素作为饲料添加剂提高了南美白对虾虾青素的含量,但效果较3%坛紫菜粉差。
实施例4:确定具有增强南美白对虾虾青素含量的藻粉比例
用实施案例1中所制备的不同比例坛紫菜粉饲料分别投喂南美白对虾,每天投喂三次(6:30、12:30、18:30),日投饵量约占对虾体重的5%—8%,早晚投喂量约占总投喂量的70%。每隔2d吸污与换水,保持水质。
用实施例2中的方法对不同比例坛紫菜粉组虾青素含量进行检测。如图5所示,在PH-1%~PH-5%组的南美白对虾虾壳、肝胰腺和肌肉中检测到了6、4和2种虾青素酯,同样虾壳和肝胰腺中的虾青素酯主要以双酯形式存在,Ad-C22:6/16:0、Ad-C22:6/18:1、Ad-C22:6/20:5和Ad-C22:6/22:6在虾壳和肝胰腺均检测到,而肌肉中主要以单酯形式存在。虾壳、肝胰腺和肌肉中的虾青素酯总含量分别为303.0063mg/kg、106.8159mg/kg、70.3004mg/kg,虾壳中的含量分别是肝胰腺和肌肉的2.8和4.3倍。
在虾壳中,随着坛紫菜粉比例的增加,6种虾青素酯(Ad-C20:5/16:0、Ad-C22:6/16:0、Ad-C20:5/20:5、Ad-C22:6/18:1、Ad-C22:6/20:5、Ad-C22:6/22:6)的含量呈现先增加后减少的趋势,分别为对照组的3.4、3.5、4.1、3.7、3.5和3.9倍,且在PH-3%组的含量达到了最高值,其虾青素酯总含量为对照组的3.7倍。在肝胰腺中检测到的4种虾青素酯(Ad-C22:6/16:0、Ad-C22:6/18:1、Ad-C22:6/20:5、Ad-C22:6/22:6)的含量随着坛紫菜粉比例的增加呈现先增加后减少的趋势,且在PH-4%组的含量达到了最高值,其中Ad-C22:6/18:1和Ad-C22:6/22:6为主要的虾青素酯,分别占了四种虾青素酯的30.9%和36.8%。在肌肉中同样检测到了2种虾青素单酯,即Am-C16:0和Am-C18:0,2种虾青素单酯的含量同样也呈现先增加后减少的趋势,且在PH-3%组的含量达到了最高值,即两种虾青素单酯在五个不同比例坛紫菜粉添加剂组中的含量顺序为:PH-3%组>PH-4%组≈PH-2%≈PH-5%>PH-1%组>对照组,分别为对照组的2.7和1.4倍。
结果证实,不同比例坛紫菜粉添加剂均可以提高南美白对虾虾壳、肝胰腺和肌肉中虾青素酯的含量,但随着紫菜粉比例的增加,虾青素酯的含量到达一个峰值后开始呈现下降趋势,3%坛紫菜粉提升效果最优。
Claims (7)
1.一种用于提高南美白对虾的虾青素酯含量的养殖方法,其特征在于,所述的方法,是使用添加3%坛紫菜粉的南美白对虾基础饲料来投喂南美白对虾。
2.如权利要求1所述的方法,其特征在于,所述的方法,还投喂添加有0.1%玉米黄素或0.1%岩藻黄素的南美白对虾基础饲料。
3.如权利要求1所述的方法,其特征在于,所述的坛紫菜粉,是将坛紫菜用淡水清洗,除去杂藻、晒干除水、剪碎和高温烘烤,研磨成粉制备的。
4.如权利要求1所述的方法,其特征在于,所述的方法,投喂量为对虾体重的5%-8%。
5.一种用于提高南美白对虾的虾青素酯含量的饲料,其特征在于,所述的饲料是添加3%坛紫菜粉的南美白对虾基础饲料。
6.如权利要求5所述的用于提高南美白对虾的虾青素酯含量的饲料,其特征在于,所述的饲料中还添加有0.1%玉米黄素。
7.如权利要求5或5所述的用于提高南美白对虾的虾青素酯含量的饲料,其特征在于,所述的饲料中还添加有0.1%岩藻黄素。
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