CN106701583B - 一种海水小球藻保存方法 - Google Patents

一种海水小球藻保存方法 Download PDF

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CN106701583B
CN106701583B CN201611245398.0A CN201611245398A CN106701583B CN 106701583 B CN106701583 B CN 106701583B CN 201611245398 A CN201611245398 A CN 201611245398A CN 106701583 B CN106701583 B CN 106701583B
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云天颖
袁卫
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Hainan Green Algae World Biotechnology Co ltd
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Abstract

本发明提供了一种海水小球藻保存方法,能满足高浓度海水小球藻在常温(20℃‑25℃)避光条件下长时间保存,该保存方法经济环保、操作便捷,该方法包含如下步骤:(1)配制保鲜液,每份所述保鲜液的组份及用量如下:R‑多糖1‑8g,102‑103cfu/ml复合芽孢杆菌1‑7ml,102‑103cfu/ml亚硝化菌1‑7ml,102‑103cfu/ml硝化细菌1‑7ml,101‑102cfu/ml乳酸菌1‑7ml,NH2CONH2 0.1‑0.5g,KH2PO4 0.1‑0.4g;(2)将需要保存的小球藻液与所述保鲜液均匀混合,1升小球藻液使用1份保鲜液,装入塑料桶中、密封保存。

Description

一种海水小球藻保存方法
技术领域
本发明涉及微藻养殖技术领域,具体涉及一种海水小球藻保存方法。
背景技术
小球藻细胞中含有多种高价值的营养成分和化工原料,细胞代谢产生的多糖、蛋白质、色素等,在医药工业、食品工业、动物饲料、环境检测及净化、生物技术以及可再生能源制造等方面发挥着越来越重要的作用。目前,小球藻在水产养殖方面的应用日益重要,海水小球藻由于藻体内EPA、DHA含量比淡水小球藻高,在轮虫和贝类饵料方面应用效果更加突出,并在鱼虾、贝类育苗养殖中得到广泛应用;然而,天然海水中小球藻数量太少,满足不了高密度培养的鱼虾幼体、贝类幼体最佳生长的需要,所以需要供足够量的人工培养、经过优选、具有高营养价值的小球藻。
但是,目前海水小球藻浓缩液保存技术较为落后,主要表现在(1)高浓度藻液受生存空间影响,导致藻体不适,易被其他病源微生物侵染,导致腐败变质,进而影响饵料食用效果;(2)保存条件要求严苛,需在低温冷藏条件下进行保存,保存成本高;(3)目前较好的保鲜液由于价钱昂贵而不被市场接受。
发明内容
针对上述出现的问题,本发明提供了一种海水小球藻保存方法,能满足高浓度海水小球藻在常温(20℃-25℃)避光阴凉条件下长时间保存,该保存方法经济环保、操作便捷,该方法包含如下步骤:
(1)配制保鲜液,每份所述保鲜液的组份及用量如下:
R-多糖1-8g,102-103cfu/ml复合芽孢杆菌1-7ml,102-103cfu/ml亚硝化菌1-7ml,102-103cfu/ml硝化细菌1-7ml,101-102cfu/ml乳酸菌1-7ml,NH2CONH2 0.1-0.5g,KH2PO40.1-0.4g;
进一步的,每份所述保鲜液的优选配方为:R-多糖2-5g,102-103cfu/ml复合芽孢杆菌2-4ml,102-103cfu/ml亚硝化菌2-4ml,102-103cfu/ml硝化细菌2-4ml,101-102cfu/ml乳酸菌2-4ml,NH2CONH2 0.1-0.3g,KH2PO4 0.2-0.3g;
进一步的,每份所述保鲜液的优选配方为:R-多糖5g,102cfu/ml复合芽孢杆菌2ml,102cfu/ml亚硝化菌2ml,103cfu/ml硝化细菌2ml,10cfu/ml乳酸菌2ml,NH2CONH20.13g,KH2PO4 0.17g;
(2)将需要保存的小球藻液与所述保鲜液均匀混合,1升小球藻液使用1份保鲜液,装入塑料桶中、密封保存。
本发明实现了小球藻浓度液(10×108cell)在常温(20℃-25℃)、避光阴凉条件下保存30天,小球藻生物量占初始生物量90%以上,藻液颜色保持翠绿色,藻液气味正常。
具体实施方式
实施例一
1.将培养得到的海水小球藻液采收离心后收集藻泥,用灭菌海水稀释小球藻液浓度至10×108cell;
2.配制保鲜液,保鲜液配方如下:
组份 用量
R-多糖 1g
10<sup>2</sup>cfu复合芽孢杆菌 1ml
10<sup>2</sup>cfu亚硝化菌 1ml
10<sup>3</sup>cfu硝化细菌 1ml
10cfu乳酸菌 1ml
NH<sub>2</sub>CONH<sub>2</sub> 0.1g
KH<sub>2</sub>PO<sub>4</sub> 0.1g
3.将1L小球藻浓度液与上述保鲜液均匀混合后,装入塑料桶中,并密封,于常温25℃、避光阴凉条件下放置观察,同样方法制备相同规格的6份样品。
4.分别在第25日、第26日、第27日、第28日、第29日、第30日随机选取上述6个样品中的一个开封,并进行显微镜计数,观察颜色变化情况,藻液气味变化情况,结果如表1所示。
实施例二
1.将培养得到的海水小球藻液采收离心后收集藻泥,用灭菌海水稀释小球藻液浓度至10×108cell;
2.配制保鲜液,保鲜液配方如下:
Figure BDA0001196972010000021
Figure BDA0001196972010000031
3.将1L小球藻浓度液与上述保鲜液均匀混合后,装入塑料桶中,并密封,于常温25℃、避光阴凉条件下放置观察,同样方法制备相同规格的6份样品。
4.分别在第25日、第26日、第27日、第28日、第29日、第30日随机选取上述6个样品中的任一个开封,并进行显微镜计数,观察颜色变化情况,藻液气味变化情况,结果如表1所示。
实施例三
1.将培养得到的海水小球藻液采收离心后收集藻泥,用灭菌海水稀释小球藻液浓度至10×108cell;
2.配制保鲜液,保鲜液配方如下:
组份 用量
R-多糖 8g
10<sup>2</sup>cfu复合芽孢杆菌 7ml
10<sup>2</sup>cfu亚硝化菌 7ml
10<sup>3</sup>cfu硝化细菌 7ml
10cfu乳酸菌 7ml
NH<sub>2</sub>CONH<sub>2</sub> 0.5g
KH<sub>2</sub>PO<sub>4</sub> 0.4g
3.将1L小球藻浓度液与上述保鲜液均匀混合后,装入塑料桶中,并密封,于常温25℃、避光阴凉条件下放置观察,同样方法制备相同规格的6份样品。
4.分别在第25日、第26日、第27日、第28日、第29日、第30日随机选取上述6个样品中的任一个开封,并进行显微镜计数,观察颜色变化情况,藻液气味变化情况,结果如表1所示。
表1小球藻液活细胞数量、气味及颜色变化情况
Figure BDA0001196972010000032
Figure BDA0001196972010000041

Claims (3)

1.一种海水小球藻保存方法,其特征在于,该保存方法包括如下步骤:
(1)配制保鲜液,每份所述保鲜液的组份及用量如下:
R-多糖1-8g,102-103cfu/ml复合芽孢杆菌1-7ml,102-103cfu/ml亚硝化菌1-7ml,102-103cfu/ml硝化细菌1-7ml,101-102cfu/ml乳酸菌1-7ml,NH2CONH2 0.1-0.5g,KH2PO4 0.1-0.4g;
(2)将需要保存的小球藻液与所述保鲜液均匀混合,1升小球藻液使用1份保鲜液,装入塑料桶中、密封保存;
所述小球藻液与保鲜液混合后,在20℃-25℃、避光、阴凉条件下保存。
2.如权利要求1所述的海水小球藻保存方法,其特征在于,每份所述保鲜液的配方为:R-多糖2-5g,102-103cfu/ml复合芽孢杆菌2-4ml,102-103cfu/ml亚硝化菌2-4ml,102-103cfu/ml硝化细菌2-4ml,101-102cfu/ml乳酸菌2-4ml,NH2CONH2 0.1-0.3g,KH2PO4 0.2-0.3g。
3.如权利要求2所述的海水小球藻保存方法,其特征在于,每份所述保鲜液的配方为:R-多糖5g,102cfu/ml复合芽孢杆菌2ml,102cfu/ml亚硝化菌2ml,103cfu/ml硝化细菌2ml,10cfu/ml乳酸菌2ml,NH2CONH2 0.13g,KH2PO4 0.17g。
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