CN101099452A - A kind of artificial breeding method for intertidal zone sponge laboratory - Google Patents
A kind of artificial breeding method for intertidal zone sponge laboratory Download PDFInfo
- Publication number
- CN101099452A CN101099452A CNA2006100471255A CN200610047125A CN101099452A CN 101099452 A CN101099452 A CN 101099452A CN A2006100471255 A CNA2006100471255 A CN A2006100471255A CN 200610047125 A CN200610047125 A CN 200610047125A CN 101099452 A CN101099452 A CN 101099452A
- Authority
- CN
- China
- Prior art keywords
- sponge
- laboratory
- intertidal
- sponges
- biomass
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000009395 breeding Methods 0.000 title abstract description 4
- 238000000034 method Methods 0.000 claims abstract description 22
- 239000002028 Biomass Substances 0.000 claims abstract description 15
- 230000012010 growth Effects 0.000 claims abstract description 15
- 241000195493 Cryptophyta Species 0.000 claims abstract description 11
- 239000012535 impurity Substances 0.000 claims abstract description 6
- 238000011156 evaluation Methods 0.000 claims abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 29
- 239000013535 sea water Substances 0.000 claims description 14
- 238000011534 incubation Methods 0.000 claims description 13
- 239000011521 glass Substances 0.000 claims description 10
- 239000000463 material Substances 0.000 claims description 8
- 230000029052 metamorphosis Effects 0.000 claims description 3
- 239000003814 drug Substances 0.000 abstract description 10
- 229940079593 drug Drugs 0.000 abstract description 4
- 230000001488 breeding effect Effects 0.000 abstract description 3
- 238000012364 cultivation method Methods 0.000 abstract 1
- 238000012258 culturing Methods 0.000 abstract 1
- 241000243142 Porifera Species 0.000 description 97
- 238000002474 experimental method Methods 0.000 description 9
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 239000001301 oxygen Substances 0.000 description 7
- 229910052760 oxygen Inorganic materials 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 5
- 230000003203 everyday effect Effects 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 4
- 230000002706 hydrostatic effect Effects 0.000 description 4
- 239000004575 stone Substances 0.000 description 4
- 239000003643 water by type Substances 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 241000206751 Chrysophyceae Species 0.000 description 2
- 241000206747 Cylindrotheca closterium Species 0.000 description 2
- 241000765216 Hymeniacidon perlevis Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 230000005587 bubbling Effects 0.000 description 2
- 238000011088 calibration curve Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000009509 drug development Methods 0.000 description 2
- 244000037666 field crops Species 0.000 description 2
- 239000005338 frosted glass Substances 0.000 description 2
- 230000009605 growth rhythm Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 230000000050 nutritive effect Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 230000001932 seasonal effect Effects 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000036436 anti-hiv Effects 0.000 description 1
- 230000001857 anti-mycotic effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000002543 antimycotic Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000004568 cement Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- JEGUKCSWCFPDGT-UHFFFAOYSA-N h2o hydrate Chemical compound O.O JEGUKCSWCFPDGT-UHFFFAOYSA-N 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000011005 laboratory method Methods 0.000 description 1
- 208000020442 loss of weight Diseases 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Images
Classifications
-
- 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
- Cultivation Of Seaweed (AREA)
- Cultivation Of Plants (AREA)
Abstract
本发明涉及一种潮间带海绵在实验室条件下人工养殖获得生物量的方法，将海绵在潮间带采集后，带回实验室后首先清除海绵表面杂质及脏物，然后切割成小块，放入培养器中暂养，然后固定在载体上，放入培养器控温、避光培养，投喂饵料藻类作为主体食物，以海绵贴附状况、形态变化及生物量变化情况作为评价指标，培养10-20天可达到原代海绵生长的生物量最大值。本发明是首次在实验室中对潮间带海绵进行人工养殖，方法简单而有效，可将海洋中野生的潮间带海绵在实验室的规模下养殖生长并获得一定的海绵生物量。本发明的潮间带海绵的室内养殖方法为海绵工厂化养殖提供技术支撑，从而为解决海绵生物药物的“药源供给不足问题”提出一种新途径，具有广泛应用前景。The invention relates to a method for artificially breeding intertidal sponges to obtain biomass under laboratory conditions. After the sponges are collected in the intertidal zone and brought back to the laboratory, the impurities and dirt on the surface of the sponge are first removed, and then cut into small pieces. , put it into the incubator for temporary cultivation, then fix it on the carrier, put it into the incubator to control the temperature, and cultivate it in the dark. The bait algae is used as the main food, and the attachment status, shape change and biomass change of the sponge are used as evaluation indicators. , culturing for 10-20 days can reach the maximum biomass of primary sponge growth. The present invention is the first artificial breeding of intertidal sponges in a laboratory. The method is simple and effective, and the wild intertidal sponges in the ocean can be cultivated and grown on a laboratory scale to obtain a certain amount of sponge biomass. The indoor cultivation method of intertidal sponges of the present invention provides technical support for sponge industrial cultivation, thereby proposing a new approach for solving the "insufficient supply of drug sources" of sponge biological medicines, and has wide application prospects.
Description
Technical field
The present invention relates to the marine biotechnology field, is to adopt sponge explant cultural method to produce the technology of spongy biological amount in the laboratory, specifically set up a kind of in the laboratory method of artificial culture intertidal zone marine sponge.
Background technology
In the at present known marine organisms, sponge is the largest source of bioactive natural product, bioactive natural product as antitumor, antimycotic even anti-HIV, and other animals and plants in the high Yu Haiyang of probability of the reactive compound that discovery obtains from sponge, so the sponge drug development becomes marine drug hot of research and development in recent years.But activity substance content is few in the cavernous body, generally has only 10 of weight in wet base
-8" medicine source undersupply problem " promptly do not have enough sponges and measures the active substance that extracts some and can supply the deep research of medicine, making some medicines only rest on the clinical trial stage can't continue, thereby " medicine source undersupply problem " restrictive factor of just having become the sponge drug development obviously to lag behind.Multiple mode is attempted going to address this problem but scabrous key technology is all arranged, and sponge cell culture technology for example also not have at present the cell-line that foundation can continuous passage; Chemosynthesis costs an arm and a leg, step complexity, practical feasibility are too low; Generally believe that at present the breed of sponge industrial artificial is considered to one of mode more likely that can solve " medicine source undersupply problem ".The report of external relevant sponge breed aspect mostly relates to the sponge at deep-sea, they culture for the land for growing field crops that the breed of sponge explant concentrates in the seawater mostly, build metal truss, cement dish or set up rope at the water body of depth of water 5-10m, fix on it after sponge gathered stripping and slicing, but owing to be subjected to the influence of weather such as uncertain storm, sponge death, taken away often by storm and to take place, it is lower, uncertain relatively large to cause the land for growing field crops to culture success rate.Stone once reported under the open-air self-sow state of film sponge in great numbers with its growth area and to increase about 2 times as index between 5 months in 1970, increased average every month approximately 40%, and these 5 months also is the month that several spongy biological amounts unique in a year rise.Industrial aquaculture is because the controlled shortcoming that can avoid physioclimate to change the influence that sponge is grown of condition.Document also once had the data that is reported in laboratory aquarium breed sponge, but its objective is the growth rhythm of research variety classes and region sponge, investigated external environmental condition to the sponge growth effect, for example the temperature of Pei Yanging, salinity, ammonia nitrogen etc.The sponge of different growing environments and sharp class, to the requirement difference of growing environment, the kind difference of the active substance that is contained in the body is so the sponge artificial culture under the laboratory condition can be regulated and control according to natural environment.The present invention be directed to the cultural method of intertidal zone marine sponge, under experiment indoors artificial controlled condition, cultivate intertidal zone marine sponge and obtain biomass for the researcher use of doing its related fields research.
Summary of the invention
It is effective and simple to operate to the purpose of this invention is to provide a cover, thereby can carry out the method that the spongy biological amount is produced in indoor controlled cultivation to intertidal zone marine sponge.
The method principle: intertidal zone marine sponge is a lower animal of seeking the non-selectivity filter food of growing admittedly, and regeneration capacity is stronger, under suitable external environment, is attached at continued growth on other carriers but cut an explant from sponge.Its influence that is subjected to seasonal temperature of growing of film sponge in great numbers that is grown in the temperate zone is bigger, presents that spring and summer seasonal growth is vigorous, the growth rhythm of autumn and winter season decline, cultivates for the indoor control of sponge and transfers part that theoretical foundation is provided.
For achieving the above object, the technical solution used in the present invention is:
A kind of intertidal zone marine sponge laboratory artificial cultivating method that is used for, laboratory are manually made lower animal-sponge that the suitable growth environment is cultivated intertidal zone set growth; Sponge self-sow environment with manual simulation intertidal zone, laboratory set growth is produced the spongy biological amount; Concrete steps, after the sponge collection that is grown on the reef of intertidal zone, put into immediately in the keg of containing seawater, take back the laboratory and remove sponge surface impurity and foul, then sponge is cut into the fritter of suitable size, putting into glass jar supports temporarily, be fixed on the carrier then, place in the incubator, temperature control, lucifuge are cultivated, throw something and feed feed algae as main body food, attach situation, metamorphosis and biomass situation of change as evaluation index, cultivate the maximum biomass that can reach former generation sponge growth in 10-20 days with sponge.The present invention has overcome the sponge laboratory and has cultured growth phase to problem slow, that the cycle is long.
Specific operation process is:
When 1) sponging on the reef of intertidal zone, seashore, select bright-colored, surperficial relatively flat, projection is short and sturdy, surface clean (the impurity foul is less) sponge places sponge after the collection in the keg of containing seawater immediately;
2) after sponge is taken back the laboratory, when carefully clearing up sponge body surface impurity foul, have in the spongy tissue than high concentration of sediment stone grain, then abandon selecting other sponges,, then carefully take out with tweezers if the stone grain is less and relatively large in the tissue if run into tweezers and injection needle;
3) aseptic operation cutter cutting sponge, suitable size is the about 3-9cm of area
2, to put into glass jar and supported 2-7 days temporarily, the wound that forms during to its cutting and collection tentatively heals, and finishes preliminary callus process, and temperature adapted when gathered on foster temporarily temperature and seashore, and then was adjusted to cultivation temperature gradually;
4) sponge is fixed on to place in the reactor on the carrier with fine rule and cultivates, and the carrier size is about 4-8 times of sponge size;
5) control sponge culture density 2-15 ‰ (m/v), water temperature 15-25 ℃, oxyty 5-9mg/L;
6) sunlight direct projection sponge is avoided in the place that hides one deck brown paper or directly reactor is placed on no sunlight direct projection, glass jar outside;
7) in the culture period, keep cultivating the NH in the water body
4 +Concentration is lower than 1mg/L;
8) concentration of throwing something and feeding is 2 * 10
4-2 * 10
5The unit cell algae as food;
Every day is changed water in quiet cultivation, and sponge is a main body food with the feed algae of throwing something and feeding etc.;
It is the algae concentration of throwing something and feeding according to streamflow calculating every day incubator planted agent that stream adds training method;
8) in the incubation, sponge attaches and sprawls, metamorphosis is taken pictures and record with digital camera, and biomass is weighed with weight in wet base (wet sponge quality), is converted into the weight in wet base increased percentage after the weighing, and formula is as follows: S=(M
n-M
0)/M
0* 100%, M
0, M
tBe respectively sponge and cultivating the weight in wet base of the 0th day and n days;
9) look the different cultivations of sponge collection season state 10-20 days, can reach the maximum biomass of former generation sponge growth, winter-spring season was cultivated 15-20 days, and summer and autumn cultivated 10-15 days.
The present invention compared with prior art has following advantage:
1. technology is complete relatively, first film sponge in great numbers is provided a cover complete method and is used for laboratory artificial cultivating.
2. simple to operation, sponge carrier making, fixing means, temperature control, illumination control, dissolved oxygen concentration control etc. are all very simple, reach requirement easily.
3, be widely used, the method can be used for producing in a short time film sponge biomass of former generation in great numbers; Can provide sponge to study the excellent material of international difficult point-cell culture by the good sponge of training quality; Can carry out the go down to posterity comparison of culture studies and targeted activity material regulation and control of spongy tissue piece.
In a word, the present invention can provide film sponge in great numbers effectively to cultivate under the controlled condition of laboratory and produce sponge biomass of former generation, technology is more complete relatively, easy and simple to handle, produce the spongy biological amount in indoor controlled cultivation, for the sponge industrial aquaculture provides technical support, thereby, has wide application prospect for " the medicine source undersupply problem " that solves the spongy biological medicine proposes a kind of new way.
Description of drawings
Fig. 1 is 5L water purification cultivating system figure; Wherein: 1 sponge is cultivated glass jar, and 2 sponges attach carrier and orange sponge, 3 breather pipes, 4 gas stones, 5 air pumps; 6 waters cylinders, 7 temperature controllers, 8 desks, 9 waters water storage tanks, 10 shallow water pumps, 11 water-bath pipes;
Fig. 2 is 5L flowing water cultivating system figure; Wherein: 12 air, 13 ozone generators, 14 stirring arms, 15 bait algae liquid, 16 mixing tanks, 17 air filter membranes, 18 sponge incubators, 19 waste collection grooves, 20 heat exchangers, 21 seawater, 22 Seawater Treatment grooves, 23 remove the albumen device;
Fig. 3 is outward appearance growth conditions figure in the film sponge incubation in great numbers; Wherein: a is for cultivating initial sponge state; Sponge state when b finishes for cultivating;
Fig. 4 is film sponge in great numbers biomass change curve in the hydrostatic incubation;
Fig. 5 adds biomass change curve in the incubation for film sponge in great numbers at stream;
Fig. 6 is the GC-MS spectrogram of 5 α in the sponge extract-Cholestan-3 beta-ol;
Fig. 7 is 5 α-Cholestan-3 beta-ol calibration curve.
Embodiment
Below by specific embodiment method of the present invention and result are described, the present invention uses and buys deep-sea water as breeding seawater, is used for the cultivation of sponge breed and feed algae; Weight in wet base adopts analytical balance to take by weighing quality, gets the sponge growth quality by the loss of weight method, and mercurial thermometer is measured the interior actual water temperature of cultivation water body and is convenient to regulate; Portable dissolved oxygen meter is measured the air mass flow that dissolved oxygen concentration is convenient to regulate the air pump supply in the water body, and the cultivation of bait algae adopts filtration sterilization seawater conical flask to cultivate, and nutritive salt adopts the general Kang Wei mill nutritive salt of algae culture.
5L hydrostatic system is cultivated.
Intertidal zone film sponge in great numbers (Hymeniacidon perleve) is collected in DaLian, China Ling Shui bridge intertidal zone in July, 2005, and sponge surface impurity and foul are less, choose the about 2 * 2cm of area during collection
2Fritter, take back the laboratory and be placed in the 12L glass jar of containing seawater and supported 2 days temporarily, support temperature temporarily and be controlled at about 20 ± 1 ℃.Cultivating system as shown in Figure 1.Cultivate initial, be fixed on sponge on the frosted glass carrier of 4 * 4cm with line, put into the 5L glass jar that fills seawater for totally 15, then glass jar is placed temperature to be about in 20 ± 2 ℃ the waters, air pump and gas distributor bubbling air, dissolved oxygen meter is surveyed dissolved oxygen in the water body, the size of regulating the bubbling air flow makes dissolved oxygen concentration in the water in 7 ± 0.5mg/L scope, the glass jar outside covers with brown paper and prevents that sunlight from shining directly on the cavernous body, change water 100% every day, change and will lay in seawater before the water earlier and be adjusted to sponge cultivate identical temperature in the waters, change throw something and feed behind the water Nitzschia closterium minutissima and the equilateral chrysophyceae of ball as food, concentration respectively is 2 * 10
5/ ml.In the culture period,, cultivate the sponge weight in wet base and be in plateau relatively and stop experiment every the weight in wet base of 2 days weighing sponges and carrier.With 5 α-Cholestan-3 beta-ol as the targeted activity material in the film cavernous body in great numbers, after cultivating end, the content of targeted activity material when testing initial and end with GC-MS logotype method detection cavernous body is inherent, the GC-MS spectrogram of 5 α in the sponge extract-Cholestan-3 beta-ol as shown in Figure 6, according to 5 α-Cholestan-3 beta-ol content in peak area and calibration curve (Fig. 7) calculation sample.The result, in the incubation, it is more bright-coloured when initial that the sponge color becomes, sponge has bigger attaching and sprawls on carrier, change has taken place in form, has become from original smooth surface to have a lot of full long projection (sprout) shapes (as shown in Figure 3), and this sprout endogenous cells content is abundant than other positions of sponge, be the good material of sponge cell culture, the NH in the incubation in the water body
4 +Concentration is lower than 1mg/ (as shown in table 1) all the time, tests end in the 11st day, and survival rate reaches 100%, and the spongy biological amount rises to 140% (as shown in Figure 4) of original weight in wet base, increases average every day to surpass 4%.5 α-Cholestan-3 beta-ol content in the cavernous body has also risen to 10.478mg/g dry weight sponge (as table 3) by the 6.244mg/g dry weight sponge before the experiment, is about experiment 1.5 times when initial.
Table 1 is cultivated the interior NH of different incubation time water bodys down for hydrostatic
4 +The concentration table;
| NH4(mg/L) | 0 | 1 | 3 | 5 | 7 | 9 |
| |
0.045 | 0.606 | 1.159 | 0.855 | 0.872 | 0.648 |
| |
0.045 | 0.655 | 1.174 | 0.815 | 0.741 | 0.621 |
Table 2 is cultivated the interior NH of different incubation time water bodys down for flowing water
4 +The concentration table;
| Incubation time (my god) | 4 | 6 | 8 | 10 | 12 | 14 | 16 |
| Ammonia nitrogen concentration (mg/L) in the flowing water water body | 0.264 | 0.587 | 0.511 | 0.313 | 0.293 | 0.398 | 0.382 |
Table 3 is cultivated interior 5 α of cavernous body-Cholestan-3 beta-ol content table down for hydrostatic;
| Experimental point | Test initial (0day) | Experiment finishes (11day) |
| Peak area | 5410 | 11033.5 |
| 5 α-Cholestan-3 beta-ol content (mg/g dry weight sponge) | 6.244 | 10.278 |
5L stream adds system and cultivates.Film sponge in great numbers (Hymeniacidon perleve) is collected in DaLian, China Ling Shui bridge intertidal zone in July, 2005, takes back the laboratory and is placed on temporary supporting 2 days in the 12L glass jar of containing seawater, and temperature is controlled at 18-20 ℃.Cultivating system as shown in Figure 2.Cultivate initial, be fixed on sponge on the circular frosted glass carrier of the about 5cm of diameter with line, put into the 5L liquid that fills seawater and rise incubator for totally 15, discharge is 20L/day, the control of incubator coated outside cooling tube is cultivated water temperature at 20 ± 1 ℃, and air pump and gas distributor provide sponge required oxygen, and whole system places indoor no sunlit place, throw something and feed Nitzschia closterium minutissima, the equilateral chrysophyceae of ball as food, and concentration respectively is 2 * 10
5/ ml, the NH in the water body is cultivated in control in the incubation
4 +Concentration is no more than 1mg/L.In the culture period, measured the spongy biological amount every 2 days, experiment appears stopping when sorrow is moved back phenomenon in the sponge apparent condition.With 5 α-Cholestan-3 beta-ol as the targeted activity material in the film cavernous body in great numbers, the content of targeted activity material when initial and end are tested with the method detection cavernous body inherence of GC-MS logotype in cultivation end back, the result, in the incubation, it is more bright-coloured when initial that the sponge color becomes, and sponge has bigger attaching on carrier sprawls, and change has taken place form, become from original smooth surface and to have had a lot of full long overshooting shapes, the interior NH of water body in the incubation
4 +Concentration is lower than 0.6mg/L (as shown in table 2) all the time, tests end in the 16th day, and the sponge survival rate is 87%.The spongy biological amount rises to 140% (as shown in Figure 5) of original weight in wet base, the increment of average every day has all surpassed 4%, 5 α-Cholestan-3 beta-ol content in the cavernous body has also risen to 10.733mg/g dry weight sponge (as table 4) by the 6.721mg/g dry weight sponge before the experiment, is about experiment 1.5 times when initial.
Table 4 is cultivated interior 5 α of cavernous body-Cholestan-3 beta-ol content volumeization table down for flowing water;
| Experimental point | Test initial (0day) | Experiment finishes (11day) |
| Peak area | 7499.5 | 16412.3 |
| 5 α-Cholestan-3 beta-ol content (mg/g dry weight sponge) | 6.721 | 10.733 |
Claims (5)
1. one kind is used for the intertidal zone marine sponge laboratory artificial cultivating method, and lower animal-sponge that the suitable growth environment is cultivated intertidal zone set growth is manually made in the laboratory; It is characterized in that: after sponge is gathered in the intertidal zone, at first remove sponge surface impurity and foul after taking back the laboratory, cut into fritter then, putting into incubator supports temporarily, be fixed on then on the carrier, put into incubator temperature control, lucifuge cultivation, the feed algae of throwing something and feeding is as main body food, attach situation, metamorphosis and biomass situation of change as evaluation index with sponge, cultivate the maximum biomass that can reach former generation sponge growth in 10-20 days.
2. in accordance with the method for claim 1, it is characterized in that: when sponging on the reef of intertidal zone, select bright-coloredly, surperficial relatively flat, projection are short and sturdy, table and clean sponge places sponge in the keg of containing seawater standby immediately after the collection.
3. in accordance with the method for claim 1, it is characterized in that: adopt aseptic operation cutter cutting sponge, suitable area size is about 3-9cm
2The carrier size is 4-8 a times of sponge area; Sponge carrier can be selected glass or plastic material, and sponge selects fine rule to fix.
4. in accordance with the method for claim 1, it is characterized in that: sponge will be supported 2-7 days after taking back the laboratory temporarily, and the wound that forms during to its cutting and collection tentatively heals, and temperature adapts when supporting the collection of temperature and seashore temporarily, and then be adjusted to cultivation temperature, 15-25 ℃ of sponge cultivation temperature gradually.
5. in accordance with the method for claim 1, it is characterized in that: sponge culture density 2-15 ‰ (m/v), cultivate water body oxyty 5-9mg/L, keep cultivating the NH in the water body
4 +Concentration is lower than 1mg/L; Concentration 2 * 10 when throwing something and feeding feed algae food
4-2 * 10
5Cells/ml; Avoid sunlight direct projection sponge in the incubation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2006100471255A CN101099452B (en) | 2006-07-05 | 2006-07-05 | A kind of artificial breeding method for intertidal zone sponge laboratory |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2006100471255A CN101099452B (en) | 2006-07-05 | 2006-07-05 | A kind of artificial breeding method for intertidal zone sponge laboratory |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101099452A true CN101099452A (en) | 2008-01-09 |
| CN101099452B CN101099452B (en) | 2010-07-21 |
Family
ID=39034057
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2006100471255A Expired - Fee Related CN101099452B (en) | 2006-07-05 | 2006-07-05 | A kind of artificial breeding method for intertidal zone sponge laboratory |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101099452B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102721434A (en) * | 2012-06-19 | 2012-10-10 | 哈尔滨理工大学 | Intelligent control testing system for researching intertidal benthos based on wireless network |
| CN102071166B (en) * | 2009-11-20 | 2012-12-26 | 中国科学院大连化学物理研究所 | Method for tissue artificial culture of hymeniacidon perleve by using program for temperature regulation and control |
| CN102901800A (en) * | 2012-10-24 | 2013-01-30 | 厦门大学 | Automatic simulation device for intertidal zone environmental change |
| CN105010186A (en) * | 2015-07-15 | 2015-11-04 | 厦门大学 | Method for directionally inducing attachment and metamorphosis of sponge larva |
| CN105123561A (en) * | 2015-07-15 | 2015-12-09 | 厦门大学 | Method for cultivating spongia |
| CN111670846A (en) * | 2020-04-20 | 2020-09-18 | 厦门大学 | A kind of method of spongy juvenile in situ cultivation |
-
2006
- 2006-07-05 CN CN2006100471255A patent/CN101099452B/en not_active Expired - Fee Related
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102071166B (en) * | 2009-11-20 | 2012-12-26 | 中国科学院大连化学物理研究所 | Method for tissue artificial culture of hymeniacidon perleve by using program for temperature regulation and control |
| CN102721434A (en) * | 2012-06-19 | 2012-10-10 | 哈尔滨理工大学 | Intelligent control testing system for researching intertidal benthos based on wireless network |
| CN102901800A (en) * | 2012-10-24 | 2013-01-30 | 厦门大学 | Automatic simulation device for intertidal zone environmental change |
| CN102901800B (en) * | 2012-10-24 | 2014-07-09 | 厦门大学 | Automatic simulation device for intertidal zone environmental change |
| CN105010186A (en) * | 2015-07-15 | 2015-11-04 | 厦门大学 | Method for directionally inducing attachment and metamorphosis of sponge larva |
| CN105123561A (en) * | 2015-07-15 | 2015-12-09 | 厦门大学 | Method for cultivating spongia |
| CN105123561B (en) * | 2015-07-15 | 2018-12-04 | 厦门大学 | The method of spongia transplanting |
| CN111670846A (en) * | 2020-04-20 | 2020-09-18 | 厦门大学 | A kind of method of spongy juvenile in situ cultivation |
| CN111670846B (en) * | 2020-04-20 | 2021-10-15 | 厦门大学 | A kind of method of spongy juvenile in situ cultivation |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101099452B (en) | 2010-07-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104430079B (en) | Vegetable, Oryza sativa L., flower planting support, with net cage set, the apparatus and method combined | |
| CN101099452B (en) | A kind of artificial breeding method for intertidal zone sponge laboratory | |
| CN102356746A (en) | Technology for cultivating organic fruit and vegetable matrix | |
| CN111248139B (en) | Culture device and method for high-density rotifers | |
| CN102668831A (en) | Submerged plant breeding blanket and application thereof | |
| CN104126494A (en) | Zostera marina indoor long-term cultivation method and device | |
| CN101306880A (en) | Method for controlling eutrophic water body by intercropping plants on water | |
| CN106577409A (en) | Method for inducing copepoda to produce diapause eggs | |
| CN102037891B (en) | Chinese cymbidium pot culture substrate | |
| CN101381164A (en) | Method for planting green fodder and purifying water quality based on pond-type biological floating raft | |
| CN105580763A (en) | Artificial low-salt fry breeding method for eleutheronema tetradactylum | |
| US20180263200A1 (en) | A system for cultivating aquatic plants and method thereof | |
| CN102138542A (en) | Method for reserving, conserving and culturing daphnia magna | |
| CN103004576A (en) | Method for artificially cultivating Sargassum thunbergii in intertidal zones | |
| CN102657071A (en) | Method for culturing phytoplankton with predominant chlorophyta | |
| CN105191845B (en) | A kind of method of sea cucumber and the mixed breeding of hippocampus stereo ecological | |
| CN106857226A (en) | A kind of method of Sargassum horneri in northern productivity artificial breeding | |
| CN201160430Y (en) | Cultivation device for intermediate cultivation of blue crab seedlings | |
| CN114532264B (en) | Staged pond culture method for improving pearl quality of pinctada maxima | |
| CN101322477B (en) | Method for artificially collecting and cultivating larva of intertidal belt sponge with embryo | |
| CN101692787B (en) | Method for collecting and culturing ulva spores | |
| CN109169177A (en) | A kind of environment protection flower pot and its medium production method with fermented cow dung matrix | |
| CN116098109A (en) | A pond industrialized farming system | |
| CN104872027B (en) | Facility and method for indoor large-scale cultivation of larvae and juveniles of Sillago sihama using prawn cultivation wastewater | |
| CN103663699A (en) | Method for controlling eutrophic water body by selecting grass type lake native aquatic plant |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20100721 Termination date: 20130705 |