CN113678942B - 小球藻的应用 - Google Patents
小球藻的应用 Download PDFInfo
- Publication number
- CN113678942B CN113678942B CN202110890806.2A CN202110890806A CN113678942B CN 113678942 B CN113678942 B CN 113678942B CN 202110890806 A CN202110890806 A CN 202110890806A CN 113678942 B CN113678942 B CN 113678942B
- Authority
- CN
- China
- Prior art keywords
- chlorella
- powder
- acid
- fish
- feed
- 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.)
- Active
Links
- 241000195649 Chlorella <Chlorellales> Species 0.000 title claims abstract description 106
- 239000000843 powder Substances 0.000 claims abstract description 54
- 241000195651 Chlorella sp. Species 0.000 claims abstract description 23
- 239000003674 animal food additive Substances 0.000 claims abstract description 22
- 239000001963 growth medium Substances 0.000 claims abstract description 12
- 238000004321 preservation Methods 0.000 claims abstract description 5
- 239000000203 mixture Substances 0.000 claims description 15
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 7
- 229940041514 candida albicans extract Drugs 0.000 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- 239000012138 yeast extract Substances 0.000 claims description 7
- 238000005286 illumination Methods 0.000 claims description 6
- 239000002609 medium Substances 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 4
- 238000001471 micro-filtration Methods 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 2
- 239000011148 porous material Substances 0.000 claims description 2
- 239000012465 retentate Substances 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims 1
- 241000251468 Actinopterygii Species 0.000 abstract description 28
- 241000195493 Cryptophyta Species 0.000 abstract description 26
- 235000016709 nutrition Nutrition 0.000 abstract description 12
- 230000000968 intestinal effect Effects 0.000 abstract description 11
- 238000002474 experimental method Methods 0.000 abstract description 9
- 230000012010 growth Effects 0.000 abstract description 7
- 230000036541 health Effects 0.000 abstract description 6
- 239000004615 ingredient Substances 0.000 abstract description 4
- 241000276707 Tilapia Species 0.000 description 20
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 20
- 238000012360 testing method Methods 0.000 description 16
- DTOSIQBPPRVQHS-PDBXOOCHSA-N alpha-linolenic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(O)=O DTOSIQBPPRVQHS-PDBXOOCHSA-N 0.000 description 12
- 238000004458 analytical method Methods 0.000 description 12
- 238000012163 sequencing technique Methods 0.000 description 12
- 238000000034 method Methods 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 241000894007 species Species 0.000 description 8
- TUNFSRHWOTWDNC-UHFFFAOYSA-N Myristic acid Natural products CCCCCCCCCCCCCC(O)=O TUNFSRHWOTWDNC-UHFFFAOYSA-N 0.000 description 7
- 235000020661 alpha-linolenic acid Nutrition 0.000 description 7
- 230000037396 body weight Effects 0.000 description 7
- POULHZVOKOAJMA-UHFFFAOYSA-N dodecanoic acid Chemical compound CCCCCCCCCCCC(O)=O POULHZVOKOAJMA-UHFFFAOYSA-N 0.000 description 7
- 238000002156 mixing Methods 0.000 description 7
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 6
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 6
- 239000002253 acid Substances 0.000 description 6
- 235000014113 dietary fatty acids Nutrition 0.000 description 6
- ZQPPMHVWECSIRJ-MDZDMXLPSA-N elaidic acid Chemical compound CCCCCCCC\C=C\CCCCCCCC(O)=O ZQPPMHVWECSIRJ-MDZDMXLPSA-N 0.000 description 6
- 229930195729 fatty acid Natural products 0.000 description 6
- 239000000194 fatty acid Substances 0.000 description 6
- 150000004665 fatty acids Chemical class 0.000 description 6
- 229960004488 linolenic acid Drugs 0.000 description 6
- 229930182817 methionine Natural products 0.000 description 6
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 6
- SECPZKHBENQXJG-FPLPWBNLSA-N palmitoleic acid Chemical compound CCCCCC\C=C/CCCCCCCC(O)=O SECPZKHBENQXJG-FPLPWBNLSA-N 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 239000002994 raw material Substances 0.000 description 6
- 108020004414 DNA Proteins 0.000 description 5
- 235000021314 Palmitic acid Nutrition 0.000 description 5
- 235000001014 amino acid Nutrition 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 description 4
- JEBFVOLFMLUKLF-IFPLVEIFSA-N Astaxanthin Natural products CC(=C/C=C/C(=C/C=C/C1=C(C)C(=O)C(O)CC1(C)C)/C)C=CC=C(/C)C=CC=C(/C)C=CC2=C(C)C(=O)C(O)CC2(C)C JEBFVOLFMLUKLF-IFPLVEIFSA-N 0.000 description 4
- 244000249214 Chlorella pyrenoidosa Species 0.000 description 4
- 235000007091 Chlorella pyrenoidosa Nutrition 0.000 description 4
- 241000195628 Chlorophyta Species 0.000 description 4
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 4
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 4
- 239000004472 Lysine Substances 0.000 description 4
- 229930182558 Sterol Natural products 0.000 description 4
- 238000007605 air drying Methods 0.000 description 4
- OENHQHLEOONYIE-UKMVMLAPSA-N all-trans beta-carotene Natural products CC=1CCCC(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C OENHQHLEOONYIE-UKMVMLAPSA-N 0.000 description 4
- 235000013793 astaxanthin Nutrition 0.000 description 4
- 239000001168 astaxanthin Substances 0.000 description 4
- MQZIGYBFDRPAKN-ZWAPEEGVSA-N astaxanthin Chemical compound C([C@H](O)C(=O)C=1C)C(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)C(=O)[C@@H](O)CC1(C)C MQZIGYBFDRPAKN-ZWAPEEGVSA-N 0.000 description 4
- 229940022405 astaxanthin Drugs 0.000 description 4
- 235000013734 beta-carotene Nutrition 0.000 description 4
- 239000011648 beta-carotene Substances 0.000 description 4
- TUPZEYHYWIEDIH-WAIFQNFQSA-N beta-carotene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2=CCCCC2(C)C TUPZEYHYWIEDIH-WAIFQNFQSA-N 0.000 description 4
- 229960002747 betacarotene Drugs 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000003797 essential amino acid Substances 0.000 description 4
- 235000020776 essential amino acid Nutrition 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- VKOBVWXKNCXXDE-UHFFFAOYSA-N icosanoic acid Chemical compound CCCCCCCCCCCCCCCCCCCC(O)=O VKOBVWXKNCXXDE-UHFFFAOYSA-N 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 239000003921 oil Substances 0.000 description 4
- 235000019198 oils Nutrition 0.000 description 4
- 150000003432 sterols Chemical class 0.000 description 4
- 235000003702 sterols Nutrition 0.000 description 4
- OENHQHLEOONYIE-JLTXGRSLSA-N β-Carotene Chemical compound CC=1CCCC(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C OENHQHLEOONYIE-JLTXGRSLSA-N 0.000 description 4
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 3
- TWJNQYPJQDRXPH-UHFFFAOYSA-N 2-cyanobenzohydrazide Chemical compound NNC(=O)C1=CC=CC=C1C#N TWJNQYPJQDRXPH-UHFFFAOYSA-N 0.000 description 3
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 3
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 3
- 235000017060 Arachis glabrata Nutrition 0.000 description 3
- 244000105624 Arachis hypogaea Species 0.000 description 3
- 235000010777 Arachis hypogaea Nutrition 0.000 description 3
- 235000018262 Arachis monticola Nutrition 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 239000005639 Lauric acid Substances 0.000 description 3
- 235000021360 Myristic acid Nutrition 0.000 description 3
- 239000005642 Oleic acid Substances 0.000 description 3
- 235000021319 Palmitoleic acid Nutrition 0.000 description 3
- 235000021355 Stearic acid Nutrition 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- SECPZKHBENQXJG-UHFFFAOYSA-N cis-palmitoleic acid Natural products CCCCCCC=CCCCCCCCC(O)=O SECPZKHBENQXJG-UHFFFAOYSA-N 0.000 description 3
- 230000002354 daily effect Effects 0.000 description 3
- 235000019197 fats Nutrition 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 3
- 235000020778 linoleic acid Nutrition 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 3
- 235000020660 omega-3 fatty acid Nutrition 0.000 description 3
- 235000020232 peanut Nutrition 0.000 description 3
- 150000003904 phospholipids Chemical class 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 239000005017 polysaccharide Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000013535 sea water Substances 0.000 description 3
- 239000003549 soybean oil Substances 0.000 description 3
- 235000012424 soybean oil Nutrition 0.000 description 3
- 239000008117 stearic acid Substances 0.000 description 3
- DVSZKTAMJJTWFG-SKCDLICFSA-N (2e,4e,6e,8e,10e,12e)-docosa-2,4,6,8,10,12-hexaenoic acid Chemical compound CCCCCCCCC\C=C\C=C\C=C\C=C\C=C\C=C\C(O)=O DVSZKTAMJJTWFG-SKCDLICFSA-N 0.000 description 2
- MJYQFWSXKFLTAY-OVEQLNGDSA-N (2r,3r)-2,3-bis[(4-hydroxy-3-methoxyphenyl)methyl]butane-1,4-diol;(2r,3r,4s,5s,6r)-6-(hydroxymethyl)oxane-2,3,4,5-tetrol Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O.C1=C(O)C(OC)=CC(C[C@@H](CO)[C@H](CO)CC=2C=C(OC)C(O)=CC=2)=C1 MJYQFWSXKFLTAY-OVEQLNGDSA-N 0.000 description 2
- GZJLLYHBALOKEX-UHFFFAOYSA-N 6-Ketone, O18-Me-Ussuriedine Natural products CC=CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O GZJLLYHBALOKEX-UHFFFAOYSA-N 0.000 description 2
- 241000604933 Bdellovibrio Species 0.000 description 2
- 240000004082 Bidens tripartita Species 0.000 description 2
- 235000000621 Bidens tripartita Nutrition 0.000 description 2
- 241000186000 Bifidobacterium Species 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241000195654 Chlorella sorokiniana Species 0.000 description 2
- 241000196319 Chlorophyceae Species 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 2
- 241000168517 Haematococcus lacustris Species 0.000 description 2
- 108091023242 Internal transcribed spacer Proteins 0.000 description 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 2
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 2
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 2
- 235000019687 Lamb Nutrition 0.000 description 2
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 2
- 235000021353 Lignoceric acid Nutrition 0.000 description 2
- CQXMAMUUWHYSIY-UHFFFAOYSA-N Lignoceric acid Natural products CCCCCCCCCCCCCCCCCCCCCCCC(=O)OCCC1=CC=C(O)C=C1 CQXMAMUUWHYSIY-UHFFFAOYSA-N 0.000 description 2
- 241000586744 Micractinium pusillum Species 0.000 description 2
- 241000195650 Oocystaceae Species 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 235000019764 Soybean Meal Nutrition 0.000 description 2
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 2
- 239000004473 Threonine Substances 0.000 description 2
- 235000004279 alanine Nutrition 0.000 description 2
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 235000011089 carbon dioxide Nutrition 0.000 description 2
- 238000012937 correction Methods 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 2
- KAUVQQXNCKESLC-UHFFFAOYSA-N docosahexaenoic acid (DHA) Natural products COC(=O)C(C)NOCC1=CC=CC=C1 KAUVQQXNCKESLC-UHFFFAOYSA-N 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000010201 enrichment analysis Methods 0.000 description 2
- FARYTWBWLZAXNK-WAYWQWQTSA-N ethyl (z)-3-(methylamino)but-2-enoate Chemical compound CCOC(=O)\C=C(\C)NC FARYTWBWLZAXNK-WAYWQWQTSA-N 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 238000011049 filling Methods 0.000 description 2
- 235000004426 flaxseed Nutrition 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 208000006637 fused teeth Diseases 0.000 description 2
- VZCCETWTMQHEPK-UHFFFAOYSA-N gamma-Linolensaeure Natural products CCCCCC=CCC=CCC=CCCCCC(O)=O VZCCETWTMQHEPK-UHFFFAOYSA-N 0.000 description 2
- VZCCETWTMQHEPK-QNEBEIHSSA-N gamma-linolenic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/CCCCC(O)=O VZCCETWTMQHEPK-QNEBEIHSSA-N 0.000 description 2
- 235000020664 gamma-linolenic acid Nutrition 0.000 description 2
- 229960002733 gamolenic acid Drugs 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 210000000514 hepatopancreas Anatomy 0.000 description 2
- 239000012510 hollow fiber Substances 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- 229940012843 omega-3 fatty acid Drugs 0.000 description 2
- 239000006014 omega-3 oil Substances 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 238000006213 oxygenation reaction Methods 0.000 description 2
- 210000000496 pancreas Anatomy 0.000 description 2
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 2
- 244000144977 poultry Species 0.000 description 2
- 235000013594 poultry meat Nutrition 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 230000003134 recirculating effect Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 238000007873 sieving Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000004455 soybean meal Substances 0.000 description 2
- QZZGJDVWLFXDLK-UHFFFAOYSA-N tetracosanoic acid Chemical compound CCCCCCCCCCCCCCCCCCCCCCCC(O)=O QZZGJDVWLFXDLK-UHFFFAOYSA-N 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- JBYXPOFIGCOSSB-GOJKSUSPSA-N 9-cis,11-trans-octadecadienoic acid Chemical compound CCCCCC\C=C\C=C/CCCCCCCC(O)=O JBYXPOFIGCOSSB-GOJKSUSPSA-N 0.000 description 1
- 244000153158 Ammi visnaga Species 0.000 description 1
- 235000010585 Ammi visnaga Nutrition 0.000 description 1
- 240000002900 Arthrospira platensis Species 0.000 description 1
- 235000016425 Arthrospira platensis Nutrition 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
- 240000009108 Chlorella vulgaris Species 0.000 description 1
- 235000007089 Chlorella vulgaris Nutrition 0.000 description 1
- 241000194033 Enterococcus Species 0.000 description 1
- 235000019733 Fish meal Nutrition 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 238000007696 Kjeldahl method Methods 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000276701 Oreochromis mossambicus Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 241000283903 Ovis aries Species 0.000 description 1
- 244000289453 Parkinsonia aculeata Species 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 241000195648 Pseudochlorella pringsheimii Species 0.000 description 1
- 239000013614 RNA sample Substances 0.000 description 1
- 238000000944 Soxhlet extraction Methods 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 235000006468 Thea sinensis Nutrition 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 241001293481 Trebouxiophyceae Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- MBLBDJOUHNCFQT-LXGUWJNJSA-N aldehydo-N-acetyl-D-glucosamine Chemical compound CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000000035 biogenic effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- YYRMJZQKEFZXMX-UHFFFAOYSA-L calcium bis(dihydrogenphosphate) Chemical compound [Ca+2].OP(O)([O-])=O.OP(O)([O-])=O YYRMJZQKEFZXMX-UHFFFAOYSA-L 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 230000035425 carbon utilization Effects 0.000 description 1
- 235000021466 carotenoid Nutrition 0.000 description 1
- 150000001747 carotenoids Chemical class 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 206010008118 cerebral infarction Diseases 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- ZXFCRFYULUUSDW-OWXODZSWSA-N chembl2104970 Chemical compound C([C@H]1C2)C3=CC=CC(O)=C3C(=O)C1=C(O)[C@@]1(O)[C@@H]2CC(O)=C(C(=O)N)C1=O ZXFCRFYULUUSDW-OWXODZSWSA-N 0.000 description 1
- 235000013330 chicken meat Nutrition 0.000 description 1
- 235000019804 chlorophyll Nutrition 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 229940108924 conjugated linoleic acid Drugs 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 235000019784 crude fat Nutrition 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 210000002969 egg yolk Anatomy 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 230000006862 enzymatic digestion Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000013100 final test Methods 0.000 description 1
- 239000004467 fishmeal Substances 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 210000003736 gastrointestinal content Anatomy 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 150000003278 haem Chemical class 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000012165 high-throughput sequencing Methods 0.000 description 1
- 230000034435 immune system development Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- -1 iron ion Chemical class 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- KQQKGWQCNNTQJW-UHFFFAOYSA-N linolenic acid Natural products CC=CCCC=CCC=CCCCCCCCC(O)=O KQQKGWQCNNTQJW-UHFFFAOYSA-N 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 235000012680 lutein Nutrition 0.000 description 1
- 239000001656 lutein Substances 0.000 description 1
- 229960005375 lutein Drugs 0.000 description 1
- KBPHJBAIARWVSC-RGZFRNHPSA-N lutein Chemical compound C([C@H](O)CC=1C)C(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\[C@H]1C(C)=C[C@H](O)CC1(C)C KBPHJBAIARWVSC-RGZFRNHPSA-N 0.000 description 1
- ORAKUVXRZWMARG-WZLJTJAWSA-N lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C ORAKUVXRZWMARG-WZLJTJAWSA-N 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000002366 mineral element Substances 0.000 description 1
- 229910000150 monocalcium phosphate Inorganic materials 0.000 description 1
- 235000019691 monocalcium phosphate Nutrition 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000003001 serine protease inhibitor Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 229940082787 spirulina Drugs 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- KBPHJBAIARWVSC-XQIHNALSSA-N trans-lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C KBPHJBAIARWVSC-XQIHNALSSA-N 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- FJHBOVDFOQMZRV-XQIHNALSSA-N xanthophyll Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C=C(C)C(O)CC2(C)C FJHBOVDFOQMZRV-XQIHNALSSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
- C12N1/125—Unicellular algae isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/89—Algae ; Processes using algae
-
- 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
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Landscapes
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Fodder In General (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一种小球藻GB‑116(Chlorella sp.GB‑116)的应用，该小球藻已于2020年8月13日保存于中国典型培养物保藏中心(简称CCTCC，地址：湖北省武汉市武昌区八一路299号武汉大学校内)，其保藏编号为CCTCC No：M 2020373。申请人通过实验发现，小球藻GB‑116(Chlorella sp.GB‑116)或其经异养培养基培养后所得的小球藻藻粉提高鱼免疫指标和生长性能，并且改善鱼肠道菌群，可用于制备鱼类饲料添加剂；同时，藻粉中含有多种营养成分，可用于制备营养保健品。
Description
技术领域
本发明涉及小球藻，具体涉及小球藻在制备饲料添加剂、营养保健品中的应用。
背景技术
小球藻，单细胞真核生物，属绿藻门(Chlorophyta)、绿藻纲(Chlorophyceae)、绿球藻目(Chlorococcales)、卵囊藻科(Oocystaceae)、小球藻属(Chlorella)，直径2～12um，生长繁殖速度快。目前，我国主要有蛋白核小球藻、椭圆小球藻、普通小球藻这三种小球藻，人工养殖有自养、异养、兼养和混养等多种模式。
小球藻营养成分丰富，含有生长因子(GCF)、蛋白质、多种矿物质元素、多种氨基酸、维生素、以及不饱和脂肪酸等多种营养物质，小球藻的类胡萝卜素有β-胡萝卜素、叶黄素和虾青素。小球藻营养成分含量会因小球藻品种和培养方式不同而存在差异，魏文志等人通过在光自养模式下对7种小球藻进行营养成分分析，结果显示蛋白质含量在30.58～53.79％，其中蛋白核小球藻蛋白质含量最高，普通小球藻含量最低，而7种小球藻各氨基酸含量高低排名相似，都含有8种人体必需氨基酸，其含量在35.86～40.28％(魏文志,付立霞,&陈国宏.(2011).7种小球藻氨基酸组成及多元性分析.食品科学,32(5),254-257.)。小球藻中常见的脂肪酸组成为：C14:0(肉蔻酸)、C16:0(棕榈酸)、C16:1(棕榈油酸)、C18:0(硬脂酸)、C18:1(油酸)、C18:2(共轭亚麻油酸)、C18:3(亚麻酸)等，小球藻中含有36～43％多不饱和脂肪酸，其中亚油酸占多数，其他多不饱和脂肪酸则仅以少量或微量存在(Otles S,Pire R.Fatty acid composition of Chlorella and Spirulina microalgae species[J].Journal of AOAC International,2001,84(6):1708-1714.)。
近年来，小球藻作饲料添加剂广泛应用于畜牧、家禽、水产行业中，早在上个世纪60年代就有关于小球藻大面积生产后作家禽饲料的报道。在家鸡饲养中，日粮中添加亚麻籽和微藻可使蛋黄中α-亚麻酸(ALA)和二十二碳六烯酸(DHA)含量增加(Wu B,Dong X,WeiF,et al.Impact of feeding flaxseed and microalgal residue on omega-3fattyacids enrichment in hen eggs[J].Oil Crop Science,2017,2(4):265-271.)。在羔羊日粮中添加微藻后也有类似作用，可提高羔羊肌肉的二十二碳六烯酸(DHA)和总n3脂肪酸的含量(Diaz M T,Perez C,Sanchez C I,et al.Feeding microalgae increases omega3fatty acids of fat deposits and muscles in light lambs[J].Journal of FoodComposition and Analysis,2017,56:115-123.)。目前，研究人员对小球藻作为水产饲料添加剂其在生长性能、消化酶活性以及免疫等方面进行了研究，并取得了一定的进展，但小球藻对水产动物肠道微生物的影响少见研究，肠道微生物在促进宿主生长、代谢、免疫系统发育等方面起着重要作用。饲料添加剂对鱼类肠道微生物组成具有重要影响。研究小球藻作为水产饲料添加剂具有重要意义。
本申请发明人团队从海水中分离纯化得到了一种新的纯种小球藻，并对其进行了相关的研究。
发明内容
本发明要解决的技术问题是提供一种新的小球藻在制备饲料添加剂、营养保健品中的应用。
本发明的技术方案为：小球藻或小球藻藻粉在制备饲料添加剂或营养保健品中的应用，其中，
所述小球藻为小球藻GB-116(Chlorella sp.GB-116)，该藻株已于2020年8月13日保存于中国典型培养物保藏中心(简称CCTCC，地址：湖北省武汉市武昌区八一路299号武汉大学校内)，其保藏编号为CCTCC No：M 2020373；
所述小球藻藻粉是将小球藻GB-116(Chlorella sp.GB-116)置于异养培养基中，在23～26℃、光照强度为3500～4000lx的条件下进行培养；所得培养物经截留孔径为0.1～0.3μm的微滤膜过滤，收集截留物，干燥而得。
上述技术方案中，所述的小球藻GB-116(Chlorella sp.GB-116)从海水中分离得到。所述的异养培养基为改良的克氏培养基，其组成为：CaCl2·2H2O0.2～0.4g/L、FeSO4·7H2O 5～25mg/L、MgSO4·7H2O 0.05～0.2g/L、NaNO31.5～2.5g/L、K2HPO4 0.2～0.5g/L、酵母提取物0.5～1.5g/L、葡萄糖4～15g/L，pH＝6.0～8.0。所述培养通常在生物反应器中进行，转速优选设置为120～160r/min；在培养时优选为静置培养。所述的干燥通常是在不加热条件下进行，具体可以是在15～20℃条件下自然风干，或者是采用冷冻干燥。
上述技术方案中，所述的饲料添加剂具体可以是改善鱼肠道菌群的添加剂，或者是提高鱼免疫指标和生长性能的饲料添加剂。
申请人发现，小球藻GB-116(Chlorella sp.GB-116)藻株经过异养培养基培养后，该含有多种营养成分：其中总蛋白含量28.4％，多糖含量为1.29％，总脂肪含量1.1％，β-胡萝卜素含量127μg/g，虾青素含量0.850μg/g，总甾醇含量0.30％；17种人体必需氨基酸，其中蛋氨酸(甲硫氨酸)含量最高，达到36.205mg/g，其次是苏氨酸、丙氨酸、亮氨酸、赖氨酸、蛋氨酸和赖氨酸；15种脂肪酸：C12:0(月桂酸)、C14:0(肉蔻酸)、C15:0(十五碳酸)、C16:0(棕榈酸)、C16:1(棕榈油酸)、C17:0(十七碳酸)、C17:1(十七碳一烯酸)、C18:0(硬脂酸)、C18:1N9C(油酸)、C18:1N9T(反式油酸)、C18:2N6T(反式亚油酸)、C18:3N3(α-亚麻酸)、C18:3N6(γ-亚麻酸)、C20:0(花生酸)和C24:0(二十四碳酸)，其中C18:3N3(α-亚麻酸)是总油脂的主要成分，其含量仅次于最高含量的C16:0(棕榈酸)，为0.30552g/100g。本领域已知：C18:3N3(α-亚麻酸)具有多种生理功能，如增强智力，提高记忆力，保护视力、改善睡眠；抑制血栓性疾病，预防心肌梗死和脑梗死。降低血脂、降血压等，基于此，本发明提供小球藻或小球藻藻粉在制备营养保健品或饲料添加剂中的用途。
本发明还提供包含小球藻GB-116(Chlorella sp.GB-116)的细胞和/或其提取物和/或其培养物的食品组合物、营养补充剂或饲料添加剂。
申请人通过实验发现，小球藻GB-116(Chlorella sp.GB-116)或其经异养培养基培养后所得的小球藻藻粉提高鱼免疫指标和生长性能，并且改善鱼肠道菌群，可用于制备鱼类饲料添加剂；同时，藻粉中含有多种营养成分，可用于制备营养保健品。
附图说明
本发明涉及的小球藻GB-116(Chlorella sp.GB-116)，该藻株已于2020年8月13日保存于中国典型培养物保藏中心(简称CCTCC，地址：湖北省武汉市武昌区八一路299号武汉大学校内)，其保藏编号为CCTCC No：M 2020373。
图1为本发明所述小球藻GB-116(Chlorella sp.GB-116)在显微镜下镜检照片，放大倍数为40×10。
图2为本发明所述小球藻GB-116(Chlorella sp.GB-116)的ITS序列进化树。
图3为实施例2中对小球藻GB-116(Chlorella sp.GB-116)进行培养得到的小球藻藻粉17种水解氨基酸含量柱状图。
图4为实施例3中肝胰脏转录组测序分析流程图。
图5为实施例3中实验各分组在饲喂15天后体重的比较图。
图6为实施例4中扬道菌群16s rRNA测序分析流程图。
图7为实施例4中差异物种的LDA值分布柱状图。
具体实施方式
为了更好的解释本发明的技术方案，下面结合实施例及附图对本发明作进一步详细的描述，但本发明的实施方式不限于此。若未特别指明，实施例中所用的技术特征可以替换为具有在不背离发明构思前提下等同或相似功能或效果的其他本领域已知的技术特征。
实施例1：小球藻GB-116(Chlorella sp.GB-116)的分离和鉴定
1.小球藻的分离
将海水用无菌生理盐水稀释10倍、102倍、103倍、104倍、105倍后，各取200μL涂布于固体培养基(组成为：CaCl2·2H2O 1g/L、FeSO4·7H2O 0.01g/L、MgSO4·7H2O 0.2g/L、NaNO32g/L、K2HPO4 0.2g/L、酵母提取物1g/L、葡萄糖4g/L、琼脂1.5％，pH＝6.0～8.0)，23～26℃光照强度3750lx条件下培养3～5天，利用稀释平板涂布法分离得到单藻落，挑取单藻落再经多次平板划线进行纯化；之后用灭菌牙签挑取单菌落接种于5mL异养培养基得到一种纯种小球藻。
2.小球藻DNA提取
取对数生长期小球藻液10mL，8000r/min离心，用1.5mL无菌离心管收集小球藻，于液氮快速冻融3次，用无菌玻璃研磨棒快速研磨。小球藻DNA用CTAB植物基因组DNA快速提取试剂盒提取(详细步骤参考“北京艾德莱公司”试剂盒说明)。
3.小球藻ITS序列鉴定
用通用引物ITS1：5’-TCCGTAGGTGAACCTGCGG-3’(SEQ ID NO：1)和ITS4：5’-TCCTCCGCTTATTGATATGC-3’(SEQ ID NO：2)扩增真菌rDNA的间隔序列(含ITS1区、5.8S区、ITS2区)。配制50μL PCR扩增体系，PCR程序设置：94℃预变性3min；循环参数为：94℃变性30s，55℃退火30s，72度延伸1min，此为35个循环；之后72℃延伸10min。取3μL进行琼脂糖凝胶电泳检测，凝胶浓度为1％，电泳缓冲液为1×TAE缓冲液，将剩下PCR扩增产物送至苏州金唯智生物科技有限公司测序，所得ITS测序序列如下所示：
CGAAGTGAGAGCGTTCGGGGGGCCGAAACCACCACCGAATTCCTGCTAGGCCGCCAGCAAAGTCCCCTCGGGCCACGGCAGGCTGCGTAGCGGGTGCTACCTACCAAGCCATTGCCCTGCAAACGGGGTCCATGCTCAAGCCTCTACACTTCAGCCGACCCGGACGCAGCCTGTGACAGGCAACGGAGCCGGGAAGGCCAGGTCCGCACTCCCTCAGGGAGAGGGGGGCGAGGGTGTAAGCCGACGCTGAGGCAGACATGCCCTCGGCCGAAGCCTCGGGCGCAATTTGCGTTCAAAGATTCGATGGTTCACGGAATTCTGCAATTCACACTACGTATCGCATTTCGCTGCGTTCTTCATCGATACGGGAGCCAAGATATCCGTTGTTGAGAGTTGTCTTTGGTTTGTGCGGAGCGGACAGGCCGCCACCGCTGTTTGCTTCAGAGTGGTTTTGTGTTTGGGGTGGATTGAGTTGGATACAAATTACCAGCGTCACCAGCCGGCGAGGGCAAAGCCCAACGCCGGAGAGACGCCCGCCGGACCTGAAACTGCGGCTCGTAAGCCCCAGCCAGGGGACCGACGCCCGGCGGGAGCCGCCTTAGAAAAGGCAGGCCCAACCACACGGGGGGGACAAATGGTTACCAGGGTGGATTCGATCGATTCAATGATCCTTCCGCAGGTTCACCTCGGGTACTCCCACCAACCTGGTCGACAGGCTGGCTCCCTGGGCTCCGGGCTGGTTAGGCCGTTCCTCCCAGGGCACCCTCTCGGGTGGGCCGGACTCTATCTTAAGGGCGGGCTCTGCCGCCCCCACTGCCACCTAGTCTCTGAACCTTCTCCCACGGCACGACCCGTTTGGGAGCTTGGCTGCGGATTGCCCAATCTCAGACGTTTTTACCATACCCG(SEQ ID NO：3)。
由ITS序列分子生物学鉴定结果，可以确定该菌株为小球藻(Chlorella sp.)，分子生物学鉴定结果表明其与亲缘关系Micractinium pusillum(MK764926.1)的ITS序列相似性最高(相似性为98.58％，序列覆盖度为91％)，与Chlorella sorokiniana(KP645224.1)相似性为98.27％，序列覆盖度为90％，进化树结果表明该小球藻与Micractinium pusillum(MK764926.1)在进化树中分为一支(图2)，但形态上又有一定差异，我们将该藻株暂定为Chlorella sp.GB-116。
实施例2：小球藻GB-116(Chlorella sp.GB-116)的培养和藻粉的制备
1.种子培养
培养基：CaCl2·2H2O 0.2～0.4g/L、FeSO4·7H2O 5～25mg/L、MgSO4·7H2O0.05～0.2g/L、NaNO31.5～2.5g/L、K2HPO4 0.2～0.5g/L、酵母提取物0.5～1.5g/L、葡萄糖4～15g/L，琼脂1～3％，pH＝6.0～8.0。在本实施例中，采用的培养基的组成为：CaCl2·2H2O 0.3g/L、FeSO4·7H2O 15mg/L、MgSO4·7H2O 0.1g/L、NaNO3 2g/L、K2HPO4 0.3g/L、酵母提取物1g/L、葡萄糖8g/L、琼脂1.5％，pH＝6.0～8.0，加纯净水定容至1L。在固体培养基平板上划线得到单个藻落，在23～26℃、光照强度为3500～4000lx条件下静置培养3～5天。
2.种子的活化
从培养基平板上挑取单个藻落，接种到5ml异养培养基上，在23～26℃、光照强度为3500～4000lx的条件下静置培养3～5天。所述异养培养基的配方为：CaCl2·2H2O 0.2～0.4g/L、FeSO4·7H2O 5～25mg/L、MgSO4·7H2O 0.05～0.2g/L、NaNO31.5～2.5g/L、K2HPO40.2～0.5g/L、酵母提取物0.5～1.5g/L、葡萄糖4～15g/L，pH＝6.0～8.0。在本实施例中，异养培养基的组成为：CaCl2·2H2O0.3g/L、FeSO4·7H2O 15mg/L、MgSO4·7H2O 0.1g/L、NaNO32g/L、K2HPO40.3g/L、酵母提取物1g/L、葡萄糖8g/L；加纯净水定容到1L，pH＝6.0～8.0。
3.扩大培养
该小球藻经过100ml→5L→1000L的顺序进行扩大培养。扩大培养条件为23～25℃、光照强度3500～4000lx条件下，采用生物反应器在120～160rpm转速下培养。
4.小球藻藻粉的制备
藻体浓缩：采用中空纤维微滤膜过滤的方法浓缩小球藻培养液获得小球藻藻粉。所述中空纤维膜元件为PS材质，微滤膜组件规格为：
截留孔径为0.1～0.3μm，单根实际产水流量≥5000l/h、25℃，设计产水流量≥1200l/h、25℃。回收率为80～90％。
藻体干燥：浓缩的藻体在15～20℃条件下自然风干得到小球藻藻粉。
5.小球藻藻粉营养成分检测
藻粉总蛋白含量检测用凯氏定氮法(参见文献1)，总脂肪检测用索氏提取法(参见文献2)，17种水解氨基酸、β-胡萝卜素、虾青素含量检测方法用高效液相色谱法(参见文献3、4)，多糖、甾醇含量检测用紫外吸收法(参见文献5、6)，脂肪酸含量检测则选用气相色谱-质谱法(参见文献4)。
文献1：陈军,刘晶,杜泽鹏,等.索氏法提取植物种子粗脂肪的实验方法改进[J].轻工科技,2018,34(01):6-7.
文献2：Li Y,Sommerfeld M,Chen F,et al.Effect of photon flux densitieson regulation of carotenogenesis and cell viability of Haematococcuspluvialis(Chlorophyceae)[J].Journal of Applied Phycology,2010,22(3):253-263.
文献3：张玲.小球藻(Chlorella sorokiniana C74)的培养及活性物质的研究[D]；海南大学,2015.
文献4：Wang B,Zhang Z,Hu Q,et al.Cellular Capacities for High-LightAcclimation and Changing Lipid Profiles across Life Cycle Stages of the GreenAlga Haematococcus pluvialis[J].Plos One,2014,9(9):e106679.
文献5：柳小兰,张明,周美,等.西南红山茶茶籽中总甾醇的含量测定[J].食品科技,2017,42(01):260-263.
文献6：Luo W,Proschold T,Bock C,et al.Generic concept in Chlorella-related coccoid green algae(Chlorophyta,Trebouxiophyceae)[J].Plant Biology,2010,12(3):545-553.
结果表明，该藻粉的各营养成分含量如下述表1所示，其中总蛋白含量28.4％，多糖含量为1.29％，总脂肪含量1.1％，β-胡萝卜素含量127μg/g，虾青素含量0.850μg/g，总甾醇含量0.30％。该小球藻粉中含有多种人体必需氨基酸，具体的种类和含量如图3所示，其中蛋氨酸(甲硫氨酸)含量最高，达到36.205mg/g，其次是苏氨酸、丙氨酸、亮氨酸、赖氨酸、蛋氨酸和赖氨酸。该小球藻粉中含有15种脂肪酸种类分别为：C12:0(月桂酸)、C14:0(肉蔻酸)、C15:0(十五碳酸)、C16:0(棕榈酸)、C16:1(棕榈油酸)、C17:0(十七碳酸)、C17:1(十七碳一烯酸)、C18:0(硬脂酸)、C18:1N9C(油酸)、C18:1N9T(反式油酸)、C18:2N6T(反式亚油酸)、C18:3N3(α-亚麻酸)、C18:3N6(γ-亚麻酸)、C20:0(花生酸)、C24:0(二十四碳酸)，它们的含量如下述表2所示。其中C16:0(棕榈酸)含量最高，其次是C18:3N3(α-亚麻酸)、C18:1N9T(反式油酸)、C18:2N6T(反式亚油酸)，含量最低的是C12:0(月桂酸)。
表1：小球藻GB-116(Chlorella sp.GB-116)藻粉中各营养成分含量表
表2：小球藻GB-116(Chlorella sp.GB-116)藻粉中15种脂肪酸组成及含量
实施例3：小球藻GB-116(Chlorella sp.GB-116)藻粉作为饲料添加剂可提高鱼的免疫指标。
1.小球藻藻粉作为饲料添加剂制备水产颗粒饲料
水产基础饲料配方：鱼粉，20g；玉米粉，14g；豆粕，30g；花生麸，10g；米糠，2.5g；面粉，12.3g；虾壳粉，5g；磷脂油，0.5g；豆油，3g；磷酸二氢钙，2g；阿尔法淀粉，0.2g；多维多矿混料，0.5g。
对于免疫指标实验：以实施例2制备的小球藻藻粉为饲料添加剂根据配方制作三种饲料，小球藻粉藻添加量分别为0.5％(质量，下同)和2％(质量，下同)，G2A组未添加小球藻粉(对照组)，G2B组添加0.5％小球藻粉(试验组)，G2D组添加2％小球藻粉(试验组)。准备好各种材料，用地臣双齿劲锤粉碎机将豆粕和花生麸粉碎，各组分原料过40目筛备用，之后用电子天平准确称量各原料，倒入盆中均匀混合，再加入磷脂油和豆油充分混匀，最后加入适量水混合均匀后，用地臣SKJ系列平模颗粒机制成粒直径2mm，长度<4mm的颗粒饲料，与室温下用电风扇风干，装入密封袋中，放于-20℃储存。
2.实验用罗非鱼
试验罗非鱼购自钦州罗非鱼养殖场，喂养前对鱼和养殖系统进行消毒处理，先用商品饲料喂养一段时间后，再挑选体格健壮、健康活泼、规格一致的罗非鱼170尾，用对照组饲料驯养7天。随机分为3组(1个对照和2个试验组)，初始体重(6.64±0.88)g，每组3个重复，随机分到9个规格相同的鱼桶里(每个鱼桶18条鱼)。养殖实验在北部湾大学海洋学院循环水养殖实验室进行。
3.罗非鱼的饲喂
室内循环水喂养罗非鱼，采用三个水循环养殖系统(各含一个水净化系统)，每个养殖系统有3个桶(80cm×80cm)，分别养殖对照组，两个试验组。每天投喂两次，时间分别为8:00和17:00，日投喂量为体重的2～3％(质量)，每天记录水温、死亡条数及投喂饲料量，室温保持24℃，水温22-24℃，保持鱼桶内连续不断充氧，保持溶氧量﹥5mg/L，pH＝7.0～7.5，氨氮<0.04mg/L，每2天换1次水，换水量为1/3，养殖周期为4周。
4.实验用鱼样品的采集
基础饲料驯养结束后，养殖过程中第15天每组随机选取6尾鱼(每个桶挑选2尾)，解剖取鱼肝胰脏样本放于无菌1.5ml尖底离心管中置于液氮冷冻，之后将所有样品放于超低温冰箱短暂保存。之后用干冰将样品送至天津诺禾致源生物信息科技有限公司进行转录组测序。提取RNA样品时每组6条鱼，取部分肝胰腺样品两两混合提取，样品编号如表3所示，G2A组未添加小球藻粉(对照组)，G2B组添加0.5％小球藻粉，G2D组添加2％小球藻粉。
表3：各组样品编号记录表
5.转录组测序分析
肝胰脏转录组测序分析流程图如图4所示，差异基因筛选采用软件DESeq2，显著性差异基因筛选阈值为padj<0.05和|log2FoldChange|>0.0，clusterProfiler软件分析GO功能富集和KEGG通路富集分析以padj小于0.05作为显著性富集的阈值，padj为多重假设检验校正后的p值。
结果表明，对添加了2％小球藻藻粉的实验组(G2D组)和未添加藻粉的对照组(G2A组)的差异基因进行GO功能富集分析发现(padj小于0.05作为显著性富集的阈值，padj为多重假设检验校正后的p值)，与对照相比，添加了2％藻粉的饲料的鱼，其肝脏中有11个与免疫相关的基因显著性富集：5个与细胞粘附有关(padj在0.001到0.006之间)，2个与免疫相关(padj<0.5),富集的基因其功能主要集中在血红素结合、铁离子结合、钙离子结合、丝氨酸型内肽酶抑制剂等功能性蛋白。
同时在各分组在初重无显著差异的情况下，饲喂15天后，不添加小球藻藻粉的对照组(W1A)体重平均值为10.16克，添加了0.5％的小球藻藻粉的组(W1B)，鱼体重为15.72克，添加了2％的小球藻藻粉的组(W1C)，鱼体重为15.88克；经过One-way ANOVA方法统计分析发现，W1B和W1A组的平均体重存在显著差异(p<0.05)，W1C和W1A组的平均体重也存在显著差异(p<0.05)，如图5所示。可见，小球藻藻粉添加到饲料中能够提高幼鱼的生长性能。
实施例4：小球藻GB-116(Chlorella sp.GB-116)藻粉作为饲料添加剂可改善鱼肠道菌群
1.小球藻藻粉作为饲料添加剂制备水产颗粒饲料
水产基础饲料配方：同实施例3。
以实施例2制备的小球藻藻粉为饲料添加剂根据配方制作三种饲料，小球藻粉藻添加量分别为0.5％(质量，下同)和2％(质量，下同)，C2A组未添加小球藻粉(对照组)，C2B组添加0.5％小球藻粉(试验组)，C2D组添加2％小球藻粉(试验组)。准备好各种材料，用地臣双齿劲锤粉碎机将豆粕和花生麸粉碎，各组分原料过40目筛备用，之后用电子天平准确称量各原料，倒入盆中均匀混合，再加入磷脂油和豆油充分混匀，最后加入适量水混合均匀后，用地臣SKJ系列平模颗粒机制成粒直径2mm，长度<4mm的颗粒饲料，与室温下用电风扇风干，装入密封袋中，放于-20℃储存。
2.实验用罗非鱼
试验罗非鱼购自钦州罗非鱼养殖场，喂养前对鱼和养殖系统进行消毒处理，先用商品饲料喂养一段时间后，再挑选体格健壮、健康活泼、规格一致的罗非鱼170尾，用对照组饲料驯养7天。然后将鱼随机分为3组(1个对照和2个试验组)，初始体重(6.64±0.88)g，每组3个重复，随机分到9个规格相同的鱼桶里(每个鱼桶18条鱼)。养殖实验在北部湾大学海洋学院循环水养殖实验室进行。
3.罗非鱼的的饲喂
4.室内循环水喂养罗非鱼，采用三个水循环养殖系统(各含一个水净化系统)，每个养殖系统有3个桶(80cm×80cm)，分别养殖对照组，两个试验组。每天投喂两次，时间分别为8:00和17:00，日投喂量为体重的2～3％(质量)，每天记录水温、死亡条数及投喂饲料量，室温保持24℃，水温22～24℃，保持鱼桶内连续不断充氧，保持溶氧量﹥5mg/L，pH＝7.0～7.5，氨氮<0.04mg/L，每2天换1次水，换水量为1/3，养殖周期为4周。
5.实验用鱼样品的采集
养殖试验开始前，随机选5尾罗非鱼取初始肠道样品，养殖过程中，养殖中期和后期各测一次鱼体重，第15天每组随机选取6尾鱼(每个桶挑选2尾)解剖取鱼肠道内容物样本，于液氮冷冻后放置超低温冰箱短暂保存。之后用干冰将样品送至天津诺禾致源生物信息科技有限公司进行16s rRNA高通量测序。初始样品、对照组以及试验组肠道最终的检测样品编号如表4所示。
表4：各组样品编号记录表
6.罗非鱼肠道菌群16s rRNA分析
肠道样品16s rRNA测序分析流程如图6所示，进行样本检测、DNA提取与检测、PCR扩增及产物纯化、建库、测序等一系列操作，在基于Illumina Nova测序平台测序，将测序得到的原始数据经拼接、过滤处理后得到有效数据。Alpha多样性指数组间差异分析进行wilcox秩和检验显著性，以P小于0.05作为显著性富集的阈值；MetaStat分析以P-value小于0.05作为显著性富集的阈值，P-value值为多重检验校正的P值；LEfSe分析以LDA score﹥4筛选显著性差异物种的阈值。
结果表明，采用LEfSe分析不同饲料组间的物种丰度数据，检测不同饲料组间的差异物种，本实验以LDAscore﹥4筛选显著性差异物种，差异物种的LDA值分布柱状图如图7所示，通过对对照组(C2A组)和添加有2％小球藻藻粉(C2D组)差异物种的LEfSe分析，添加了2％藻粉的饲料可以明显富集肠道益生菌，增加肠球菌(Enterococcus)﹑乳酸菌(Lactobacillus)﹑粪球菌(Coprococcus)﹑蛭弧菌(Bdellovibrio)﹑双歧杆菌属(Bifidobacterium)的相对丰度。
序列表
<110> 福建师范大学
北部湾大学
<120> 小球藻的应用
<130> 2021
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 19
<212> DNA
<213> 人工序列()
<400> 1
tccgtaggtg aacctgcgg 19
<210> 2
<211> 20
<212> DNA
<213> 人工序列()
<400> 2
tcctccgctt attgatatgc 20
<210> 3
<211> 906
<212> DNA
<213> Chlorella sp. GB-116
<400> 3
cgaagtgaga gcgttcgggg ggccgaaacc accaccgaat tcctgctagg ccgccagcaa 60
agtcccctcg ggccacggca ggctgcgtag cgggtgctac ctaccaagcc attgccctgc 120
aaacggggtc catgctcaag cctctacact tcagccgacc cggacgcagc ctgtgacagg 180
caacggagcc gggaaggcca ggtccgcact ccctcaggga gaggggggcg agggtgtaag 240
ccgacgctga ggcagacatg ccctcggccg aagcctcggg cgcaatttgc gttcaaagat 300
tcgatggttc acggaattct gcaattcaca ctacgtatcg catttcgctg cgttcttcat 360
cgatacggga gccaagatat ccgttgttga gagttgtctt tggtttgtgc ggagcggaca 420
ggccgccacc gctgtttgct tcagagtggt tttgtgtttg gggtggattg agttggatac 480
aaattaccag cgtcaccagc cggcgagggc aaagcccaac gccggagaga cgcccgccgg 540
acctgaaact gcggctcgta agccccagcc aggggaccga cgcccggcgg gagccgcctt 600
agaaaaggca ggcccaacca cacggggggg acaaatggtt accagggtgg attcgatcga 660
ttcaatgatc cttccgcagg ttcacctcgg gtactcccac caacctggtc gacaggctgg 720
ctccctgggc tccgggctgg ttaggccgtt cctcccaggg caccctctcg ggtgggccgg 780
actctatctt aagggcgggc tctgccgccc ccactgccac ctagtctctg aaccttctcc 840
cacggcacga cccgtttggg agcttggctg cggattgccc aatctcagac gtttttacca 900
tacccg 906
Claims (3)
1.小球藻或小球藻藻粉在制备饲料添加剂中的应用，其中，
所述小球藻GB-116(Chlorella sp. GB-116)，其保藏编号为CCTCC No：M 2020373；
所述小球藻藻粉是将小球藻GB-116(Chlorella sp. GB-116)置于异养培养基中，在23~26℃、光照强度为3500~4000 lx的条件下进行培养；所得培养物经截留孔径为0.1~0.3μm的微滤膜过滤，收集截留物，收集截留物，干燥而得。
2.根据权利要求1所述的应用，其特征是，所述异养培养基为改良的克氏培养基，其组成为：CaCl2•2H2O 0.2~0.4 g/L、FeSO4•7H2O 5~25 mg/L、MgSO4•7H2O 0.05~0.2 g /L、NaNO31.5~2.5 g/L、K2HPO4 0.2~0.5 g/L、酵母提取物0.5~1.5 g/L、葡萄糖4~15 g/L，pH=6.0~8.0。
3.一种组合物，其包含小球藻的细胞和/或其提取物和/或其培养物，所述的组合物为饲料添加剂，所述的小球藻为小球藻GB-116(Chlorella sp. GB-116)，其保藏编号为CCTCCNo：M 2020373。
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110890806.2A CN113678942B (zh) | 2021-08-04 | 2021-08-04 | 小球藻的应用 |
| CN202310628284.8A CN116622513A (zh) | 2021-08-04 | 2021-08-04 | 一种小球藻、小球藻藻粉及其制备方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110890806.2A CN113678942B (zh) | 2021-08-04 | 2021-08-04 | 小球藻的应用 |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310628284.8A Division CN116622513A (zh) | 2021-08-04 | 2021-08-04 | 一种小球藻、小球藻藻粉及其制备方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113678942A CN113678942A (zh) | 2021-11-23 |
| CN113678942B true CN113678942B (zh) | 2023-06-06 |
Family
ID=78578780
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310628284.8A Pending CN116622513A (zh) | 2021-08-04 | 2021-08-04 | 一种小球藻、小球藻藻粉及其制备方法 |
| CN202110890806.2A Active CN113678942B (zh) | 2021-08-04 | 2021-08-04 | 小球藻的应用 |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310628284.8A Pending CN116622513A (zh) | 2021-08-04 | 2021-08-04 | 一种小球藻、小球藻藻粉及其制备方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN116622513A (zh) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114196544A (zh) * | 2021-08-04 | 2022-03-18 | 北部湾大学 | 一种小球藻、小球藻藻粉及其制备方法 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110604234A (zh) * | 2019-05-22 | 2019-12-24 | 中国科学院水生生物研究所 | 索罗金小球藻在改善鱼类健康状况的功能性饲料中的应用 |
| CN111925943B (zh) * | 2020-08-28 | 2023-01-10 | 河北工业大学 | 天津拟甲小球藻及其培养方法与应用 |
-
2021
- 2021-08-04 CN CN202310628284.8A patent/CN116622513A/zh active Pending
- 2021-08-04 CN CN202110890806.2A patent/CN113678942B/zh active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN113678942A (zh) | 2021-11-23 |
| CN116622513A (zh) | 2023-08-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Abomohra et al. | Pilot cultivation of the chlorophyte microalga Scenedesmus obliquus as a promising feedstock for biofuel | |
| CN107287252B (zh) | 一种ω-7脂肪酸合成物及培养黄丝藻生产该合成物的方法与应用 | |
| DK2895628T3 (da) | Olie beriget på arachidonsyre fra mikroorganismer (encellet mortierella alpina-svamp) og fremgangsmåde til fremstilling heraf | |
| CN112226373B (zh) | 一株产蛋白的菌株及其应用 | |
| Fan et al. | Production of high-value products by marine microalgae thraustochytrids | |
| WO2012175027A1 (en) | Methods of mutagenesis of schizochytrium sp and variant strains produced thereof | |
| JP2020501547A (ja) | バイオマス材料からの核酸及びそのフラグメントの除去 | |
| CN101020895B (zh) | 一种植物乳杆菌发酵制备富含共轭亚油酸青贮饲料的方法 | |
| WO2012047120A1 (en) | Heterotrophic microbial production of xanthophyll pigments | |
| Gonzalez et al. | Bioprospection of green microalgae native to Paraná, Brazil using a multi-criteria analysis: Potential for the production of lipids, proteins, and carotenoids | |
| Lu et al. | Biomass and nutrient productivities of Tetraselmis chuii under mixotrophic culture conditions with various C: N ratios | |
| CN113678942B (zh) | 小球藻的应用 | |
| CN115418320B (zh) | 一株高产蛋白质的蛋白核小球藻及其培养方法和应用 | |
| CA2904038C (en) | Production of omega-3 fatty acids from pythium species | |
| RU2573944C1 (ru) | Штамм микроводоросли haematococcus pluvialis - продуцент натурального астаксантина | |
| CN110089623B (zh) | 栅藻及其作为饵料/饵料添加剂的应用 | |
| CN114196544A (zh) | 一种小球藻、小球藻藻粉及其制备方法 | |
| JP2022541726A (ja) | 家畜及び魚の飼料タンパク質添加物 | |
| CN116762898B (zh) | 基于CpG寡核苷酸串联分子的对虾免疫增强剂及其应用 | |
| CN115161201B (zh) | 一种栅列藻藻株及其培养方法和用途 | |
| KR102242665B1 (ko) | 클로렐라 불가리스 hsp70 유래의 열 유도성 프로모터 및 이의 용도 | |
| CN116640687B (zh) | 罗姆布茨菌my01及其应用 | |
| KR20130048942A (ko) | 성장률이 우수한 나노클로롭시스 속 신균주 및 이의 용도 | |
| WO2024147286A1 (ja) | 微細藻類増殖促進用微生物、微細藻類増殖促進剤、微細藻類の培養方法、微生物のスクリーニング方法 | |
| Chakraborty et al. | MICROALGAE: UTILITIES AND PRODUCTION FOR COMMERCIALIZATION |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |