CN102550453A - Sea cucumber seed raising method using bacterial membrane attachment substances - Google Patents
Sea cucumber seed raising method using bacterial membrane attachment substances Download PDFInfo
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- CN102550453A CN102550453A CN2012100074326A CN201210007432A CN102550453A CN 102550453 A CN102550453 A CN 102550453A CN 2012100074326 A CN2012100074326 A CN 2012100074326A CN 201210007432 A CN201210007432 A CN 201210007432A CN 102550453 A CN102550453 A CN 102550453A
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- 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
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Abstract
The invention relates to a sea cucumber seed raising method using bacterial membrane attachment substances, which is characterized by using a transparent polyethylene corrugated plate with attached Alteromonas sp. as an attachment substrate, preferably, the Alteromonas sp. being Alteromonas sp. Lch-1 numbered as CGMCC NO.5639. By the method, attachment metamorphosis rate of young sea cucumbers can be increased by 60% as compared with that of the young sea cucumbers attached to the polyethylene corrugated plate with no bacterial membrane, cost of cultivating algal food can be reduced, and the aged polyethylene corrugated plate can be made into the bacterial membrane attachment substance to reduce the seed raising cost. The sea cucumber seed raising method has bright popularization and application prospect.
Description
Technical field
The invention belongs to sea cucumber seedling growing process field, be specifically related to a kind of sea cucumber seedling-cultivating method that utilizes the bacterial membrane adherance.
Background technology
Existing sea cucumber as cultured product has another name called imitative stichopus japonicus (Apostichopus japonicas Selenka); Belong to Echinodermata (Echinodermata); Holothuroidea (Holothuroidea) ， Parapet hand order (Aspidochirota), Stichopodidae (Stichopodidae); Imitative stichopus japonicus belongs to (Apostichopus), is typical temperate zone kind.The sea cucumber artificial breeding technique of China starts from 1954, and people such as Zhang Fengying utilize methods such as dissecting sexual gland, and auricularia has been cultivated in artificial insemination, has after this cultivated young ginseng, children's ginseng again.To the eighties in 20th century, because the establishment of factorial seedling growth new technology, the sea cucumber seedling growing process has had bigger progress.Ground such as Liaoning, Shandong particularly, the holothruian cultures industry development is swift and violent.2008, the sea cucumber nursery in Liaoning and Shandong surpassed 1000 families, emerged about 2000 tons.The Along with people's growth in the living standard, the demand of sea cucumber improves constantly.
The basic process of existing sea cucumber seedling growing process is following: 1. ovum and incubating oosperm are adopted in parent participation: the sea cucumber that the sea cucumber nursery is generally bought individual big, gonadal maturation from plant is as parent participation.Adopt nature exhaust method, method of making herbs dry gradually in the shade or stimulation by running water method to make the smart ovulation of parent participation row, control in the spawning pond smart ovum ratio at 3-5: 1 as far as possible.After fertilization carries out the branch pond with fertilized egg, and fertilized egg density is controlled at 800,000-1,000,000/m
32. larval culture: after the young hatching, select the healthy and strong young to divide the pond to cultivate, general density is 0.3-0.5/ml; The control water temperature is 20 ℃; Carry out continuous aeration, change water twice every day, each quantity of exchanged water is 30% of total breeding water body; Carry out bait throwing in according to the degree of being satiated with food of the young every day, every day three to four times.Mostly bait is bait such as salt algae, Chaetoceros muelleri, chlorella.Microtia young daily ration of feeding is 20,000 frustule/ml, and the middle ear young is 30,000 frustule/ml, and the big ear young is 40,000 frustule/ml, and adjusts daily ration, feeding quantity according to the degree of being satiated with food of the young.3. young ginseng is adhered to: the lay polyethylene film accepts the young ginseng to the bottom at the bottom of the pond when a small amount of doliolaria appears in the young, second day input adherance.Young ginseng adherance commonly used mainly contains transparent polyethylene corrugated plating, cellothene, old etting, scallop cage etc.4. young ginseng is cultivated: after young ginseng was adhered to, diatom was its optimum bait.Except that benthic diatom, also have mouse tail algae powder, artificial compound bait and ooze etc.Multiple bait can use separately, also can mix use, throws something and feeds every day twice.The control suitable temperature, continuous aeration is cultivated.Throw in adherance after 8 days, fall the pond, adopt flowing water, change method such as water and carry out the renewal of breeding water and exchange, after this remove contamination as the case may be or the work of falling the pond.When proceeding breed or releasing, can use KCl anaesthesia or direct flowing water ballistic method that young ginseng is stripped down from adherance.
The sea cucumber young adherance that the user of overwhelming majority breed now is accustomed to using is the transparent polyethylene corrugated plating, and the settlement and metamorphosis rate of the sea cucumber young on the transparent polyethylene corrugated plating is lower, is generally about 20%, has a strong impact on the development of holothruian cultures industry.Therefore, just be necessary very much to start with and improve the success rate that sea cucumber is grown seedlings from sea cucumber young adherance aspect.
Summary of the invention
The purpose of this invention is to provide a kind of sea cucumber seedling-cultivating method that utilizes the bacterial membrane adherance, to remedy the above-mentioned deficiency of existing sea cucumber seedling growing process.
Sea cucumber seedling-cultivating method of the present invention is characterized in that, is as adherance with the transparent polyethylene corrugated plating that is attached with monad Alteromonas sp. alternately.
The above-mentioned transparent polyethylene corrugated plating preparation method who adheres to monad Alteromonas sp. alternately is following:
At first oyster shells is adhered to the two sides of polyethylene corrugated plating with epoxide-resin glue, after soaking with sodium hydrate aqueous solution again, wash with the suction filtration seawater.After the flushing it is dipped in and fills in the Alteromonas sp. bacterium bacterium liquid, aeration is cultivated can obtain the bacterial membrane adherance.
Above-mentioned alternately monad Alteromonas sp.; Select the alternately monad Lch-1 (Alteromonas sp.) of screening for use; Be deposited in the China Committee for Culture Collection of Microorganisms common micro-organisms center that is positioned at No. 3, No. 1 institute in Chaoyang District Beijing North Star West Road on December 22nd, 2011, deposit number is CGMCC NO.5639.
Sea cucumber seedling-cultivating method of the present invention, its concrete steps are following: at first carry out parent participation and adopt ovum and incubating oosperm, cultivate the young with bait then; Make the bacterial membrane adherance again; Throw in the bacterial membrane adherance sea cucumber young is adhered to, the bait of throwing something and feeding is at last cultivated young ginseng, treats after most of young ginseng body reaches 15mm-20mm young ginseng to be peeled off adherance; Wherein the bait that uses is Chaetoceros muelleri the larval culture initial stage, the mixing bait of throw something and feed when growing to the middle ear young female raw cook of food and ocean rhodotorula.
The present invention can make the settlement and metamorphosis rate of the sea cucumber young use the polyethylene corrugated plating to improve 60% more separately, can reduce the cost of cultivation of algae bait again and can utilize aging polyethylene corrugated plating making bacterial membrane adherance to reduce seedling cost.
Embodiment
The present invention utilizes the sea cucumber seedling-cultivating method of bacterial membrane adherance to be divided into parent participation and adopts ovum and incubating oosperm, cultivates the young, makes the bacterial membrane adherance, adheres to young ginseng and cultivate five steps of young ginseng, makes specifying down respectively in the face of them.
1. ovum and incubating oosperm are adopted in parent participation: select sea cucumber that individual fresh weight reaches 250g, gonadal maturation as growing seedlings parent participation.Storage pond, holding pond after draining water dries in the shade parent participation, and then uses stimulation by running water, and the method that employing is dried in the shade and stimulation by running water combines can make sea cucumber lay eggs simultaneously and can improve the egg laying amount of sea cucumber.Afterwards parent participation is put into the spawning pond that water temperature has been adjusted to 20 ℃ along pool wall, constantly the hero ginseng is taken out after the ovulation of female ginseng, smart ovum ratio is at 3-5 in the control spawning pond: 1.After fertilization carries out the branch pond with siphonage, and fertilized egg density is controlled at 80 ± 50,000/m3.In the incubating oosperm process, constantly unnecessary sperm is pulled and is stirred one time per half an hour out Chi Shui, to reduce the probability of sea cucumber development of fertilized ova deformity.
2. cultivate the young: after the young hatching, the lopsided unsound young that utilizes siphonage will be sunken to bottom is drained.Select the healthy and strong young to use siphonage to carry out the branch pond; Density is 0.2-0.5/ml behind the branch pond, and the control water temperature is 18-21 ℃, carries out continuous aeration; Use every day 22 ℃ of seawater to change water twice; Quantity of exchanged water is 30% of total breeding water body, and regulate daily ration, feeding quantity according to the degree of being satiated with food of the young every day, and is full and culture no residual bait in the pond with the stomach that guarantees the sea cucumber young.Every day is bait throwing in during changing water, four times a day.Initial stage the Chaetoceros muelleri of only throwing something and feeding, feeding volume is about 20,000 frustule/ml; When growing, mix the final concentration of throwing something and feeding and be (2-4) * 10 for the female raw cook of the food of (0.1-0.5) g/m3 and final concentration to the middle ear young
4The ocean rhodotorula of cells/ml is eaten female raw cook and after grinding, is filtered with 300 mesh sieve thin,tough silk and be beneficial to the young and ingest.
3. make the bacterial membrane adherance: the oyster shells that buys is carried out 2-3 screening with 45 orders and 60 purpose mesh screens successively, obtain the oyster shells of corresponding particle diameter.Be coated onto epoxide-resin glue on the two sides of polyethylene corrugated plating respectively equably again; Form epoxy glue layer respectively; And above-mentioned oyster shells sticked to respectively on the outside of each epoxy glue layer, form the oyster shells layer respectively, be dipped in after solidifying in the sodium hydrate aqueous solution that mass percent is 1%-10%; Soaked 1-2 days, and took out the back and rinse out sodium hydroxide with filtering sea.At last; The above-mentioned polyethylene corrugated plating that scribbles epoxy glue layer and oyster shells layer is immersed in the breed pond of ZoBell 2216E culture fluid that concentration is the 10%-40% normal concentration; Volume ratio according to bacterium liquid and culture fluid is (1-4): 20 ratio adds Alteromonas sp. bacterium bacterium liquid; Aeration was cultivated 48-96 hour under water temperature 18-26 ℃ condition, formed mycoderm, and guaranteed that the bacterial density in the mycoderm surpasses 10
7Cells/cm
2, promptly obtain the bacterial membrane adherance.The Alteromonas sp. bacterium of selecting for use can be this genus bacterium commonly used, and wherein a kind of is the Alteromonas Lch-1 bacterium that screening obtains, and concrete condition is following:
Screening obtains the particle-bound bacteria that above-mentioned Alteromonas Lch-1 bacterium is gathered from Wendeng City, Shandong sea cucumber nursery through purifying.Above-mentioned its normal concentration of ZoBell 2216E culture fluid is following: peptone 5g/L; Yeast extract 1g/L; PH is 7.2.Above-mentioned Alteromonas Lch-1 bacterium bacterium liquid is the bacterium bacterium liquid of in the ZoBell of normal concentration 2216E culture fluid, cultivating 48h under 25 ℃, dark condition.Above-mentioned Alteromonas Lch-1 forms circular bacterium colony on ZoBell 2216E solid culture medium, smooth surface, and flat-ended protrusions, neat in edge, it is 1～2mm that colony diameter is about, surface wettability, white, opaque, Gram is negative.Above-mentioned Alteromonas Lch-1 has very strong attachment characteristic.
4. adhere to young ginseng: treat that the sea cucumber paedomorphosis to the big ear young later stage, drops into the bacterial membrane adherance when 5 pairs of orbicules appear in health.After five pairs of orbicules appear in big ear paedomorphosis, promptly get into the doliolaria phase in 24 hours, throw in the adhesive rate that adherance helps improving the sea cucumber young this moment.Oyster shells on the bacterial membrane adherance is beneficial to and prolongs the time of staying of bait on adherance; Be beneficial to ingesting of the sea cucumber young; The oyster shells laminar surface is attached with the bacterial membrane of the Alteromonas Lch-1 formation of high induced activity, the settlement and metamorphosis of the inducing sea cucumber young expeditiously simultaneously.After throwing in adherance, change during operations such as water, bait throwing in and the larval culture identical.
5. cultivate young ginseng: treat sea cucumber paedomorphosis to young ginseng after the stage, spirulina powder, mouse tail algae powder, aluzyme are fermented according to 10: 20: 1 ratio of quality that the fermentation back is filtered the back with 200 order mesh bags and thrown something and fed twice every day, daily ration, feeding quantity is 10g/m at every turn
3Fresh ooze is thrown something and fed with fermented composite feed after 24 hours through aeration, and daily ration, feeding quantity also is 10g/m
3Daily ration, feeding quantity is regulated according to the situation of ingesting of young ginseng.After the bait throwing in 15 minutes, stopped aeration 30 minutes.Stop aeration and help sea cucumber bait and deposit on the adherance, reduce the bait waste.Throw in adherance after 8 days, fall the pond, after this remove contamination as the case may be or the work of falling the pond.Adopt the flowing water method to carry out the renewal exchange of breeding water.The control suitable temperature carries out continuous oxygenation.When proceeding to culture or releasing, service property (quality) percentage is that 0.35% KCl sea water solution soaked after 5 minutes, is moved back in the fresh seawater, and young ginseng is separated from adherance.Adopt the KCl anaesthesia to peel off the young douche of proofreading and to reduce the frictionally damage of oyster shells young ginseng.
The present invention can make the settlement and metamorphosis rate of the sea cucumber young use the polyethylene corrugated plating to improve 60% more separately, can reduce the cost of cultivation of algae bait again and can utilize aging polyethylene corrugated plating making bacterial membrane adherance to reduce seedling cost.
Claims (4)
1. a sea cucumber seedling-cultivating method that utilizes the bacterial membrane adherance is characterized in that, is as adherance with the transparent polyethylene corrugated plating that is attached with monad Alteromonas sp. alternately.
2. seedling-cultivating method as claimed in claim 1 is characterized in that, the described transparent polyethylene corrugated plating preparation method who adheres to monad Alteromonas sp. alternately is following:
At first oyster shells is adhered to the two sides of polyethylene corrugated plating with epoxide-resin glue, after soaking with sodium hydrate aqueous solution again, wash with the suction filtration seawater; After the flushing it is dipped in and fills in the Alteromonas sp. bacterium bacterium liquid, aeration is cultivated can obtain the bacterial membrane adherance.
3. according to claim 1 or claim 2 method is characterized in that described alternately monad Alteromonas sp is monad Lch-1 alternately, and deposit number is CGMCC NO.5639.
4. the described seedling-cultivating method of claim 1, its concrete steps are following: at first carry out parent participation and adopt ovum and incubating oosperm, cultivate the young with bait then; Make the bacterial membrane adherance again; Throw in the bacterial membrane adherance sea cucumber young is adhered to, the bait of throwing something and feeding is at last cultivated young ginseng, treats after most of young ginseng body reaches 15mm-20mm young ginseng to be peeled off adherance; Wherein the bait that uses is Chaetoceros muelleri the larval culture initial stage, the mixing bait of throw something and feed when growing to the middle ear young female raw cook of food and ocean rhodotorula.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113057126A (en) * | 2021-03-19 | 2021-07-02 | 华南农业大学 | Method for cultivating standardized holothuria leucospilota seedlings |
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| US5047344A (en) * | 1983-12-30 | 1991-09-10 | Research Corporation Techniques, Inc. | Induction of settlement and metamorphosis in Crassostrea virginica by melanin-synthesizing bacteria and ammonia, and metabolic products of said bacteria |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113057126A (en) * | 2021-03-19 | 2021-07-02 | 华南农业大学 | Method for cultivating standardized holothuria leucospilota seedlings |
| CN113057126B (en) * | 2021-03-19 | 2022-06-10 | 华南农业大学 | Method for cultivating standardized holothuria leucospilota seedlings |
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Application publication date: 20120711 |