WO2007046685A1 - Starter kit for the production of pure and high quality microalgae - Google Patents
Starter kit for the production of pure and high quality microalgae Download PDFInfo
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- WO2007046685A1 WO2007046685A1 PCT/MY2006/000019 MY2006000019W WO2007046685A1 WO 2007046685 A1 WO2007046685 A1 WO 2007046685A1 MY 2006000019 W MY2006000019 W MY 2006000019W WO 2007046685 A1 WO2007046685 A1 WO 2007046685A1
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- Prior art keywords
- spp
- liquid concentrate
- microalgae
- nutrient rich
- powder
- Prior art date
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- 238000004519 manufacturing process Methods 0.000 title claims description 10
- 239000007858 starting material Substances 0.000 title description 3
- 239000001963 growth medium Substances 0.000 claims abstract description 26
- 235000015097 nutrients Nutrition 0.000 claims abstract description 24
- 239000000843 powder Substances 0.000 claims abstract description 22
- 239000010802 sludge Substances 0.000 claims abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 238000001914 filtration Methods 0.000 claims abstract description 8
- 230000001954 sterilising effect Effects 0.000 claims abstract description 7
- 239000011148 porous material Substances 0.000 claims abstract description 4
- 235000014666 liquid concentrate Nutrition 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 16
- 238000004108 freeze drying Methods 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 4
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- 230000006835 compression Effects 0.000 claims description 3
- 235000016709 nutrition Nutrition 0.000 claims description 3
- 241000611184 Amphora Species 0.000 claims description 2
- 235000016425 Arthrospira platensis Nutrition 0.000 claims description 2
- 240000002900 Arthrospira platensis Species 0.000 claims description 2
- 241000227752 Chaetoceros Species 0.000 claims description 2
- 241000195649 Chlorella <Chlorellales> Species 0.000 claims description 2
- 241000224474 Nannochloropsis Species 0.000 claims description 2
- 241000192497 Oscillatoria Species 0.000 claims description 2
- 241000192511 Pseudanabaena Species 0.000 claims description 2
- 241000206733 Skeletonema Species 0.000 claims description 2
- 241000196321 Tetraselmis Species 0.000 claims description 2
- 239000003365 glass fiber Substances 0.000 claims description 2
- 229940082787 spirulina Drugs 0.000 claims description 2
- 238000005374 membrane filtration Methods 0.000 claims 2
- 241001501885 Isochrysis Species 0.000 claims 1
- 241000502321 Navicula Species 0.000 claims 1
- 238000002156 mixing Methods 0.000 claims 1
- 239000013049 sediment Substances 0.000 abstract description 10
- 239000002609 medium Substances 0.000 abstract description 9
- 239000007788 liquid Substances 0.000 abstract description 7
- 238000012258 culturing Methods 0.000 abstract description 6
- 238000007865 diluting Methods 0.000 abstract description 3
- 239000012141 concentrate Substances 0.000 abstract description 2
- 238000004806 packaging method and process Methods 0.000 abstract 1
- 238000009360 aquaculture Methods 0.000 description 10
- 244000144974 aquaculture Species 0.000 description 10
- 239000000126 substance Substances 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 241000195493 Cryptophyta Species 0.000 description 6
- MBMBGCFOFBJSGT-KUBAVDMBSA-N all-cis-docosa-4,7,10,13,16,19-hexaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 description 6
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 5
- 238000011109 contamination Methods 0.000 description 4
- 239000011785 micronutrient Substances 0.000 description 4
- 235000021049 nutrient content Nutrition 0.000 description 4
- 241000091752 Chaetoceros calcitrans Species 0.000 description 3
- 229940090949 docosahexaenoic acid Drugs 0.000 description 3
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 3
- 239000003337 fertilizer Substances 0.000 description 3
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- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 241000251468 Actinopterygii Species 0.000 description 2
- 241000238557 Decapoda Species 0.000 description 2
- 241000159660 Nannochloropsis oculata Species 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000005416 organic matter Substances 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
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- 229920001817 Agar Polymers 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000195628 Chlorophyta Species 0.000 description 1
- 241001607798 Cymbella Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000192520 Oscillatoria sp. Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241000192500 Spirulina sp. Species 0.000 description 1
- 241000450725 Tetraselmis tetrathele Species 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- YYRMJZQKEFZXMX-UHFFFAOYSA-N calcium;phosphoric acid Chemical compound [Ca+2].OP(O)(O)=O.OP(O)(O)=O YYRMJZQKEFZXMX-UHFFFAOYSA-N 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- -1 carbons Chemical class 0.000 description 1
- 235000008504 concentrate Nutrition 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000021112 essential micronutrients Nutrition 0.000 description 1
- 235000021321 essential mineral Nutrition 0.000 description 1
- 238000012851 eutrophication Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
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- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
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- 239000000377 silicon dioxide Substances 0.000 description 1
- LIVNPJMFVYWSIS-UHFFFAOYSA-N silicon monoxide Chemical class [Si-]#[O+] LIVNPJMFVYWSIS-UHFFFAOYSA-N 0.000 description 1
- 229910052814 silicon oxide Inorganic materials 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000002426 superphosphate Substances 0.000 description 1
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- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
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
Definitions
- the present invention relates to a method used for preparation of culture medium for microalgae from marine aquaculture pond sludge.
- Said culture medium in turn can be used as a starter kit to initialize the growth of microalgae culture with pure microalgae isolates to produce high quality microalgae.
- microalgae have high contents of important polyunsaturated fatty acids such as arachidonic acid (20:4n-6), EPA (20:5n- 5), DPA (22:5n-3) and DHA (22:6n-3).
- arachidonic acid (20:4n-6), EPA (20:5n- 5), DPA (22:5n-3) and DHA (22:6n-3).
- microalgae are excellence sources of important nutrients in human and animal health.
- it also tiihances the fish colour within the water habitats as well as being the live food for fish and shrimp larvae production.
- pure and high quality microalgae are commonly used for feed additives, pharmaceutical, neutraceutical and cosmetic products.
- fertilizer for the production of microalgae results in contamination such as heavy metals, antibiotics and other harmful organisms.
- the quality of algae produced is low and variable, depending on the source of organic fertilizer.
- chemical fertilizers such as urea or triple super phosphate would not provide all the essential ingredients such as trace minerals found in interstitial water.
- a complete culture medium for microalgae widely used in laboratories has to be made up of 15 different chemicals. Thus it is essential to develop a cheap and easy way to start a pure algae culture.
- Bottom sediment from aquaculture ponds generally receives large amount of high quality organic matter in the form of feed, fertilizers and excretory products resulting in enriched sludge. Although those bottom sediment sludge is rich in phosphates, nitrogen, carbons, silicon oxides and other micronutrients, untreated aquaculture sludge is actually a source of pollution to the aquatic environment. Discharge of this enriched sludge can cause serious eutrophication problems in public waters. This can undermine the sustainability of all aquatic-based industries including aquaculture.
- treatment or recycling of said sludge can be done by extracting the interstitial water containing these nutrients. Discharge of said treated
- the present invention provides a method for production of microalgae culture medium comprising the steps of obtaining pond sludge from aquaculture pond sediment followed by extracting the interstitial water through centrifiigation or compression, filtering and sterilizing the extract and finally freeze-drying the concentrate extract to become powder.
- the produced culture medium can be used to start pure cultures with minimal contamination.
- the main objective of said invention is to provide a cheap and easy method to produce pure and high quality microalgae. This is because obtaining and maintaining pure microalgae isolates are difficult, it requires expertise and special equipment. This culture medium preparation steps are relatively easy and its production cost is much cheaper than standard medium. The produced culture medium is made to powder form for easy handling and storage.
- the culture medium prepared according to the disclosed method can initialize pure microalgae culture by adding the provided pure microalgae isolates into the diluted said culture medium in desired concentration. Mass culture of pure microalgae is made easy with minimal contamination. Microalgae grown in the said culture medium contains higher levels of nutritional value comparing to those grown in standard medium.
- Figure 1 a schematic flow-chart of the disclosed method.
- Present invention provides a method for preparing culture medium from aquaculture pond bottom sludge for culturing pure microalgae species.
- the nutrient-rich interstitial water (IW) extracted from aquaculture pond bottom sediments is collected and undergone extraction followed by filtration and sterilization to obtain a nutrient concentrated extract.
- the nutrient concentrated extract is made to powder form by freeze-drying. Customized pure microalgae isolates is supplemented to said concentrated extract or powder to provide a microalgae growth initializing kit for microalgae culturing purposes.
- the steps of disclosed invention are illustrated as in Figure 1.
- the enriched aquaculture pond bottom sediment or referred as pond sludge hereafter, is collected either manually or with the aid of a suction pump.
- a kilogram of pond sludge can produce 500-1000 ml of concentrated liquid extract depending on the texture of the sediment.
- the amount of pond sludge to be collected is depending on the
- the collected pond sludge is subjected to extraction to obtain the pore water or interstitial water (IW), referred as liquid concentrate hereafter.
- the sludge is undergone centrifugation (using a centrifuge) at the speed of 6000 to 8000 rpm for 20 to 40 minutes. Centrifugation is undergone only once to extract the pore water or IW from each loading of pond sludge in the centrifuge container.
- the sludge can be undergone compression (using a compressor) to sediment down the sludge leaving the said liquid concentrate which is rich in micronutrients. During the process, no other additional chemical is added.
- said liquid concentrate is transferred to a clean container.
- Filtration is carried out through glass fiber filter followed by 0.45 and 0.22 ⁇ m membrane filters.
- Said filtering step is to filter out the bacteria and other living microorganisms. This step is best done in a sterile condition.
- Said filtered liquid concentrate is then subjected to chemical analysis to determine its nutrient content.
- UV light far-UV range of 254 nm
- Sterilization by this method but not autoclaving is to ensure all the enrich media are maintained and not destroyed by heat and pressure due to autoclave.
- the sterilized liquid extract is ready to use upon dilution to desired concentration depending on the nutrient content
- Said sterile liquid concentrate is transferred aseptically to ice cube maker to form ice cubes.
- Said liquid concentrated in ice cube form is then transferred in a sterile plastic sheet and placed into the freeze drier container.
- Said ice cube is freeze dried within 3 days in freeze drier at the temperature of -40 to -55 0 C and vacuum pressure of 130- 150 ⁇ l0 3 mbar.
- the nutrient rich powder obtained is ready to use after freeze drying.
- Said nutrient rich powder is stored in air tight containers at 20 to 30 0 C. Depending on the determined nutrient content, 1 g of said nutrient rich powder can produce 5 to 10 liter of liquid culture media.
- Said liquid culture medium is stored in air tight bottles and store at 2-6 °C.
- the nutrient rich liquid concentrate or powder can be further processed in the form of a kit, a microalgae growth initializing kit.
- the kit comprises the nutrient rich liquid concentrate or powder and several types of customized microalgae isolates. Said customized microalgae isolates can be mixed with the nutrient rich liquid concentrate or powder and users can simply culture the customized microalgae by diluting said mixture according to the instruction given.
- the abovementioned customized microalgae are Chaetoceros spp., Skeletonema spp., Isochtysis spp., Amphora spp., Cymbella spp, Naviciila spp., Chlorella spp., Nannochloropsis spp., Tetraselmis spp., Oscillatoria spp. Spirulina spp., and Pseudanabaena spp.
- said customized microalgae isolates can be provided as additional supplement without the nutrient rich liquid concentrate or powder or vice versa.
- the disclosed kit is ready to be used by anyone with or without expertise in culturing microalgae as the culturing step
- the culture medium produced from the nutrient rich liquid concentrate or powder is rich in phosphorus, nitrogen, silica, essential minerals and micronutrients that meet the requirement for healthy production of high quality algal species.
- Said culture medium above can be used for mass culture of a pure species without contamination.
- pure algal cultures will develop profusely reaching a plateau within 7 to 10 days.
- marine algae Choetoceros calcitrans (a diatom), Oscillatoria sp.
- the graph below shows the culture of a diatom, Chaetoceros calcitrans and green algae, Nannochloropsis oculata in standard culture medium (Conway) and diluted interstitial water (DIW).
- diluted algal culture medium was the best medium for culturing pure stocks of microalgae.
- Algae cultured in the said culture medium showed significantly higher nutritional quality than those grown in standard (Conway) medium.
- the cultured algae contain higher concentration of essential amino acids and polyunsaturated fatty acids (especially omega 3 and 6), compared to those grown in standard Conway medium which contains about 15 different chemicals.
- the graph below shows the fatty acid (g/lOOg lipids) profile of a diatom, Chaetoceros calcitrans in standard culture medium (Conway) and diluted interstitial water (DIW).
- microalgae isolates are stored in dark bottles at 2-6 0 C and can last for minimum 6 months.
- Algal culture can be easily initiated by diluting the liquid extract or powder in transparent containers such as plastic bags or bottles followed by inoculating algal cells provided.
- Said microalgae culture medium can be used as solid medium 'for growing microalgal stocks by adding agar into it.
- the filtered liquid extract is subjected to chemical analysis to determine its nutrient content.
- Table below shows the chemical and physical parameters of said filtered extract from shrimp ponds and Conway medium. Values are mean ⁇ standard deviation.
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Abstract
A novel use of nutrient rich pore water or interstitial water (IW) extracted from aquaculrure pond bottom sediments is an alternative medium for culturing different pure microalgal species. The medium is prepared by extracting, filtering, sterilizing the interstitial water collected from enriched sludge sediment and finally concentrate the interstitial water to nutrient rich liquid extract or powder forms to be used as culture medium by diluting or dissolving it, respectively in water. A microalgae growth initializing kit is produced by packaging microalgae isolates with the nutrient rich liquid extract and/or powder.
Description
Starter Kit for the Production of Pure and High Quality Microalgae
Field of Invention
The present invention relates to a method used for preparation of culture medium for microalgae from marine aquaculture pond sludge. Said culture medium in turn can be used as a starter kit to initialize the growth of microalgae culture with pure microalgae isolates to produce high quality microalgae.
Background of the Invention
Pure algae are actually in great demand, especially in the field of aquaculture industry, research institutions and schools. This is because pure microalgae have high contents of important polyunsaturated fatty acids such as arachidonic acid (20:4n-6), EPA (20:5n- 5), DPA (22:5n-3) and DHA (22:6n-3). Hence, microalgae are excellence sources of important nutrients in human and animal health. Furthermore, it also tiihances the fish colour within the water habitats as well as being the live food for fish and shrimp larvae production. Besides, pure and high quality microalgae are commonly used for feed additives, pharmaceutical, neutraceutical and cosmetic products.
Due to the fact that obtaining and maintaining pure microalgae isolates are difficult because it requires expertise and special equipment. For example, the use of organic
fertilizer for the production of microalgae results in contamination such as heavy
metals, antibiotics and other harmful organisms. In addition, the quality of algae produced is low and variable, depending on the source of organic fertilizer. The use of chemical fertilizers such as urea or triple super phosphate would not provide all the essential ingredients such as trace minerals found in interstitial water. In fact, a complete culture medium for microalgae widely used in laboratories has to be made up of 15 different chemicals. Thus it is essential to develop a cheap and easy way to start a pure algae culture.
Bottom sediment from aquaculture ponds generally receives large amount of high quality organic matter in the form of feed, fertilizers and excretory products resulting in enriched sludge. Although those bottom sediment sludge is rich in phosphates, nitrogen, carbons, silicon oxides and other micronutrients, untreated aquaculture sludge is actually a source of pollution to the aquatic environment. Discharge of this enriched sludge can cause serious eutrophication problems in public waters. This can undermine the sustainability of all aquatic-based industries including aquaculture.
On the other hand, organic matter from aquaculture practices enriched the bottom sediment. The availability of growth nutrients in the interstitial water extracted from the sludge, such as nitrogen, phosphorus and other micronutrients can be used to produce
high quality microalgae. In fact, treatment or recycling of said sludge can be done by extracting the interstitial water containing these nutrients. Discharge of said treated
sludge would minimize pollution in rivers and coastal areas.
Summary of Invention
The present invention provides a method for production of microalgae culture medium comprising the steps of obtaining pond sludge from aquaculture pond sediment followed by extracting the interstitial water through centrifiigation or compression, filtering and sterilizing the extract and finally freeze-drying the concentrate extract to become powder. The produced culture medium can be used to start pure cultures with minimal contamination.
The main objective of said invention is to provide a cheap and easy method to produce pure and high quality microalgae. This is because obtaining and maintaining pure microalgae isolates are difficult, it requires expertise and special equipment. This culture medium preparation steps are relatively easy and its production cost is much cheaper than standard medium. The produced culture medium is made to powder form for easy handling and storage.
Apart from that, the culture medium prepared according to the disclosed method can initialize pure microalgae culture by adding the provided pure microalgae isolates into the diluted said culture medium in desired concentration. Mass culture of pure microalgae is made easy with minimal contamination. Microalgae grown in the said culture medium contains higher levels of nutritional value comparing to those grown in standard medium.
Brief Description of Figures
Figure 1 a schematic flow-chart of the disclosed method.
Detailed Description of Invention
Present invention provides a method for preparing culture medium from aquaculture pond bottom sludge for culturing pure microalgae species. The nutrient-rich interstitial water (IW) extracted from aquaculture pond bottom sediments is collected and undergone extraction followed by filtration and sterilization to obtain a nutrient concentrated extract. The nutrient concentrated extract is made to powder form by freeze-drying. Customized pure microalgae isolates is supplemented to said concentrated extract or powder to provide a microalgae growth initializing kit for microalgae culturing purposes. The steps of disclosed invention are illustrated as in Figure 1.
First, the enriched aquaculture pond bottom sediment, or referred as pond sludge hereafter, is collected either manually or with the aid of a suction pump. A kilogram of pond sludge can produce 500-1000 ml of concentrated liquid extract depending on the texture of the sediment. The amount of pond sludge to be collected is depending on the
amount of extract required. ■
The collected pond sludge is subjected to extraction to obtain the pore water or interstitial water (IW), referred as liquid concentrate hereafter. The sludge is undergone centrifugation (using a centrifuge) at the speed of 6000 to 8000 rpm for 20 to 40 minutes. Centrifugation is undergone only once to extract the pore water or IW from each loading of pond sludge in the centrifuge container. Alternatively, the sludge can be undergone compression (using a compressor) to sediment down the sludge leaving the said liquid concentrate which is rich in micronutrients. During the process, no other additional chemical is added.
After the extraction process, said liquid concentrate is transferred to a clean container. Filtration is carried out through glass fiber filter followed by 0.45 and 0.22 μm membrane filters. Said filtering step is to filter out the bacteria and other living microorganisms. This step is best done in a sterile condition. Said filtered liquid concentrate is then subjected to chemical analysis to determine its nutrient content.
Next, said analyzed' liquid concentrate is subjected to sterilization through ultraviolet (UV) light (far-UV range of 254 nm) exposure for 1-2 hours. UV light eradicates all those smaller microorganisms which are not being filtered out previously. Sterilization by this method but not autoclaving is to ensure all the enrich media are maintained and not destroyed by heat and pressure due to autoclave. The sterilized liquid extract is ready to use upon dilution to desired concentration depending on the nutrient content
determined previously.
Said sterile liquid concentrate is transferred aseptically to ice cube maker to form ice cubes. Said liquid concentrated in ice cube form is then transferred in a sterile plastic sheet and placed into the freeze drier container. Said ice cube is freeze dried within 3 days in freeze drier at the temperature of -40 to -55 0C and vacuum pressure of 130- 150χl03 mbar. The nutrient rich powder obtained is ready to use after freeze drying. Said nutrient rich powder is stored in air tight containers at 20 to 30 0C. Depending on the determined nutrient content, 1 g of said nutrient rich powder can produce 5 to 10 liter of liquid culture media. Said liquid culture medium is stored in air tight bottles and store at 2-6 °C.
Moreover, the nutrient rich liquid concentrate or powder can be further processed in the form of a kit, a microalgae growth initializing kit. The kit comprises the nutrient rich liquid concentrate or powder and several types of customized microalgae isolates. Said customized microalgae isolates can be mixed with the nutrient rich liquid concentrate or powder and users can simply culture the customized microalgae by diluting said mixture according to the instruction given. The abovementioned customized microalgae are Chaetoceros spp., Skeletonema spp., Isochtysis spp., Amphora spp., Cymbella spp, Naviciila spp., Chlorella spp., Nannochloropsis spp., Tetraselmis spp., Oscillatoria spp. Spirulina spp., and Pseudanabaena spp. In another preferred embodiment, said customized microalgae isolates can be provided as additional supplement without the nutrient rich liquid concentrate or powder or vice versa. The disclosed kit is ready to be used by anyone with or without expertise in culturing microalgae as the culturing step
can be easily followed according to the instruction enclosed with the kit.
Example 1
The culture medium produced from the nutrient rich liquid concentrate or powder is rich in phosphorus, nitrogen, silica, essential minerals and micronutrients that meet the requirement for healthy production of high quality algal species. Said culture medium above can be used for mass culture of a pure species without contamination. On the exposure of the plastic bags to sunlight or other light sources, pure algal cultures will develop profusely reaching a plateau within 7 to 10 days. Experiments showed that marine algae (Chaetoceros calcitrans (a diatom), Oscillatoria sp. (a blue-green alga) and Nannochloropsis oculata (a green alga), Tetraselmis tetrathele, and Spirulina sp.) cultured in said culture medium produced significantly higher density (p<0.05) and culture rate than the standard culture medium (Conway). The graph below shows the culture of a diatom, Chaetoceros calcitrans and green algae, Nannochloropsis oculata in standard culture medium (Conway) and diluted interstitial water (DIW).
Days of culture
Example 2
Further bioassays demonstrated that diluted algal culture medium was the best medium for culturing pure stocks of microalgae. Algae cultured in the said culture medium showed significantly higher nutritional quality than those grown in standard (Conway) medium. For example, the cultured algae contain higher concentration of essential amino acids and polyunsaturated fatty acids (especially omega 3 and 6), compared to those grown in standard Conway medium which contains about 15 different chemicals. The graph below shows the fatty acid (g/lOOg lipids) profile of a diatom, Chaetoceros calcitrans in standard culture medium (Conway) and diluted interstitial water (DIW). (EPA: eicosapεntaenoic acid; DHA: docosahexaenoic acid; n3: omega 3; n6: omega 6; PUFA: polyunsaturated fatty acids). The microalgae isolates are stored in dark bottles at 2-6 0C and can last for minimum 6 months. Algal culture can be easily initiated by diluting the liquid extract or powder in transparent containers such as plastic bags or bottles followed by inoculating algal cells provided. Said microalgae culture medium can be used as solid medium 'for growing microalgal stocks by adding agar into it.
EPA DHA n3 n6 PUFA
Example 3
The filtered liquid extract is subjected to chemical analysis to determine its nutrient content. Table below shows the chemical and physical parameters of said filtered extract from shrimp ponds and Conway medium. Values are mean ± standard deviation.
Claims
1. A method for production of microalgae culture medium comprising the steps of: a) collecting of sludge;
b) extracting the interstitial water from said collected sludge to obtain the liquid concentrate; c) filtering said extracted liquid concentrate; d) assessing nutritional content of said filtered liquid concentrate; e) sterilizing said filtered liquid concentrate; and f) concentrating said sterile liquid concentrate to obtain the nutrient rich powder.
2. A method as claimed in claim 1, wherein said extracting the interstitial water is done by centrifugation or compression.
3. A method as claimed in claims 1 and 2, wherein said extraction is performed at the speed of 6000 to 8000 rpm for 20 to 40 minutes.
4. A method as claimed in claim 1 , wherein said filtering of said extracted liquid concentrate is glass fiber filtration and membrane filtration.
.
5. A method as claimed in claims 1 and 4, wherein said membrane filtration is done
under the pore size of 0.22 and 0.45 μm.
6. A method as claimed in claim 1, wherein said sterilizing of said filtered liquid concentrate is exposing said interstitial water to UV light for 1-2 hours.
7. A method as claimed in claim 1, wherein said concentrating of said sterile liquid concentrate further comprising the steps of: a) forming of ice from said sterile liquid concentrate; b) freeze-drying of said ice in freeze drier to obtain the nutrient rich powder.
8. A method as claimed in claim 7, wherein the freeze drying is done under the temperature of -40 to -55 0C and vacuum pressure of 130-150><103 mbar.
9. A method as claimed in claims 7 and 8, wherein all steps of said methods are conducted under sterile condition.
10. A liquid concentrate or nutrient rich powder produced from any of the preceding claims.
11. Use of the liquid concentrate or nutrient rich powder as claimed in claim 10 in the method for producing microalgae culture medium.
12. A method for producing microalgae culture medium by mixing Ig of said nutrient rich powder, as claimed in claim 10, to 5 to 10 liters of water.
13. A kit to initialize microalgae growth comprising: a) nutrient rich liquid concentrate and/or powder as claimed in claim 11 , and b) at least one type of microalgae isolates is supplemented with said nutrient rich liquid concentrate or powder.
14. A kit as claimed in claim 14, wherein said microalgae isolates are Chaetoceros spp. , Skeletonema spp., Isochrysis spp., Amphora spp., Cymbellq spp. Navicula spp., Chlorella spp., Nannochloropsis spp., Tetraselmis spp., Oscillatoria spp. Spirulina spp., and Pseudanabaena spp.
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2006800384486A CN101292023B (en) | 2005-10-17 | 2006-10-17 | Starter kit for the production of pure and high quality microalgae |
HK09102081.7A HK1122064A1 (en) | 2005-10-17 | 2009-03-04 | Starter kit for the production of pure and high quality microalgae |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
MYPI20054861 | 2005-10-17 | ||
MYPI20054861A MY159740A (en) | 2005-10-17 | 2005-10-17 | Starter kit for the production of pure and high quality microalgae |
Publications (1)
Publication Number | Publication Date |
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WO2007046685A1 true WO2007046685A1 (en) | 2007-04-26 |
Family
ID=37962737
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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PCT/MY2006/000019 WO2007046685A1 (en) | 2005-10-17 | 2006-10-17 | Starter kit for the production of pure and high quality microalgae |
Country Status (4)
Country | Link |
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CN (1) | CN101292023B (en) |
HK (1) | HK1122064A1 (en) |
MY (1) | MY159740A (en) |
WO (1) | WO2007046685A1 (en) |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2012005724A1 (en) | 2010-07-07 | 2012-01-12 | Universal Display Corporation | Host material for organic light emitting devices |
WO2012108877A1 (en) | 2011-02-11 | 2012-08-16 | University Display Corporation | Organic light emitting device and materials for use in same |
WO2012108879A1 (en) | 2011-02-11 | 2012-08-16 | Universal Display Corporation | Organic light emitting device and materials for use in same |
WO2012108878A1 (en) | 2011-02-11 | 2012-08-16 | Universal Display Corporation | Organic light emitting device and materials for use in same |
WO2012108881A1 (en) | 2011-02-11 | 2012-08-16 | Universal Display Corporation | Organic light emitting device and materials for use in same |
WO2014125429A1 (en) * | 2013-02-13 | 2014-08-21 | Centre For Cellular And Molecular Platforms | Culture medium for growth of recombinant proteins |
Citations (4)
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US4087936A (en) * | 1976-12-13 | 1978-05-09 | Mobil Oil Corporation | Process for production of alga biopolymer and biomass |
WO1994007993A1 (en) * | 1992-10-07 | 1994-04-14 | Conservatoire National Des Arts Et Metiers | Method of producing microalgae |
JP2000224981A (en) * | 1999-02-04 | 2000-08-15 | Marine Biotechnol Inst Co Ltd | Medium for marine microalgae and method for culturing marine microalgae using the medium |
KR20020050386A (en) * | 2000-12-21 | 2002-06-27 | 이구택 | Culturing method of micro-algae with high contents of fatty acids by using steelmaking slag |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
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CN1129001C (en) * | 2000-12-07 | 2003-11-26 | 中国科学院南京土壤研究所 | Method for analyzing water in gap between deposits |
-
2005
- 2005-10-17 MY MYPI20054861A patent/MY159740A/en unknown
-
2006
- 2006-10-17 CN CN2006800384486A patent/CN101292023B/en not_active Expired - Fee Related
- 2006-10-17 WO PCT/MY2006/000019 patent/WO2007046685A1/en active Application Filing
-
2009
- 2009-03-04 HK HK09102081.7A patent/HK1122064A1/en not_active IP Right Cessation
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4087936A (en) * | 1976-12-13 | 1978-05-09 | Mobil Oil Corporation | Process for production of alga biopolymer and biomass |
WO1994007993A1 (en) * | 1992-10-07 | 1994-04-14 | Conservatoire National Des Arts Et Metiers | Method of producing microalgae |
JP2000224981A (en) * | 1999-02-04 | 2000-08-15 | Marine Biotechnol Inst Co Ltd | Medium for marine microalgae and method for culturing marine microalgae using the medium |
KR20020050386A (en) * | 2000-12-21 | 2002-06-27 | 이구택 | Culturing method of micro-algae with high contents of fatty acids by using steelmaking slag |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2012005724A1 (en) | 2010-07-07 | 2012-01-12 | Universal Display Corporation | Host material for organic light emitting devices |
WO2012108877A1 (en) | 2011-02-11 | 2012-08-16 | University Display Corporation | Organic light emitting device and materials for use in same |
WO2012108879A1 (en) | 2011-02-11 | 2012-08-16 | Universal Display Corporation | Organic light emitting device and materials for use in same |
WO2012108878A1 (en) | 2011-02-11 | 2012-08-16 | Universal Display Corporation | Organic light emitting device and materials for use in same |
WO2012108881A1 (en) | 2011-02-11 | 2012-08-16 | Universal Display Corporation | Organic light emitting device and materials for use in same |
WO2014125429A1 (en) * | 2013-02-13 | 2014-08-21 | Centre For Cellular And Molecular Platforms | Culture medium for growth of recombinant proteins |
Also Published As
Publication number | Publication date |
---|---|
HK1122064A1 (en) | 2009-05-08 |
CN101292023B (en) | 2011-08-10 |
MY159740A (en) | 2017-01-31 |
CN101292023A (en) | 2008-10-22 |
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