CN105340792B - A kind of method and its application for screening intertidal shellfish seedling Mirco algal food - Google Patents
A kind of method and its application for screening intertidal shellfish seedling Mirco algal food Download PDFInfo
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- 235000015170 shellfish Nutrition 0.000 title claims abstract description 66
- 238000000034 method Methods 0.000 title claims abstract description 26
- 235000013305 food Nutrition 0.000 title claims abstract description 16
- 238000012216 screening Methods 0.000 title claims abstract description 10
- 229930182558 Sterol Natural products 0.000 claims abstract description 21
- 235000003702 sterols Nutrition 0.000 claims abstract description 21
- 239000000203 mixture Substances 0.000 claims abstract description 19
- 239000002253 acid Substances 0.000 claims abstract description 15
- 238000002156 mixing Methods 0.000 claims abstract description 14
- 150000003432 sterols Chemical class 0.000 claims abstract description 13
- 230000012010 growth Effects 0.000 claims abstract description 8
- 241001339782 Scapharca broughtonii Species 0.000 claims description 21
- 238000000513 principal component analysis Methods 0.000 claims description 18
- 125000001931 aliphatic group Chemical group 0.000 claims description 13
- 241000227752 Chaetoceros Species 0.000 claims description 10
- 241000206751 Chrysophyceae Species 0.000 claims description 9
- 238000002474 experimental method Methods 0.000 claims description 7
- 239000013535 sea water Substances 0.000 claims description 4
- 210000002784 stomach Anatomy 0.000 claims description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 2
- 238000004445 quantitative analysis Methods 0.000 claims description 2
- 150000003431 steroids Chemical class 0.000 claims description 2
- 235000014113 dietary fatty acids Nutrition 0.000 abstract description 9
- 239000000194 fatty acid Substances 0.000 abstract description 9
- 229930195729 fatty acid Natural products 0.000 abstract description 9
- -1 fatty acid sterols Chemical class 0.000 abstract description 9
- 230000000694 effects Effects 0.000 abstract description 7
- 210000000577 adipose tissue Anatomy 0.000 abstract description 2
- 238000013461 design Methods 0.000 abstract description 2
- 241000195493 Cryptophyta Species 0.000 description 12
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- 238000004458 analytical method Methods 0.000 description 6
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- 241000386616 Pavlova viridis Species 0.000 description 5
- 235000020639 clam Nutrition 0.000 description 5
- VLKZOEOYAKHREP-UHFFFAOYSA-N hexane Substances CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241000091752 Chaetoceros calcitrans Species 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 238000001816 cooling Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000007789 sealing Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 2
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 235000003642 hunger Nutrition 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
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- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
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- 230000037351 starvation Effects 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 241000206761 Bacillariophyta Species 0.000 description 1
- 206010013786 Dry skin Diseases 0.000 description 1
- 241000224472 Eustigmatophyceae Species 0.000 description 1
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-UHFFFAOYSA-N 0.000 description 1
- 241001501885 Isochrysis Species 0.000 description 1
- 241000278693 Moerella Species 0.000 description 1
- 241000237852 Mollusca Species 0.000 description 1
- XCOBLONWWXQEBS-KPKJPENVSA-N N,O-bis(trimethylsilyl)trifluoroacetamide Chemical compound C[Si](C)(C)O\C(C(F)(F)F)=N\[Si](C)(C)C XCOBLONWWXQEBS-KPKJPENVSA-N 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 239000004115 Sodium Silicate Substances 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical class [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 229920006221 acetate fiber Polymers 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229960001948 caffeine Drugs 0.000 description 1
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
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- 230000007812 deficiency Effects 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 230000009246 food effect Effects 0.000 description 1
- 235000021471 food effect Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
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- 230000000366 juvenile effect Effects 0.000 description 1
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- 230000009571 larval growth Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229910052603 melanterite Inorganic materials 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 238000002705 metabolomic analysis Methods 0.000 description 1
- 230000001431 metabolomic effect Effects 0.000 description 1
- JBXYCUKPDAAYAS-UHFFFAOYSA-N methanol;trifluoroborane Chemical compound OC.FB(F)F JBXYCUKPDAAYAS-UHFFFAOYSA-N 0.000 description 1
- BABMCXWQNSQAOC-UHFFFAOYSA-M methylmercury chloride Chemical compound C[Hg]Cl BABMCXWQNSQAOC-UHFFFAOYSA-M 0.000 description 1
- 231100000207 neurotoxicity testing Toxicity 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Inorganic materials [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000027272 reproductive process Effects 0.000 description 1
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- 239000004576 sand Substances 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 235000019795 sodium metasilicate Nutrition 0.000 description 1
- NTHWMYGWWRZVTN-UHFFFAOYSA-N sodium silicate Chemical compound [Na+].[Na+].[O-][Si]([O-])=O NTHWMYGWWRZVTN-UHFFFAOYSA-N 0.000 description 1
- 229910052911 sodium silicate Inorganic materials 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 125000002328 sterol group Chemical group 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Farming Of Fish And Shellfish (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The present invention provides a kind of method for screening intertidal shellfish seedling Mirco algal food, i.e., is ingested the composition of shellfish body fat acid and sterol after mixing microalgae by quantitatively detecting the intertidal shellfish young, carry out the accurate method for screening the high-quality Mirco algal food of intertidal shellfish seedling.The cultivation cycle of the method for the present invention is short, aquatic environment condition is avoided on the issuable influence of culture effect, the microalgae filtered out based on shellfish larvae fatty acid sterols composition, the design choices of shellfish larvae and assimilation in the more realistic growth course of its bait effect, so as to the selection as the high-quality bait of shellfish larvae.
Description
Technical field
The invention belongs to aquaculture bait culture technique field, and in particular to one kind screening intertidal shellfish seedling microalgae bait
The method of material.
Background technology
The intertidal shellfish yield such as China " clam, blood clam, razor clam " accounts for marine shellfish cultivation total output nearly 40% at present.And in beach shellfish
In class nursery, the kind of Mirco algal food directly decides the success or failure of nursery.Long-term practice is found, is fed in seedling raising process different micro-
During algae, intertidal shellfish larval growth speed difference is very big, illustrates that microalgae species has directly to growing for intertidal shellfish seedling
Connect influence.In actual intertidal shellfish seedling reproductive process, it would be desirable to which there is obvious bait to imitate to the intertidal shellfish young for selection
The Mirco algal food of fruit carries out expanding numerous feed.
Certain bait micro-algae to intertidal shellfish young bait effect how, it is typically all not of the same race by being fed to intertidal shellfish
The microalgae of class, judge the bait effect of corresponding microalgae by observing the speed of growth and the survival rate of corresponding shellfish seedling【Zhu Yu
Influence auspicious etc., that 2010.5 kinds of microalgaes grow to 4 kinds of beach juvenile mollusks, oceanographic research, 28 (3):60-66】.But the means need
Long cultivation cycle, in addition, condition of water quality, environmental condition can have an impact to the speed of growth and survival rate of seedling.
So generally require finally determine bait effect of the different microalgae to shellfish by multiple repetition test.Therefore, just need
A kind of method of effective detection microalgae as shellfish bait is provided.It is moreover, generally micro- using two or more are fed at present
The mixing of algae feeds mode, and this mode needs to cultivate different microalgaes, causes the cost increase of bait.
The content of the invention
The present invention provides a kind of method for screening intertidal shellfish seedling Mirco algal food, i.e., by quantitatively detecting intertidal shellfish children
Body ingest mixing microalgae after shellfish body fat acid and sterol composition, come accurately screen the high-quality Mirco algal food of intertidal shellfish seedling
Method, so as to make up the deficiencies in the prior art.
The method of the present invention, including the steps：
1) acquisition of standard group PCA shot charts
Feed shellfish larvae as standard group by the use of mixing microalgae, feed aliphatic acid that shellfish larvae is determined after some cycles and
Sterol forms, and then carries out principal component analysis (PCA) to aliphatic acid and sterol composition, obtains PCA shot charts；
2) under same cultivating condition, feed different types of single microalgae respectively to shellfish larvae and carry out culture conduct
Experimental group, the aliphatic acid and sterol composition of different experiments group shellfish larvae are determined after same period is fed, then to aliphatic acid
Formed with sterol and carry out principal component analysis, filtered out according to distribution of each experimental group on PCA shot charts close with standard group
Bait of the microalgae of experimental group as the shellfish.
Described shellfish is the shellfish of mudflat aquaculture, such as mud blood clam, blood clam, clam, razor clam of hanging, moerella irideseens, clam etc..
For microalgae used in the present invention in positioned at exponential phase of growth, concentration reaches 80~120cell μ l-1Shi Jinhang is thrown
Hello；
The shellfish of the present invention is fed before Mirco algal food, it is necessary to be substantially free of the clean natural sea of any microalgae by husky filter
Not bait throwing in starvation 8-10h is with residual bait in emptying stomach as far as possible in water.
The present invention carries out the quantitative analysis of aliphatic acid and sterol using gas chromatography-mass spectrography.
Using the method for the present invention, the bait micro-algae for determining mud blood clam is Chaetoceros or chrysophyceae.
Compared with prior art, the advantage of the invention is that：Cultivation cycle is short, avoids aquatic environment condition and culture is imitated
The issuable influence of fruit, the microalgae filtered out based on shellfish larvae fatty acid sterols composition, its bait effect are more realistic
The design choices of shellfish larvae and assimilation in growth course, so as to the selection as the high-quality bait of shellfish larvae.
Brief description of the drawings
Fig. 1:First based on mud blood clam seedling fatty acid sterols principal component analysis and the 3rd principal component scores figure (A) and
Two and the 3rd principal component scores figure (B)
Sample is respectively that single feeds P.viridis mud blood clam seedling (IG-PV), single feeds C. on figure
Calcitrans (TG-CC), I.galbana (TG-IG) and N.oculata (NO) mud blood clam seedling；Mixing is fed two-by-two
I.galbana and N.oculata (TG-IG-NO), I.galbana and P.viridis (TG-IG-NO), C.calcitrans and
I.galbana (TG-CC-IG), C.calcitrans and P.viridis (TG-CC-PV), C.calcitrans and
N.oculata (TG-CC-NO) and all 4 kinds of microalgaes mix the mud blood clam seedling (TG-MIX) fed entirely.
Embodiment
The present invention to the shellfish larvae for feeding mixing Mirco algal food and single Mirco algal food by determining fatty acid sterols group
Into, and carry out PCA analyses, on PCA shot charts, if fed the shellfish larvae sample after certain microalgae fed with mixing it is micro-
The shellfish larvae sample of algae bait is relatively, it is possible to illustrates that the microalgae is preferentially ingested and assimilated by the shellfish, actual shellfish
High-quality bait of the algae as the shellfish larvae is may be selected by during class larval culture.The present invention applies the technology, significantly
Incubation very long in conventional art is shortened, and avoids aquatic environment condition to the issuable influence of culture effect,
The assimilation selection of ingesting of shellfish larvae during the Mirco algal food effect that filters out is more realistic.
Principal component analytical method (principal components analysis, PCA) used in the present invention is a kind of
Multivariate data statistical method, the PCA shot charts of material clustering distribution are shown on two-dimensional space, can sample room material directly perceived
The similar or non-similarity of composition【Vliet E V et al.,2008.A novel in vitro metabolomics
approach for neurotoxicity testing,proof of principle for methyl mercury
chloride and caffeine.NeuroToxicology,29(1):1–12】。
Described mixing microalgae is completed after the various microalgaes for the shellfish that will have been reported mix.
The present invention is described in further detail by the following examples.
Embodiment 1
In September, 2014 often with 4 kinds of marine microalgaes, is divided in Bao Zhi Aqua Sciences Inc. of Fujian Province, selection intertidal shellfish
It is not：Chaetoceros (Chaetoceros calcitrans), the Isochrysis galbana (Isochrysis of Chrysophyta of Bacillariophyta
) and pavlova viridis (Pavlova viridis), the Nannochloropsis oculata of Eustigmatophyceae algae doors galbana
(Nannochloropsis oculata) is cultivated, and the culture seawater (salinity 20) of microalgae is filtered through 0.45 μm of acetate fiber
After membrane filtration, cooling is boiled.Add NMB3Nutrient solution (100mgL-1KNO3, 10mg·L-1KH2PO4,0.25mg·L- 1MnSO4·H2O,2.50mg·L-1FeSO4·7H2O,10mg·L-1 EDTA-Na2,6μg·L-1VB1,0.05μg·L-1VB12),
Add sodium metasilicate (20mg.L simultaneously-1) silicon source as Chaetoceros.Microalgae algae kind [natural light, training in 2500mL conical flask
Support temperature be (20 ± 2) DEG C] cultivate 1 week after, they are transferred in the 50L white bucket of plastics, natural light, natural temperature (25~
29 DEG C) under be further cultured for 1 week or so, when microalgae density reaches 80~120cell μ l-1When feed and give shellfish seedling.
The sand that salinity is 20 is separately added into the white plastic basin (experiment basin) that length is respectively 25,15 and 6cm
Seawater is filtered, and adds the thick 200 DEG C of dried disinfectings of 1-2mm and the filtered fresh ooze of nylon bolting silk through 75 μm of mesh, will be wet
Weight is that 0.68 ± 0.05mm of 1.5g average lengths mud blood clam children shellfish sample is uniformly spilled into 10 groups of experiment basins (every group parallel 3 respectively
It is individual), it is each to test in basin respectively at 08:00 and 20:It is micro- that 00 or so moment launched single microalgae or mixing in enough corresponding tables 1
Algae (equal proportion mixing), after being cultivated 7 days at 25.2-31.7 DEG C of natural temperature, sample is collected, it is natural in the cleaning by husky filter
Not bait throwing in starvation 8-10h empties residual bait in stomach in seawater, and fatty acid sterols analysis, while grab sample are carried out after freeze-drying
Measure the growth (table 2) of 30 young shellfishes.
Table 1 feeds the food species selected in experiment
The analysis of mud blood clam children shellfish fatty acid sterols comprises the following steps that：Take the 0.2g mud blood clams after crushing that are freeze-dried and mill
Young shellfish sample, with reference to Bligh-Dyer methods [Bligh E G, Dyer the W J.1959.A Rapid Method after improvement
Lipid Extraction and Purification,Can J Biochem Physiol,37:911-917] the total fat of extraction；
2mL 5-6%KOH methanol-waters (V/V=4 is added after total fat extraction:1) solution, inflated with nitrogen 1min, sealing, 60 DEG C of water-bath saponification
2h, cooling, saponification liquor add 15mL saturated nacl aqueous solution 1mL, then with HCl (1:1) pH is adjusted to be less than 1, chloroform：N-hexane (V/V
=1:4) 6mL is extracted in three times, after rotavapor under vacuum is dried, adds 0.5mL 14%BF3-CH3OH solution, inflated with nitrogen
1min, sealing, 60 DEG C of water-bath esterification 1h, cooling, with 6mL points of 3 extractions of n-hexane, merge extract solution, with 2mL redistilled waters
Wash once, upper liquid removes, and adds 1g anhydrous sodium sulfates, and absorb water 12h, pipettes upper liquid, N2100 μ L excess is added after drying
BSTFA, sealing, 60 DEG C of water-bath 2h, N2 dryings, n-hexane constant volume, carry out GC-MS analyses.GC conditions：250 DEG C of injector temperature,
Carrier gas is high-pure helium, the mL/min of column flow rate 0.81, and 73.0Kpa is pressed before post, 150 DEG C of post initial temperature, 3.5min is kept, with 20
DEG C/min rises to 200 DEG C, 5min is kept, then 280 DEG C/min is risen to 5 DEG C/min, keep 30min.Shunted when being analyzed with EI sources
Enter 1 μ L, split ratio 50:1.MS conditions：Analyzed with electron bombardment ionization source (EI), electron energy 70eV, ion source temperature 200
DEG C, 250 DEG C of interface temperature, choose SCAN patterns, mass scan range 40-600, solvent delay 3.5min.Finally according to GC-
The fragment ion quality collection of illustrative plates of each component in EIMS-TIC, by NIST storehouses and WILEY library searchings and with reference to aliphatic acid and steroid
Alcohol standard spectrogram and bibliography are identified.Percentage composition (the table of aliphatic acid and sterol constituent is calculated with area normalization method
2, represented with accounting for the percentage i.e. percentage contents of aliphatic acid and sterol total amount).
Table 2：Feed the fatty acid sterols composition (%) of mud blood clam children shellfish after different bait
By the measurement result input SIMCA-P-12.0 analysis softwares (Umetrics AB companies of Sweden) of fatty acid sterols
Carry out principal component analysis.(Fig. 1) can be observed on the shot chart of software output, and mixing feeds chrysophyceae with the green balls of Ba Fuzao or micro-
The sample (TG-IG-PV and TG-IG-NO on figure) of algae approaches with individually feeding chrysophyceae (TG-IG figure on), and is fed bar with independent
The sample of husband algae (TG-PV on figure) or Nannochloropsis oculata (TG-NO on figure) is far apart；Equally, mixing feeds Chaetoceros with a bar husband
The sample of algae or Nannochloropsis oculata (TG-CC-PV and TG-CC-NO on figure) is close with individually feeding Chaetoceros (TG-CC on figure), and
Sample with individually feeding Ba Fuzao (TG-PV on figure) or Nannochloropsis oculata (TG-NO on figure) is far apart；And mix and feed chrysophyceae
It is close then to mix the sample (TG-MIX) fed entirely with four kinds of algaes with the sample (TG-CC-IG) of Chaetoceros.It can be seen that mixing is fed
The fatty acid sterols composition of mud blood clam children shellfish afterwards feeds chrysophyceae with single or single feeds the mud blood clam children shellfish fat of Chaetoceros
Sour sterol is close, illustrates that mud blood clam children's shellfish preference is ingested and has assimilated chrysophyceae or Chaetoceros, both algaes are institute of the present invention
The high-quality Mirco algal food of the intertidal shellfish seedling filtered out.As proof, from 4 groups of single feed microalgae 7 days after mud blood clam children shellfish
Growth it can also be seen that (table 3), single feed the sample grown speed of Chaetoceros and chrysophyceae substantially than single feed Ba Fuzao and
Nannochloropsis oculata is fast, illustrates that the result that this screening technique obtains is correct effective.
Table 3：Single feeds the size length (mm) of Mirco algal food mud blood clam children shellfish after 7 days
Claims (6)
- A kind of 1. method for screening intertidal shellfish seedling Mirco algal food, it is characterised in that described method includes the steps：1) acquisition of standard group PCA shot chartsShellfish larvae is fed as standard group by the use of mixing microalgae, feeds the aliphatic acid and sterol that shellfish larvae is determined after some cycles Composition, principal component analysis (PCA) then is carried out to aliphatic acid and sterol composition, obtains PCA shot charts；2) under same cultivating condition, feed different types of single microalgae respectively to shellfish larvae and carry out culture as experiment Group, the aliphatic acid and sterol composition of different experiments group shellfish larvae are determined after same period is fed, then to aliphatic acid and steroid Alcohol composition carries out principal component analysis, and the experiment close with standard group is filtered out according to distribution of each experimental group on PCA shot charts Bait of the microalgae of group as the shellfish.
- 2. the method as described in claim 1, it is characterised in that described shellfish is that mud blood clam, blood clam, clam, razor clam of hanging, rainbow are bright Tellin or clam.
- 3. the method as described in claim 1, it is characterised in that described microalgae is located at exponential phase of growth, and concentration reaches 80~ 120cell·μl-1。
- 4. the method as described in claim 1, it is characterised in that described shellfish is before Mirco algal food is fed, first by husky Hungry 8-10h is to empty residual bait in stomach in the natural sea-water of filter.
- 5. the method as described in claim 1, it is characterised in that described method is to use gas chromatography-mass spectrography method To carry out the quantitative analysis of aliphatic acid and sterol.
- 6. application of the method in the bait micro-algae of screening mud blood clam described in claim 1, it is characterised in that claim 1 institute The bait micro-algae of the mud blood clam for the method screening stated is Chaetoceros or chrysophyceae.
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