CN104593261A - Chaetoceros minutissismus concentrated culture solution and culture method for culturing Chaetoceros minutissismus - Google Patents

Chaetoceros minutissismus concentrated culture solution and culture method for culturing Chaetoceros minutissismus Download PDF

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CN104593261A
CN104593261A CN201510002937.7A CN201510002937A CN104593261A CN 104593261 A CN104593261 A CN 104593261A CN 201510002937 A CN201510002937 A CN 201510002937A CN 104593261 A CN104593261 A CN 104593261A
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王培磊
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Linyi University
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    • 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
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Abstract

The invention discloses a preparation method of a Chaetoceros minutissismus concentrated culture solution and high-efficiency culture technology, belonging to the aquaculture field. The prepared Chaetoceros minutissismus concentrated culture solution can be stored and transported conveniently and can be produced conveniently in a factory-like, standard and specialty manner, microconstituents can be quantified accurately, and a traditional f/2 nutritive salt formula is optimized, and the constituents are reasonable and balanced and are beneficial to the rapid growth of the microalgae; a plastic purified water barrel is used for breeding Chaetoceros minutissismus, the operation is flexible, pollution cannot be caused easily, the washing operation is convenient, and the plastic purified water barrel cannot be scratched easily, and is low in cost; different illuminations and temperatures are provided in early, middle and later culture periods, nutritive salt is supplemented in the later culture period to promote the continuous and rapid growth of the Chaetoceros minutissismus, and the algal yield is improved by 120%; and three plastic barrels are connected in series for breeding, introduced gas is utilized for three times, and thus energy and resources are saved.

Description

A kind of small-sized Chaetoceros Concentrated culture fluids and cultivate the cultural method of small-sized Chaetoceros
Technical field the invention belongs to aquaculture field, particularly relates to a kind of small-sized Chaetoceros Concentrated culture fluids preparation method and High effecfive technique.
Background technology
Small-sized Chaetoceros Chaetoceros minutissismus is a kind of small ocean planktonic diatom, is subordinate to Bacillariophyta, center guiding principle, box-like Cutleriales, Chaetoceros section, and Chaetoceros belongs to.Cell is little, and wall is thin, most individual cells, also has 2-3 cell to be linked to be colony.Shell face is oval to circular, and central authorities are slightly protruding, and minority is smooth.Girdle face rectangularity is to quadrangle, and the long 4.6-9.2 micron of cell, wide 3.45-4.6 micron, girdle band is not obvious.Angle capillary and growing, end point, bears from cell corner, almost with longitudinal axis, long 20.7-34.5 micron.The angle hair at two ends is slightly serpentine centered by cell paste.Chromatoplast one, sheet, tawny.In culturing process, cell is often out of shape.The cell of distortion elongates, the angle hair completely dissolve in the shortening of angle hair or a shell face.After distortion, frond is larger than normal.Under normal circumstances, small-sized Chaetoceros nutrient solution is in golden yellow, and without precipitation, culture density can reach 330,000/ml, suitable salinity scope 26-35 ‰, thermophilic 5-30 DEG C, growth rate raises with water temperature and improves, the decline more than 30 DEG C, optimum temperuture 30 DEG C, the suitableeest illumination 10000-15000lux.Suitable p H6.4-9.5, the suitableeest 8.0-8.9.Binary fission is bred, and can form statospore, also can form auxospore when environment is bad.Belong to wide temperature, wide salt algae, water temperature 20-30 DEG C, salinity 30 ‰ time growth the fastest.With other Micro Algae as compared with Isochrysis galbana, station-service power source, flat algae, Phaeodactylum tricornutum, this algae propagation is fast, easily cultivates, high temperature resistant, being of high nutritive value, is the precious marine product high-quality nursery bait such as prawn, sea cucumber, clam, oyster, abalone, stalwart blood clam, sea urchin.
About micro algae culturing liquid, traditional method is matching while using, with how much joining how many, shortcoming is troublesome poeration, and workload is large, particularly some trace ingredientss and element, consumption is few, and weigh trouble, error is large, and nutrient solution inconvenience transport and commercialization, market operation, be not suitable with that the micro-algae of modern marine is extensive, intensive, the trend of factorial praluction.At present, the subject matter that small-sized Chaetoceros pilot scale culture exists is that frustule poor growth, doubling time are long.Past Yang Yan person of outstanding talent has invented culture medium prescription and total man's work fast culture process (application number CN201310420414) of a kind of Chaetoceros muelleri.Culture medium prescription comprises urea 0.1-0.3g/L, sodium bicarbonate 0.05-0.1g/L, water glass water crystallization 0.1-0.5g/L, potassium primary phosphate 0.01-0.02g/L, EDTA 3-5g/L, VB 11-5mg/L, VB 121-3 μ g/L, formula solvent is seawater; Cultural method is to after culture tank, seawater sterilization, be that solvent is according to joining substratum above with seawater, be equipped with in the culture tank of substratum by connecing the access of algae kind, cultivate after sealing, carry out the illumination of three primary colours light during cultivation and be filled with being less than 0.1 μm of air filtering core filtered air through aperture, cultivate and gather after 3-5 days.This invention culture medium prescription is balanced in nutrition, cost is low, and cultural method makes Chaetoceros grow fast, and process operation is simple, but culture tank volume is little, once can only cultivate a small amount of algae liquid, be difficult to apply aborning, and it is large to purchase culture tank investment, technical requirements is high, and common laborer is difficult to grasp.Yang Hailin has invented a kind of high-density culture technique (application number CN201010509643) of autotrophy oil-producing microalgae, adopts the fermentation process that cutting method and two step method are coupled, with the CO of power plant emission 2waste gas is carbon source, using the municipal wastewater after filtering and impurity removing bacterium or assorted algae as substratum, high-density culture oil-producing microalgae, micro-algae be selected from chlorella, micro-plan ball algae, Botryococcus braunii, Chaetoceros gracilis, extra large Chlorococcum, Isochrysis galbana, Nitzschia closterium minutissima, Phaeodactylum tricornutum, Dunaliella salina and rhombus algae one or more, this invention can realize nitrogen, phosphorus and CO 2high removal efficiency and concentration of algae, grease and protein content are high, reach protection of the environment and production Biological resources double effects, cost is low, pollution-free, and comprehensive benefit is high, but this method technical process is long, and troublesome poeration, easily pollutes, inefficiency.
In order to overcome the deficiency of conventional art, the invention discloses a kind of small-sized Chaetoceros Concentrated culture fluids preparation method and High effecfive technique.Technical superiority of the present invention shows: 1) abandoned the way of traditional nutrient solution matching while using, prepare small-sized Chaetoceros Concentrated culture fluids, easy to use, workload is little, storage, convenient transportation, be convenient to the batch production of nutrient solution, stdn, the marketization, commercialization, Virtual production, trace ingredients and element are quantitatively accurate, and error is little, and are optimized the f/2 nutritive salt formula that tradition uses, supplement chicken manure leach liquor, NaHCO 3, people urinates, Na 2siO 3, calcium superphosphate, soil extract, ooze extracts liquid, MgSO 4, Ca (NO 3) 2, CaCl 2, CH 3cOONa, CH 3cH 2cOONa, the compositions such as indolylacetic acid, composition science, reasonable, balanced more, be conducive to micro-algae and grow fast and sterilize thoroughly, clean hygiene, not easily pollutes; 2) use plastics pure water barrel to cultivate small-sized Chaetoceros, flexible operation, not easily pollute, scrub conveniently, not easily scratch and break up, cost is low, and output is high; 3) growth rhythm of small-sized Chaetoceros is respected, according to the particular requirement of algae Different growth phases to envrionment conditions, different light application time, intensity and temperature is provided in the early, middle and late phase of cultivating, light application time is shorter in early days, intensity is less, avoid high light to cause light to injure to frustule, stronger intensity of illumination and time and higher temperature are provided late period, are conducive to the high unsaturated fatty acid such as EPA, DHA in cell and accumulate in a large number; And cultivate middle and later periods extra-nutrition salt, promote Chaetoceros continue, grow fast; Small-sized Chaetoceros output can improve 120%; 4) adopt three plastic tank series connection cultivation, the gas passed into obtains three times and utilizes, and has saved energy and resource.
Summary of the invention
In order to overcome the deficiency of current technology, the invention provides a kind of small-sized Chaetoceros Concentrated culture fluids and cultivate the cultural method of small-sized Chaetoceros, concrete grammar is as follows:
The preparation of the small-sized Chaetoceros Concentrated culture fluids of step 1
Nutrient solution component prescription: chicken manure leach liquor 1 part, NaHCO 30.6 part, people urinates 0.7 part, Na 2siO 30.02 part, f/2 trace element solution 0.8 part, f/2 vitamin solution 0.8 part, calcium superphosphate 0.1 part, soil extract 0.6 part, ooze extracts liquid 0.8 part, MgSO47H 2o 0.2 part, Ca (NO 3) 20.3 part, CaCl 22H 2o 0.5 part, CH 3cOONa 0.05 part, CH 3cH 2cOONa 0.1 part, indolylacetic acid 0.001 part, boils refrigerated sea water 100 parts; Load Erlenmeyer flask, be placed on shaking table and shake up 15 minutes, prepared by Concentrated culture fluids; Wherein f/2 trace element solution formula (by preparation 1000 ml soln): ZnSO 44H 2o 23 milligrams, MnCl 24H 2o 178 milligrams, CuSO 45H 2o 10 milligrams, FeC 6h 5o 75H 2o 3.9 grams, Na 2moO 42H 2o 7.3 milligrams, CoCl 26H 2o 12 milligrams, Na 2eDTA 4.35 grams, distilled water 1000 milliliters; F/2 vitamin solution formula (by preparation 1000 ml soln): vitamins B 120.5 milligram, vitamins B 1100 milligrams, vitamin H 0.5 milligram, distilled water 1000 milliliters.
Step 2 small-sized Chaetoceros high-efficient culture method
Get the plastics pure water barrel (commercially available) of the band handle of three 20 liters, add nutrient solution described in step 1 1000 milliliters respectively, then add the ordinary sea water that 9000 milliliters are boiled cooling, shake up; Choosing growth small-sized Chaetoceros algae kind that is vigorous, pollution-free, that be in exponential phase of growth is inoculated, and inoculum density 40,000 cells/ml algae liquid, shake up 3 minutes gently.Each pure water barrel bottleneck inserts two Glass tubings, the diameter of pipe is 8 millimeters, one is gas-filled valve, another root is vapor pipe, the gas-filled valve of the 2nd bucket is connected with the vapor pipe of first bucket, the gas-filled valve of the 3rd bucket is connected with the vapor pipe of the 2nd bucket, and the gas be filled with like this, successively through three plastic tanks, can obtain 3 times and utilize.The gas-filled valve upper end of first bucket connects air compressor, and the other end is inserted into bottom algae liquid, is filled with mixed gas, gaseous constituent=pure CO of air 99%+ 20.5% ten nitrogen protoxide 0.3%+ ammonia 0.2%, 24 hours every days are inflation uninterruptedly, and tolerance should not be too large, can not float with frustule, these 3 plastic tanks are put into microdisk electrode room cultivate, room conditioning temperature control, fluorescent tube throws light on, and carrys out controlled light intensity with the quantity opening fluorescent tube, cultured continuously 9 days, within first 3 days, adjust the temperature to 25 DEG C, intensity of illumination 6000lux, light dark period=6h/L+6h/D, namely intermittent light is provided, continuous illumination in 6 hours, 6 hours continuous darknesses, so circulate, 4-6 days, adjust the temperature to 26 DEG C, intensity of illumination 9000lux, continuous illumination in 15 hours is provided every day, then 9 hours continuous darknesses, circulation like this, to the 5th day, the Concentrated culture fluids 50 milliliters described in step 1 added by each bottle, 7-9 days, adjust the temperature to 28 DEG C, intensity of illumination 12000lux, continuous illumination 24 hours every days, there is no dark, 7th day, distilled water 250 milliliters added by each bucket, to the 8th day, the Concentrated culture fluids 30 milliliters described in step 1 added by each bucket, by the 9th day, small-sized Chaetoceros cell density can reach 750,000/milliliter algae liquid, cultivate complete.
Beneficial outcomes
About micro algae culturing liquid, traditional method is matching while using, and with how much joining how many, troublesome poeration, workload is large, particularly some trace ingredientss and element, and consumption is few, weighs trouble, and error is large, and inconvenience transport and commercialization, market operation.In order to overcome the deficiency of conventional art, the invention discloses a kind of small-sized Chaetoceros Concentrated culture fluids preparation method and High effecfive technique.This technical superiority shows: 1) abandoned the way of traditional nutrient solution matching while using, prepare small-sized Chaetoceros Concentrated culture fluids, easy to use, workload is little, storage, convenient transportation, be convenient to the batch production of nutrient solution, stdn, the marketization, commercialization, Virtual production, trace ingredients and element are quantitatively accurate, and error is little, and are optimized the f/2 nutritive salt formula that tradition uses, composition science, reasonable, balanced more, is conducive to micro-algae and grows fast; 2) use plastics pure water barrel to cultivate small-sized Chaetoceros, light transmission is good, and flexible operation not easily pollutes, and scrub conveniently, not easily scratch and break up, cost is low, and output is high; 3) growth rhythm of small-sized Chaetoceros is respected, according to the particular requirement of algae Different growth phases to envrionment conditions, there is provided suitable light application time, intensity and temperature in the early, middle and late phase of cultivating, to be conducive in cell the high unsaturated fatty acids such as EPA, DHA and to accumulate in a large number; And at cultivation middle and later periods extra-nutrition salt, promote that Chaetoceros continues, grows fast; Small-sized Chaetoceros output can improve 120%; 4) adopt three plastic tank series connection cultivation, the gas passed into obtains three times and utilizes, and has saved energy and resource.
Accompanying drawing explanation
Fig. 1 is small-sized Chaetoceros form;
Fig. 2 is small-sized Chaetoceros high-efficient culture method, and wherein Reference numeral is: 1 gas-filled valve, 2 vapor pipes; Arrow indication is gas trend.
Embodiment
Small-sized Chaetoceros Concentrated culture fluids and cultivate the cultural method of small-sized Chaetoceros, concrete grammar is as follows:
The preparation of the small-sized Chaetoceros Concentrated culture fluids of step 1
Chicken manure leach liquor 1 part, NaHCO 30.6 part, people urinates 0.7 part, Na 2siO 30.02 part, f/2 trace element solution 0.8 part, f/2 vitamin solution 0.8 part, calcium superphosphate 0.1 part, soil extract 0.6 part, ooze extracts liquid 0.8 part, MgSO47H 2o0.2 part, Ca (NO 3) 20.3 part, CaCl 22H 2o 0.5 part, CH 3cOONa 0.05 part, CH 3cH 2cOONa 0.1 part, indolylacetic acid 0.001 part, boils refrigerated sea water 100 parts; Load Erlenmeyer flask, be placed on shaking table and shake up 15 minutes, prepared by Concentrated culture fluids; Wherein f/2 trace element solution formula (by preparation 1000 ml soln): ZnSO 44H 2o 23 milligrams, MnCl 24H 2o 178 milligrams, CuSO 45H 2o 10 milligrams, FeC 6h 5o 75H 2o 3.9 grams, Na 2moO 42H 2o 7.3 milligrams, CoCl 26H 2o 12 milligrams, Na 2eDTA 4.35 grams, distilled water 1000 milliliters; F/2 vitamin solution formula (by preparation 1000 ml soln): vitamins B 120.5 milligram, vitamins B 1100 milligrams, vitamin H 0.5 milligram, distilled water 1000 milliliters.
Step 2 small-sized Chaetoceros high-efficient culture method
Get the plastics pure water barrel (commercially available) of the band handle of three 20 liters, add small-sized Chaetoceros Concentrated culture fluids 1000 milliliters according to claim 1 respectively, then add the ordinary sea water that 9000 milliliters are boiled cooling, shake up, choosing growth small-sized Chaetoceros algae kind that is vigorous, pollution-free, that be in exponential phase of growth is inoculated, and inoculum density 40,000 cells/ml algae liquid, shake up 3 minutes gently, each pure water barrel bottleneck inserts two Glass tubings, the diameter of pipe is 8 millimeters, one is gas-filled valve, another root is vapor pipe, the gas-filled valve of the 2nd bucket is connected with the vapor pipe of first bucket, the gas-filled valve of the 3rd bucket is connected with the vapor pipe of the 2nd bucket, and the gas be filled with like this, successively through three plastic tanks, can obtain 3 times and utilize, the gas-filled valve upper end of first bucket connects air compressor, and the other end is inserted into bottom algae liquid, is filled with mixed gas, gaseous constituent=pure CO of air 99%+ 20.5%+ nitrogen protoxide 0.3%+ ammonia 0.2%, 24 hours every days are inflation uninterruptedly, and tolerance should not be too large, can not float with frustule, these 3 plastic tanks are put into microdisk electrode room cultivate, room conditioning temperature control, fluorescent tube throws light on, and carrys out controlled light intensity with the quantity opening fluorescent tube, cultured continuously 9 days, within first 3 days, adjust the temperature to 25 DEG C, intensity of illumination 6000lux, light dark period=6h/L+6h/D, namely intermittent light is provided, continuous illumination in 6 hours, 6 hours continuous darknesses, so circulate, 4-6 days, adjust the temperature to 26 DEG C, intensity of illumination 90001ux, continuous illumination in 15 hours is provided every day, then 9 hours continuous darknesses, circulation like this, to the 5th day, the Concentrated culture fluids 50 milliliters described in step 1 added by each bottle, 7-9 days, adjust the temperature to 28 DEG C, intensity of illumination 12000lux, continuous illumination 24 hours every days, there is no dark, 7th day, distilled water 250 milliliters added by each bucket, to the 8th day, the Concentrated culture fluids 30 milliliters described in step 1 added by each bucket, by the 9th day, small-sized Chaetoceros cell density can reach 750,000/milliliter algae liquid, cultivate complete.
In step 1, various nutritive salt will add by the order provided in formula, to avoid chemical reaction;
In step 1, various inorganic nutrient salt and VITAMIN can be made into mother liquor in advance and be placed in 4 DEG C of refrigerator cold-storages and save backup, and do not exceed 30 days storage period;
The preparation method of soil extract described in step 1: get the woods 1000 parts, soil at the middle and upper levels, adding distil water 1000 parts, then add NaOH 2.5 parts, heating, boils 45 minutes, cooling, and precipitate 12 hours, get supernatant liquor, filter paper filtering, filtrate is soil extract;
The leach liquor of chicken manure described in step 1 preparation method: dried poultrymanure, green grass, soya-bean cake, rice bran, the part by weight mixing of 60: 10: 12: 15: 2: 1 pressed by grain husk shell (shells of five cereals) and vegetable mould, stir, load airtight in ceramic cylinder fermentation, after 30 days, pour out, at clean water mire, stand lamellar, dry 48 hours, after get 2 kilograms, be placed in plastic bucket, add 10 liters, tap water, stir, with 80 order silk cover filterings, clear liquid is boiled 5 minutes, cooling, leave standstill 24 hours, get supernatant liquor, load reagent bottle, labelled, be placed in 4 DEG C of refrigerators, for subsequent use,
Ooze described in step 1 extracts liquid and preparation method thereof: get the more fertile ooze that what is not black again, add 2 parts of fresh water, stir with 1 part of mud, leave standstill 1-2 minute, get upper strata mud in aluminum pot, the amount adding 1 gram of NaOH by every 1000 milliliters of mud adds NaOH, boils 25 minutes.Constantly need stir when boiling, boil and leave standstill that within 24 hours, to get supernatant liquor for subsequent use afterwards;
Because of FeC in step 1 6h 5o 75H 2o (ironic citrate) comparatively indissoluble solution, can add a small amount of tap water low-grade fever on stove and, to 80-90 DEG C, and constantly be stirred to whole thawing;
In step 1, people urinates preparation method: get people and urinate 1000 milliliters, it is for subsequent use that Erlenmeyer flask boils rear cooling;
NaHCO described in step 1 3, Na 2siO 3, calcium superphosphate, MgSO 4, Ca (NO 3) 2, CaCl 2, CH 3cOONa, CH 3cH 2cOONa, the pharmaceutical chemicals purity such as indolylacetic acid all need analytical pure rank;
The small-sized Chaetoceros inoculated in step 2 carries out purifying in advance (this area ordinary method, as long as can realize purifying, such as Michaelis extraction method), and during to guarantee inoculation, algae kind is purebred, pollution-free and is in eugonic exponential phase of growth;
In step 2, plastic tank used adds 450 ml distilled water rinse 3 times after need washing, and bottle mouth position 75% alcohol wipes with sterilization;
In step 2, pure water barrel used should be transparent or white bucket, to increase light transmission;
Bung place scotch tape or glass cement sealing in step 2, with anti-gas-leak; Bucket is connected with plastics available between bucket or rubber mass pipe.
Last it is noted that above embodiment is only in order to illustrate technical scheme of the present invention, be not intended to limit; Although with reference to previous embodiment to invention has been detailed description, those of ordinary skill in the art is to be understood that: it still can be modified to the technical scheme described in previous embodiment, or carries out equivalent replacement to wherein portion of techniques feature; And these amendments or replacement, do not make the essence of appropriate technical solution depart from the spirit and scope of embodiment of the present invention technical scheme.

Claims (2)

1. a small-sized Chaetoceros Concentrated culture fluids, is characterized in that concrete component prescription is as follows:
Chicken manure leach liquor 1 part, NaHCO 30.6 part, people urinates 0.7 part, Na 2siO 30.02 part, f/2 trace element solution 0.8 part, f/2 vitamin solution 0.8 part, calcium superphosphate 0.1 part, soil extract 0.6 part, ooze extracts liquid 0.8 part, MgSO47H 2o0.2 part, Ca (NO 3) 20.3 part, CaCl 22H 2o 0.5 part, CH 3cOONa 0.05 part, CH 3cH 2cOONa 0.1 part, indolylacetic acid 0.001 part, boils refrigerated sea water 100 parts; Load Erlenmeyer flask, be placed on shaking table and shake up 15 minutes, prepared by Concentrated culture fluids; Wherein f/2 trace element solution formula (by preparation 1000 ml soln): ZnSO 44H 2o 23 milligrams, MnCl 24H 2o 178 milligrams, CuSO 45H 2o 10 milligrams, FeC 6h 5o 75H 2o 3.9 grams, Na 2moO 42H 2o 7.3 milligrams, CoCl 26H 2o 12 milligrams, Na 2eDTA 4.35 grams, distilled water 1000 milliliters; F/2 vitamin solution formula (by preparation 1000 ml soln): vitamins B 120.5 milligram, vitamins B 1100 milligrams, vitamin H 0.5 milligram, distilled water 1000 milliliters.
2. a kind of small-sized Chaetoceros Concentrated culture fluids as claimed in claim 1 cultivates the cultural method of small-sized Chaetoceros, it is characterized in that:
Get the plastics pure water barrel (commercially available) of the band handle of three 20 liters, add small-sized Chaetoceros Concentrated culture fluids 1000 milliliters according to claim 1 respectively, then add the ordinary sea water that 9000 milliliters are boiled cooling, shake up, choosing growth small-sized Chaetoceros algae kind that is vigorous, pollution-free, that be in exponential phase of growth is inoculated, and inoculum density 40,000 cells/ml algae liquid, shake up 3 minutes gently, each pure water barrel bottleneck inserts two Glass tubings, the diameter of pipe is 8 millimeters, one is gas-filled valve, another root is vapor pipe, the gas-filled valve of the 2nd bucket is connected with the vapor pipe of first bucket, the gas-filled valve of the 3rd bucket is connected with the vapor pipe of the 2nd bucket, and the gas be filled with like this, successively through three plastic tanks, can obtain 3 times and utilize, the gas-filled valve upper end of first bucket connects air compressor, and the other end is inserted into bottom algae liquid, is filled with mixed gas, gaseous constituent=pure CO of air 99%+ 20.5%+ nitrogen protoxide 0.3%+ ammonia 0.2%, 24 hours every days are inflation uninterruptedly, and tolerance should not be too large, can not float with frustule, these 3 plastic tanks are put into microdisk electrode room cultivate, room conditioning temperature control, fluorescent tube throws light on, and carrys out controlled light intensity with the quantity opening fluorescent tube, cultured continuously 9 days, within first 3 days, adjust the temperature to 25 DEG C, intensity of illumination 6000lux, light dark period=6h/L+6h/D, namely intermittent light is provided, continuous illumination in 6 hours, 6 hours continuous darknesses, so circulate, 4-6 days, adjust the temperature to 26 DEG C, intensity of illumination 9000lux, continuous illumination in 15 hours is provided every day, then 9 hours continuous darknesses, circulation like this, to the 5th day, the Concentrated culture fluids 50 milliliters described in step 1 added by each bottle, 7-9 days, adjust the temperature to 28 DEG C, intensity of illumination 12000lux, continuous illumination 24 hours every days, there is no dark, 7th day, distilled water 250 milliliters added by each bucket, to the 8th day, the Concentrated culture fluids 30 milliliters described in step 1 added by each bucket, by the 9th day, small-sized Chaetoceros cell density can reach 750,000/milliliter algae liquid, cultivate complete.
CN201510002937.7A 2015-01-06 2015-01-06 Chaetoceros minutissismus concentrated culture solution and culture method for culturing Chaetoceros minutissismus Pending CN104593261A (en)

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Publication number Priority date Publication date Assignee Title
CN105936877A (en) * 2016-03-07 2016-09-14 临沂大学 Method for culturing chaetoceros gracilis by using waste beer bottle
CN108485983A (en) * 2018-06-04 2018-09-04 海南源泉生物科技有限公司 A kind of culture medium and its cultural method for cultivating green alga for scale high density
CN108485983B (en) * 2018-06-04 2021-07-30 海南源泉生物科技有限公司 Culture medium for large-scale high-density cultivation of green algae and culture method thereof

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