CN113812545A - 一种调控高脂投喂下大黄鱼肌肉脂肪酸组成的营养学方法 - Google Patents
一种调控高脂投喂下大黄鱼肌肉脂肪酸组成的营养学方法 Download PDFInfo
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- CN113812545A CN113812545A CN202111100758.9A CN202111100758A CN113812545A CN 113812545 A CN113812545 A CN 113812545A CN 202111100758 A CN202111100758 A CN 202111100758A CN 113812545 A CN113812545 A CN 113812545A
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Abstract
本发明涉及一种调控高脂投喂下大黄鱼肌肉脂肪酸组成的营养学方法，属于水产营养饲料领域，在大黄鱼高脂饲料中添加含量为0.5％的植物甾醇。本发明方法能够显著降低肌肉脂肪酸组成中SFA的含量比例，并在一定程度上提高n‑6PUFA和n‑3PUFA的含量比例，提高鱼肉的营养价值。
Description
技术领域
本发明属于水产动物营养饲料领域，具体地涉及一种调控大黄鱼(Larimichthyscrocea)肌肉脂肪酸组成的营养学方法。
背景技术
大黄鱼属于硬骨鱼纲的鲈形目、石首鱼科、黄鱼属，主要分布在我国东南沿海海域，其肉质鲜美，且具有一定的药用价值，是我国特有海水经济鱼类。根据2021年中国渔业统计年鉴显示，大黄鱼的养殖产量超过25万吨，是我国养殖产量最大的海水经济鱼类。
近年来，高脂饲料被广泛应用于大黄鱼饲料中，因为许多研究表明，高脂饲料可以促进鱼类生长，提高蛋白质的利用率，降低水产养殖成本。但是，长期使用高脂饲料会导致鱼体脂质代谢紊乱，降低鱼肉的营养价值。因此，寻找有效的策略来缓解高脂饲料带来的不利影响刻不容缓。
肠道菌群被称为机体的第二大基因组，对宿主的消化吸收和营养代谢等生理过程具有重要的影响。研究发现，肠道中厚壁菌门和拟杆菌的比例与机体的肥胖程度密切相关。此外，肠道菌群的代谢产物如丁酸、胆汁酸和色氨酸等也会对机体的脂肪代谢产生较大的影响。因此，探究肠道菌群对大黄鱼肌肉脂肪酸组成的调控作用，有望缓解水产养殖中使用高脂饲料引发鱼肉质量下降的问题。
发明内容
本发明要解决的问题在于提供一种通过肠道菌群调控高脂投喂下大黄鱼肌肉脂肪酸组成的营养学方法，所述方法聚集1种特定饲料功能性物质，通过添加其在饲料中特定比例来改变大黄鱼肠道菌群的组成，进而达到调控大黄鱼肌肉脂肪酸组成的问题，以期解决高脂饲料的负面影响。
本发明是通过如下技术方案来实现的：
一种通过肠道菌群调控高脂投喂下大黄鱼肌肉脂肪酸组成的营养学方法，它的方法如下：在大黄鱼高脂饲料中添加含量为0.5％的植物甾醇。
本发明还提供一种含有植物甾醇的大黄鱼饲料，所述植物甾醇在大黄鱼饲料中的比例为0.5％。
本发明还提供植物甾醇在制备大黄鱼饲料添加剂中的应用。
本发明还提供一种含有植物甾醇的大黄鱼饲料添加剂。
本发明与现有技术相比的有益效果：
1)植物甾醇对鱼体无毒副作用，对食品安全无任何影响。本发明通过在高脂饲料中添加0.5％植物甾醇，能够有效缓解高脂饲料引发的肠道菌群紊乱，提高菌群丰富度、改变菌群结构；提高拟杆菌门和厚壁菌门的比例。
2)本发明通过在高脂大黄鱼饲料中添加0.5％的植物甾醇能够显著降低肌肉脂肪酸组成中SFA的含量比例，并在一定程度上提高n-6PUFA和n-3PUFA的含量比例，提高鱼肉的营养价值。
附图说明
图1、本发明实施例1-2中各组间大黄鱼肠道菌群OUT数及维恩图；NO为适宜组，HO为高脂组，PS0.5为添加组；
图2、本发明实施例1-2中各组间大黄鱼肠道菌群的α-多样性指标：A、Observedspecies指数箱形图；B、Shannon指数箱形图；C、Simpson指数箱形图；D、Chao1指数箱形图；NO为适宜组，HO为高脂组，PS0.5为添加组；
图3、本发明实施例1-2中各组间大黄鱼肠道菌群的β-多样性指标：A、主成分分析；B、UPGMA距离图；NO为适宜组，HO为高脂组，PS0.5为添加组。
图4、本发明实施例1-2中大黄鱼肠道菌群相对丰度图：A、门水平相对丰度图；B、属水平相对丰度图；C、种水平相对丰度图；NO为适宜组，HO为高脂组，PS0.5为添加组。
图5、本发明实施例1中大黄鱼肌肉脂肪酸组成比例图：NO为适宜组，HO为高脂组，PS0.5为添加组。
图6、本发明实施例1-2中大黄鱼肠道菌群组成与肌肉脂肪酸组成比例相关性分析热图。
具体实施方式
下面通过实施例对本发明的技术特征作进一步解释，但本发明的保护范围不受实施例任何形式上的限制。
实施例1、使用本方法进行饲料制造及养殖管理
1、实验设计和实验饲料配方
以本实验室往年所用的大黄鱼养殖高脂饲料组(18％脂肪水平)为阴性对照组，在阴性对照组中添加0.5％(即5mg/kg)的植物甾醇为添加组。另外，以适宜饲料组(12％脂肪水平)作为阳性对照组。饲料配方见表1。
表1实验饲料的配方和粗成分(％干成分)
1白鱼粉(74％粗蛋白，7.84％粗脂肪)；高级磷虾粉(64.86％粗蛋白，8％粗脂肪)；去皮豆粕(54.08％粗蛋白，0.35％粗脂肪)；高筋面粉(21.2％粗蛋白，0.34％粗脂肪)。
2多维预混料(mg/kg diet)(维生素A醋酸酯,32；维生素D3,5；α-生育酚,240；维生素B1,25；维生素B2,45；盐酸吡哆醇,20；维生素B12,10；泛酸,60；叶酸,20；烟酸,200；生物素,60；肌醇,800；维晶纤维素,13473)。
3多矿预混料(mg/kg diet)(硫酸镁，1200；硫酸铜，10；硫酸亚铁，80，硫酸锌，50；硫酸锰，45；氯化钴，50；亚硒酸钠，20；碘酸钙，60；沸石粉，13485)。
4甘氨酸和甜菜碱。
5植物甾醇(42.3％β-谷甾醇、29.1％豆甾醇和23.9％菜油甾醇)。
2、实验用鱼和养殖管理
本实验选用初始体重为13.08±0.02g的实验大黄鱼幼鱼共540尾，鱼苗购自宁德市富发水产有限公司。随机分成三个组，每个组三个重复，每个重复60尾鱼，养殖周期为70天。在正式养殖实验前，实验幼鱼被放置在4m×4m×4m的网箱中，投喂商业饲料进行为期两周的暂养。正式实验开始后，每天5:30和17:30进行饱食投喂，不定期清理水面杂物、打捞死鱼。实验期间水温维持在19.3-22.8℃，盐度在25.6-29.9‰，溶氧水平在6.1-7.0mg/L之间。
3、实验样品采集与分析
养殖实验结束后，在经过24小时的禁食以后进行取样。每网取18尾鱼的肌肉样品，混合后立即放入-80℃液氮中保存，待进一步分析。同时，另取3尾鱼，点燃酒精灯做无菌环境，使用酒精对取样器材消毒并擦拭鱼体表面，随后解剖鱼体取肠，轻刮肠壁并收集，立即放入-80℃液氮中保存。用于肠道菌群分析。
将肌肉组织在冻干室中冷冻干燥，使用HP6890气相色谱仪、毛细管柱和火焰离子化检测器识别和定量脂肪酸甲酯，用于计算肌肉脂肪酸组成。
如图1显示，经过大黄鱼肠道菌群高通测序后，经聚类发现适宜组、高脂组和添加组共得到1269个OTUs(97％相似度)，且三个处理组共有233个OTUs。
适宜组得到699个OUTs，高脂组得到720个OTUs，添加组得到695个OTUs，其中适宜组、高脂组和添加组独有的分别为255、179和223个OTUs。结果表明，相比较适宜组，高脂组肠道微生物OTUs数量上升，而添加组可降低肠道微生物中的OTUs数量。
如图2显示，肠道菌群的α-多样性结果表明，与适宜组相比，高脂组的Observedspecies指数和Shannon指数下降，而添加组的Observed species指数、Shannon指数、Simpson指数和Chao1指数得以提高，表明植物甾醇可以提高肠道微生物的α-多样性。
如图3所示，根据β-多样性分析中的主坐标分析(PCoA)结果可知，三组的样本聚集较近。UPGMA聚类结果显示，适宜组和添加组的聚类更相似且有更高分支，而高脂组与适宜组相距较远且形成单独分支。
如图4所示，适宜组、高脂组和添加组在门水平上的优势菌主要为变形菌门、放线菌门、厚壁菌门、蓝藻细菌门、拟杆菌门等。在属水平上优势菌主要为甲基杆菌属、劳尔氏菌属、水栖菌属、葡萄球菌属、嗜氢菌属等。在种水平上优势菌主要为皮氏罗尔斯顿菌、奥斯陆莫拉氏菌和Hydrogenophilus thermoluteolus等。在门、属和种水平上，各组间的优势菌相似，但组成比例有较大差异。与适宜组相比，高脂组的厚壁菌门的数量明显升高而拟杆菌门数量明显下降。当添加植物甾醇后，厚壁菌门的数量明显下降而拟杆菌门数量明显上升。
如图5所示，与适宜组和高脂组相比，添加组的肌肉SFA含量比例显著降低。而与高脂组相比，添加组的肌肉n-6PUFA和n-3PUFA含量比例有所上升。
如图6所示，通过斯皮尔曼相关性分析，大黄鱼肠道菌群在门和科水平上的改变与肌肉脂肪酸组成呈现显著正相关性或负相关性。表明本方法通过改变肠道菌群的组成进而调控肌肉脂肪酸组成。
Claims (4)
1.一种调控高脂投喂下大黄鱼肌肉脂肪酸组成的营养学方法，其特征在于所述方法如下：在大黄鱼高脂饲料中添加含量为0.5％的植物甾醇。
2.一种含有植物甾醇的大黄鱼饲料，其特征在于所述植物甾醇在大黄鱼饲料中的质量比为0.5％。
3.植物甾醇在大黄鱼饲料添加剂中的应用。
4.一种含有植物甾醇的大黄鱼饲料添加剂。
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