CN106701583A - 一种海水小球藻保存方法 - Google Patents
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- 241000195649 Chlorella <Chlorellales> Species 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 19
- 239000013535 sea water Substances 0.000 title abstract description 8
- 238000004321 preservation Methods 0.000 title abstract description 5
- 239000007788 liquid Substances 0.000 claims abstract description 36
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 18
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- 229920001282 polysaccharide Polymers 0.000 claims abstract description 10
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- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 9
- 239000002131 composite material Substances 0.000 claims abstract description 8
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims abstract description 5
- 241000195493 Cryptophyta Species 0.000 claims description 23
- 239000011324 bead Substances 0.000 claims description 14
- 241001495402 Nitrococcus Species 0.000 claims description 8
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 7
- 238000007789 sealing Methods 0.000 claims description 2
- 244000063299 Bacillus subtilis Species 0.000 abstract 1
- 235000014469 Bacillus subtilis Nutrition 0.000 abstract 1
- 235000019796 monopotassium phosphate Nutrition 0.000 abstract 1
- 238000005119 centrifugation Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000003306 harvesting Methods 0.000 description 3
- 235000015170 shellfish Nutrition 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 241000143060 Americamysis bahia Species 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 241000726221 Gemma Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000700141 Rotifera Species 0.000 description 1
- 239000000877 Sex Attractant Substances 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- NNBFNNNWANBMTI-UHFFFAOYSA-M brilliant green Chemical compound OS([O-])(=O)=O.C1=CC(N(CC)CC)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](CC)CC)C=C1 NNBFNNNWANBMTI-UHFFFAOYSA-M 0.000 description 1
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- 150000004676 glycans Chemical class 0.000 description 1
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- 239000004615 ingredient Substances 0.000 description 1
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- 235000016709 nutrition Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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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
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 |
102cfu复合芽孢杆菌 | 1ml |
102cfu亚硝化菌 | 1ml |
103cfu硝化细菌 | 1ml |
10cfu乳酸菌 | 1ml |
NH2CONH2 | 0.1g |
KH2PO4 | 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:
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 |
102cfu复合芽孢杆菌 | 7ml |
102cfu亚硝化菌 | 7ml |
103cfu硝化细菌 | 7ml |
10cfu乳酸菌 | 7ml |
NH2CONH2 | 0.5g |
KH2PO4 | 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
Claims (4)
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,KH2PO40.1-0.4g;
(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。
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。
4.如权利要求1所述的海水小球藻保存方法,其特征在于,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。
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CN108342324A (zh) * | 2018-04-10 | 2018-07-31 | 中盐工程技术研究院有限公司 | 一种小球藻保存方法 |
CN108753623A (zh) * | 2018-06-12 | 2018-11-06 | 兰溪市沉默生物科技有限公司 | 一种小球藻保存方法 |
CN113278557A (zh) * | 2021-05-25 | 2021-08-20 | 海南绿藻世界生物科技有限公司 | 共生菌组合物及其制备方法和微藻的培养方法 |
EP3933024A1 (en) * | 2020-07-02 | 2022-01-05 | LG Chem, Ltd. | Gas collection device |
CN114686392A (zh) * | 2020-12-31 | 2022-07-01 | 中国石油化工股份有限公司 | 一种硝化细菌的常温保藏方法 |
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Publication number | Priority date | Publication date | Assignee | Title |
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EP3933024A1 (en) * | 2020-07-02 | 2022-01-05 | LG Chem, Ltd. | Gas collection device |
CN114686392A (zh) * | 2020-12-31 | 2022-07-01 | 中国石油化工股份有限公司 | 一种硝化细菌的常温保藏方法 |
CN114686392B (zh) * | 2020-12-31 | 2023-07-04 | 中国石油化工股份有限公司 | 一种硝化细菌的常温保藏方法 |
CN113278557A (zh) * | 2021-05-25 | 2021-08-20 | 海南绿藻世界生物科技有限公司 | 共生菌组合物及其制备方法和微藻的培养方法 |
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