CN102017908B - Water-saving sea cucumber culturing method - Google Patents
Water-saving sea cucumber culturing method Download PDFInfo
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- CN102017908B CN102017908B CN2010102710098A CN201010271009A CN102017908B CN 102017908 B CN102017908 B CN 102017908B CN 2010102710098 A CN2010102710098 A CN 2010102710098A CN 201010271009 A CN201010271009 A CN 201010271009A CN 102017908 B CN102017908 B CN 102017908B
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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 discloses a water-saving sea cucumber culturing method, which comprises: selecting a culturing water source; treating pool water; treating feed; controlling a water change period; and controlling a culturing environment by using a bioecological method according to culturing water ecological principles, growth environment requirements of marine organisms, correlations among organisms and harmless culturing standards. The key technique is to add bdellovibrio bacteriovorus nectophore bacterial solution into water, soak the feed in bdellovibrio bacteriovorus nectophore bacterial solution and change the water in the pool every 40 days in a culture period. The method is a biological culturing method, is green, safe, environmentally-friendly and obvious in effect, can improve water quality, promote the ingestion of sea cucumbers, improve immunity, promote growth and improve survival rate and has a bright development prospect. The method has a great significance for improving economic benefits, food safety and protecting human health.
Description
Technical field
The invention belongs to holothurian cultural technique field, particularly a kind of raising sea cucumber survival rate and immunity promote that sea cucumber grows, improves water quality fast, saves water and energy, the nuisanceless breed new method of sea cucumber of green low-carbon.
Background technology
Sea cucumber, formal name used at school are imitated stichopus japonicus (Apostichopus japonicus), have very high edibility, rank among " eight delicacies ".China's year consumption dried sea-cucumber reaches 5000 tons approximately, and supply demand gap is huge.For this reason, China has started the upsurge of apostichopus japonicus culture in the nineties.
Along with the cumulative year after year of holothruian cultures, cultivation density is excessive, the deterioration of carrying capacity overload and breeding environment, and the sea cucumber disease is also in frequent outburst, the putrid skin disease of mature stage commonly, swollen mouth disease etc.At present, the sea cucumber prevention and treatment of diseases mainly leans on the method for antibiotic dipping to handle.The long-term antibiotic that uses not only makes the pathogen pesticide resistance increase, and disease more frequently takes place, and the excessive antibiotic and the use of chemicals also cause the antibiotic residue in the sea cucumber body to exceed standard, and influence the marketing quality of sea cucumber, and the safe in utilization of people threatened.
The use severe contamination water quality of other type chemical classes disinfectant is destroyed environment.Though Chinese herbal medicine can have drug residue free, advantage that economic benefit is high, it is slow to take effect, and can't resolve epidemic disease in a short time.
On the other hand, at present the aquaculture model of sea cucumber needs a large amount of water and frequent ponds of falling changed, and has not only wasted human and material resources; The polluted water environment; And a large amount of waters causes the littoral zone seawater invasion easily, causes the soil salinization of soil, caused harm to ecotope.Method according to commonly used is handled waste water, recycles, and then needs higher investment, has increased aquaculture cost.
Thereby seek survival rate and the immunity that a kind of more efficient, more economical way improves sea cucumber, seem very urgent." green cultivation " not only overcome the side effect that the antibiotic abuse brings, and can also improve water quality, strengthens seed immunity, improves survival rate and the quality of sea cucumber, promotes the sea cucumber growth, becomes the great demand of country in recent years.The mode of at present, carrying out holothruian cultures with the Bdellovibrio telotroch is not at home and abroad appeared in the newspapers as yet.
Existing research shows, as a kind of effective microorganism preparation, no matter is in food industry or in fields such as (ocean) aquacultures, the application of Bdellovibrio all is safe.For example: abroad; Lenz and Hespell (1978) discovers; Bdellovibrio and animal and people's cell do not had an infectivity [Lenz R.W.; Hespell R.B.Attempts to grow bedellovibrios micurgically-injected into animal cells.Archives of Microbiology, 1978,119 (3): 245-248].At home, Lin Mao etc. (2006) have studied the effect of Bdellovibrio to fish cell, find that it does not have dissemination [Lin Mao, Yang Xianle, Xue Hui, Cao Haipeng, Qiu Junqiang to the fish bacterium.The effect of 02 pair of fish cell of Bdellovibrio BDH21 and pathogen.The microbiology circular, 2006,33 (1): 7-11].
Summary of the invention
For overcoming the deficiency that exists in the above-mentioned existing cultural technique, the object of the present invention is to provide a kind of water-saving holothurian culture method; This method realizes through in feed and water body, adding Bdellovibrio telotroch bacterium liquid, specifically comprises the breed choosing source, the processing of Chi Shui, the processing of feed; The control of exchange water cycle, its characteristic is learned principle according to breeding ecological, halobiontic growing environment requirement, correlation between the biology and nuisanceless breed standard; The artificial breeding environment of creating reduces input cost, improves water quality; Improve sea cucumber immunity, promote that sea cucumber grows fast, increase economic efficiency.
The object of the invention is realized through following proposal: a kind of water-saving holothurian culture method comprises following operating procedure:
(1) gets the free of contamination seawater in marine site as breeding water; Breeding water injects and cultures the pond through sand filtration, post precipitation; Culture the corrugated plating of placing in the pond through sterilization (apposition growth that is used for sea cucumber);
(2) in culturing the pond, add Bdellovibrio telotroch bacterium liquid, and strengthen aeration, stir, make the concentration of the Bdellovibrio telotroch of Chi Shuizhong reach 10～10
7Pfu/mL;
(3) in culturing the pond, throw in the sea cucumber seedling;
(4) culture during normal aeration, the control water temperature is at 15～30 ℃, the salinity of Chi Shui remains on 23 ‰～30 ‰, daily ration of feeding be the sea cucumber body weight mass percent 3～5%, bait is before input, using concentration is 10～10
7The Bdellovibrio telotroch bacterium liquid of pfu/mL soaks 15～30min; Whenever adding Bdellovibrio telotroch bacterium liquid after changing Chi Shui (full pond) once, at every turn to change (full pond) water in 5～40 days makes the Bdellovibrio telotroch bacterial concentration of Chi Shuizhong reach 10～10
7Pfu/mL.
The said breed of step (1) pond cleaning and sterilizing before injecting breeding water is clean; 3m in the said breed pond
2～4m
2Arrange 1 gas exhausting stone (being used for inflation).
The method of the said adding in step (2) and (4) Bdellovibrio telotroch bacterium liquid is: behind the Chi Shui dilution bacterium liquid of culturing in the pond, and the full pond of evenly splashing.
The aeration rate of the said increasing aeration of step (2) is 90～100L/h/m
3The aeration rate of the said normal aeration of step (4) is 70～80L/h/m
3
The said input of step (3) sea cucumber seedling when 15～30 ℃ of water temperatures according to following standard: when sea cucumber seedling specification was 400/kg, throwing in density was 600/m
2When sea cucumber seedling specification was 200/kg, input density was 400 tails/m
2When sea cucumber seedling specification was 100/kg, input density was 300 tails/m
2
The said bait of step (4) is mixed bait, and every day, bait throwing in was twice; Whenever adopted the soil pick-up at the bottom of siphonage is cultured the pond at a distance from 5～7 days.
Step (2) and (4) said Bdellovibrio are Bdellovibrio (Bdellovibrio sp.Strain) BDM01; Said Bdellovibrio is preserved in the Chinese typical culture collection center in the Wuhan University of Luojia Mountain, Wuhan, Hubei Province on April 28th, 2008, and deposit number is CCTCC NO:M 208066.BDM01 carries out ultraviolet mutagenesis by Bdellovibrio BDJ01 (deposit number is CCTCC M 208011) and obtains; It is carried out morphologic observation under electron microscope after the negative staining: Bdellovibrio BDM01 is unicellular, arc, and size is 1.7 * 1.0 μ m; End is given birth to flagellum, and flagellum length is 3.5 μ m; It is cultivated the transparent circular plaque that can form diameter 3-4mm in three days with the double-layer plate method in 28 ℃.
Said Bdellovibrio leech plastid bacterium liquid prepares according to following method: inoculation 0.5mL concentration is 10
7Cfu/mL Aeromonas hydrophila (Aeromonas hydrophila; Purchase DSMZ of Institute of Micro-biology, numbering GIM1.172 in Guangdong Province) in the nutrient broth liquid nutrient medium, cultivate 20h for 26～30 ℃; Culture fluid is behind the centrifugal 10～20min of 5000～8000rpm; Deposition is added in the DNB liquid nutrient medium after suspending with 1mL physiological saline, inserts 1mL again and contains 10
3The bacterium liquid of pfu/mL Bdellovibrio BDM01; Cultivate 36～48h in 28 ℃; Culture fluid is following behind the centrifugal 15～20min of 6000～8000rpm at 4 ℃; The supernatant that will contain the Bdellovibrio telotroch is behind the centrifugal 15～20min of 16000～18000rpm, and deposition use DNB liquid nutrient medium, water, physiological saline or 0.2mol/L pH value are 7.2～7.6 phosphate buffer suspension, and obtaining concentration is 10
9The Bdellovibrio telotroch bacterium liquid of cfu/mL.
Said DNB liquid nutrient medium is that nutrient broth 0.8g, caseinic acid hydrolysate 0.5g and yeast extract 0.1g are dissolved in the 1000ml distilled water, regulates pH value to 7.2～7.6.
Compared with prior art, the present invention has following beneficial effect:
Developing nuisanceless breed, improve the immunity of sea cucumber, promote the quick growth of sea cucumber, reduce production costs, increase economic efficiency, is the only way of China's mariculture industry development; Bdellovibrio can be improved the sea cucumber gastrointestinal tract environment, improves immunity, promotes the sea cucumber growth, and to the holothurian free of toxic effects, to human no pathogenicity.
Can be divided into the leech plastid state that breaks away from host bacteria, has the telotroch state of flagellum, free swimming and in host bacteria, grow the history of life of Bdellovibrio.When the Bdellovibrio telotroch is invaded the host, on the host cell wall, produce a hole, invade host's periplasmic space when Bdellovibrio after hole seal again.Telotroch loses flagellum and host cell is called as " leech plastid " together.Next, the hydrolysis of host's periplasmic space, the Bdellovibrio cell is elongated, and splits into a plurality of short segments, grows flagellum, spins off from host cell through discharging the dissolubility enzyme, forms telotroch again.
It is very fast that the telotroch Bdellovibrio is invaded the process of host bacteria, can accomplish in general several seconds.For Bdellovibrio leech plastid, though exist the weakness that preparation is loaded down with trivial details, be difficult for to preserve, with the control of telotroch Bdellovibrio and eliminate pathogenic or potentially pathogenic organism rapid-action, energetic.Soak feed with the telotroch Bdellovibrio, and the certain density telotroch Bdellovibrio bacterium liquid of splashing in the pool water within, the growth of sea cucumber can be promoted; Improve immunity, can control pathogenic bacteria again, purify water; And length can be changed (full pond) water in 40 days once, thereby had overcome side effect and a large amount of breeding wastewater exhaust emission environment that the antibiotic abuse brings, used manpower and material resources sparingly; For the mankind provide safety, green, non-harmful aquatic products, improve raiser's income.
Embodiment
Below in conjunction with embodiment the present invention is done further detailed description, but embodiment of the present invention is not limited thereto.
Practical implementation of the present invention place is holothruian cultures field, Shandong, and the sea cucumber kind is imitative stichopus japonicus (Apostichopus japonicus).The enforcement time is 4 months.Experimental field the source quality in marine site meets State Standard of the People's Republic of China's water quality standard for fishery (GB11607-89) requirement; Get into after the sand filtration of seawater process, the precipitation process and culture the pond, meet (NY5052-2001) requirement of Ministry of Agriculture's " pollution-free food mariculture water water quality ".The breed pond of sea cucumber is built in and is cultured in the booth, selects physical health, individual children's ginseng of uniform size, and the long 3～5cm of average body adopts the randomized block experiment design, according to 60～80/m
2Cultivation density drop in the pond.Be equally divided into 4 experimental group of A-D, B, C, each experimental group of D have 8 groups, are respectively B1, C1, D1; B2, C2, D2; , B3, C3, D3; B4, C4, D4; B5, C5, D5; B6, C6, D6; B7, C7, D7; B8, C8, B8; The every group of B1-D8 is further divided into 4 groups, i.e. B1-1～B1-4 ... D8-1～D8-4, every group of 2 repetitions.Test processing is following:
A group: normal flowing water, the manufactured feed feed of throwing something and feeding (breeding way of plant produced)
B group: the Bdellovibrio telotroch bacterium liquid of in water, splashing makes that Bdellovibrio telotroch concentration reaches 10,10 respectively in the water body
3, 10
5, 10
7Pfu/mL, the manufactured feed of throwing something and feeding;
The C group: the concentration of throwing something and feeding is 10,10
3, 10
5, 10
7Manufactured feed after the Bdellovibrio telotroch bacterium liquid of pfu/mL soaks;
D group: in water, add Bdellovibrio telotroch bacterium liquid, make cell concentration reach 10,10 respectively
3, 10
5, 10
7Pfu/mL, the concentration of throwing something and feeding respectively simultaneously is 10,10
3, 10
5, 10
7Manufactured feed after the Bdellovibrio telotroch bacterium liquid of pfu/mL soaks;
B1, C1, D1 organize per 5 days and change (full pond) water once, add bacterium again after changing water;
B2, C2, D2 organize per 10 days and change (full pond) water once, add bacterium again after changing water;
B3, C3, D3 organize per 15 days and change (full pond) water once, add bacterium again after changing water;
B4, C4, D4 organize per 20 days and change (full pond) water once, add bacterium again after changing water;
B5, C5, D5 organize per 25 days and change (full pond) water once, add bacterium again after changing water;
B6, C6, D6 organize per 30 days and change (full pond) water once, add bacterium again after changing water;
B7, C7, D7 organize per 35 days and change (full pond) water once, add bacterium again after changing water;
B8, C8, D8 organize per 40 days and change (full pond) water once, add bacterium again after changing water;
B1-1, C1-1, D1-1 ... The bacterial concentration that D8-1 throws in is 10pfu/mL;
B 1-2, C1-2, D1-2 ... The bacterial concentration that D8-2 throws in is 10
3Pfu/mL;
B1-3, C1-3, D1-3 ... The bacterial concentration that D8-3 throws in is 10
5Pfu/mL;
B1-4, C1-4, D1-4 ... The bacterial concentration that D8-4 throws in is 10
7Pfu/mL;
Feeds utilized People's Republic of China's agricultural industry criteria " the manufactured feed safety limit is used in the pollution-free food fishing " regulation (NY5072-2002) that meets.Duration of test is thrown something and fed 2 every day, and morning, evening are respectively once.Growth indexes, immune indexes and the microorganism situation of sea cucumber measured in off-test.
1, the assay method of sea cucumber growth indexes is following:
Feed after 4 months, add up the sea cucumber number of surviving in final every mouthful of pond, randomly draw 100 sea cucumbers respectively from every mouthful of pond, with its body of vernier caliper measurement long (mm), electronic balance is surveyed its weight (g), calculates following parameter:
(1) survival rate (%)=N
t/ N
o* 100%;
(2) rate of body weight gain (%)=(W
t-W
o) W
o* 100%;
(3) the long growth rate (%) of body=(L
t-L
o)/L
o* 100%;
N in the formula
oTotal number of sea cucumber during for on-test; N
tTotal number of sea cucumber during for off-test; W
oSea cucumber body weight during for on-test; W
tSea cucumber body weight during for off-test; L
oThe sea cucumber body is long during for on-test; L
tThe sea cucumber body is long during for off-test;
2, the mensuration of the immune indexes of sea cucumber
Alkaline phosphatase (AKP) and acid phosphatase (ACP) activity all adopt Kruzel (1982) disodium phenyl phosphate method,
Blood is extracted in 1/3 puncture behind the outside of belly mouth of sea cucumber, places centrifuge tube, and in 4 ℃, the centrifugal 10min of 3000r/min gets supernatant, is the serum of sea cucumber.Place ice chest to preserve.
The mensuration of acid phosphatase (ACP) adopts the disodium phenyl phosphate method., produce 1mg phenol person and be defined as an enzyme activity unit at 37 ℃ and substrate disodium phenyl phosphate effect 60min with every 100mL serum.
The mensuration of alkaline phosphatase (AKP) adopts the disodium phenyl phosphate method., produce 1mg phenol person and be defined as an enzyme activity unit at 37 ℃ and substrate disodium phenyl phosphate effect 15min with every 100mL serum.
3, the mensuration of water quality
Ammoniacal nitrogen is received oxidizing process with hypobromous acid and is measured (GB12763.4-91);
Nitrite nitrogen is with diazonium-azo spectrphotometric method for measuring (GB12763.4-91)
4, the mensuration of vibrios sum
Detect the vibrios sum with the TCBS medium
Concrete steps are: with phosphate-buffered salt (PBS) institute's water sampling is carried out 10 dilutions, to 10
-4Get the dilution (10 of stoste and each concentration
-1, 10
-2, 10
-3, 10
-4) 0.5mL, to coat on the TCBS solid culture medium, the vibrios sum in the water body is calculated in colony counting.
The mensuration result of the growth indexes of sea cucumber, immunocompetence index and water quality situation sees table one～table four: from table, can find out; A forms that motility rate, rate of body weight gain, the long growth rate of body, acid phosphatase (ACP), alkaline phosphatase (AKP) best result do not reach 82.14%, 830.56%, 274.65%, 0.031U/mL, 0.034U/mL, and vibrios adds up to 6.78 * 10
2CFU/mL; Compare with industrial A group, C, D, the survival rate of E group, rate of body weight gain, the long growth rate of body, acid phosphatase (ACP), alkaline phosphatase (AKP) activity all have tangible rising, and the vibrios sum obviously reduces.Explanation is under the breeding way of routine, and not changing water can influence the survival rate of sea cucumber, rate of body weight gain, body long growth rate, acid phosphatase (ACP) and alkaline phosphatase (AKP) vigor.Under identical exchange water cycle, along with the concentration of Bdellovibrio telotroch raises, survival rate, rate of body weight gain, the long growth rate of body, acid phosphatase and alkaline phosphatase activity are in gradually rising, and the vibrios sum obviously reduces; The concentration that the Bdellovibrio telotroch is described is high more, and the immunity of sea cucumber is strong more, and survival rate is high more, and Expectancy is more little.B1-1～B8-1 ... Survival rate between D1-4～D8-4, rate of body weight gain, the long growth rate of body, acid phosphatase (ACP), alkaline phosphatase (AKP), vibrios sum be no significant difference almost, and explaining in water or in feed, adding concentration is 10～10
7Pfu/mL Bdellovibrio telotroch bacterium liquid; Water once to change (full pond) in 5-40 days; The long growth rate of survival rate of seedling, rate of body weight gain, body, acid phosphatase (ACP), alkaline phosphatase (AKP), the influence of vibrios sum to sea cucumber are little, all can reach and improve survival rate and immunity, promote the purpose of growth.
Through measuring, each item water-quality determination value in the A group pond all is higher than experiment group B, C, D group.Wherein, the testing result of A group ammoniacal nitrogen is 0.279mg/L, is higher than 0.124～0.211mg/L of B group, 0.129～0.234mg/L of C group, 0.096～0.199mg/L of D group; The testing result of the nitrite nitrogen of A group is 0.077mg/L, is higher than 0.025～0.057mg/L of B group, 0.030～0.061mg/L of C group, 0.018～0.035mg/L of D group.Can know that by above interpretation of result application concentration is 10-10
7Pfu/mL Bdellovibrio telotroch bacterium liquid can effectively improve the breeding water body environment, regulates the content of ammoniacal nitrogen, nitrite nitrogen in the water, thereby provides the water body environment that is beneficial to sea cucumber existence, and in water body and feed, adds best results simultaneously.
This shows no matter in water or in feed, adding concentration is 10～10
7The Bdellovibrio telotroch bacterium liquid of pfu/mL changed (full pond) water in 40 days and once can both reach the immunity and the survival rate that improve sea cucumber, promoted growth, and amount of vibrio in the controlled water, improved water quality.In water with in the feed, add simultaneously, effect is better.Promptly can descend water resource frugally, the protection environment can be saved cost again, increases economic efficiency, and promotes the fast development of aquaculture.
The mensuration of the growth indexes of table one sea cucumber, immune indexes and vibrios sum
The mensuration of the growth indexes of table two sea cucumber, immune indexes and vibrios sum
The mensuration of the growth indexes of table three sea cucumber, immune indexes and vibrios sum
The mensuration of the growth indexes of table four seas ginseng, immune indexes and vibrios sum
The foregoing description is a preferred implementation of the present invention; But embodiment of the present invention is not restricted to the described embodiments; Other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; All should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (8)
1. water-saving holothurian culture method is characterized in that comprising following operating procedure:
(1) gets free of contamination deep-sea, marine site water as breeding water; Breeding water injects and cultures the pond after sand filtration, deposition and sterilization; Culture the corrugated plating of placing in the pond through sterilization;
(2) in culturing the pond, add Bdellovibrio telotroch bacterium liquid, and strengthen aeration, stir, make the concentration of the Bdellovibrio telotroch of Chi Shuizhong reach 10～10
7Pfu/mL;
(3) in culturing the pond, throw in the sea cucumber seedling;
(4) culture during normal aeration, the control water temperature is at 15～30 ℃, the salinity of Chi Shui remains on 23 ‰～30 ‰, daily ration of feeding be the sea cucumber body weight mass percent 3～5%, bait is before input, using concentration is 10～10
7The Bdellovibrio telotroch bacterium liquid of pfu/mL soaks 15～30min; Whenever changed Chi Shui once, change at every turn and add Bdellovibrio telotroch bacterium liquid behind the water, make the Bdellovibrio telotroch concentration of Chi Shuizhong reach 10～10 at a distance from 5～40 days
7Pfu/mL;
The said input of step (3) sea cucumber seedling when 15～30 ℃ of water temperatures according to following standard: when sea cucumber seedling specification was 400/kg, throwing in density was 600/m
2When sea cucumber seedling specification was 200/kg, input density was 400 tails/m
2When sea cucumber seedling specification was 100/kg, input density was 300 tails/m
2
2. a kind of water-saving holothurian culture method according to claim 1 is characterized in that: the said breed of step (1) pond cleaning and sterilizing before injecting breeding water is clean; 3m in the said breed pond
2～4m
2Arrange 1 gas exhausting stone.
3. a kind of water-saving holothurian culture method according to claim 1 is characterized in that: the method for the said adding in step (2) and (4) Bdellovibrio telotroch bacterium liquid is: behind the Chi Shui dilution bacterium liquid of culturing in the pond, and the full pond of evenly splashing.
4. a kind of water-saving holothurian culture method according to claim 1 is characterized in that: the aeration rate of the said increasing aeration of step (2) is 90～100L/h/m
3The aeration rate of the said normal aeration of step (4) is 70～80L/h/m
3
5. a kind of water-saving holothurian culture method according to claim 1 is characterized in that: the said bait of step (4) is manufactured feed, and every day, bait throwing in was twice; Whenever adopted the soil pick-up at the bottom of siphonage is cultured the pond at a distance from 5～7 days.
6. a kind of water-saving holothurian culture method according to claim 1; It is characterized in that: step (2) and (4) said Bdellovibrio are Bdellovibrio (Bdellovibrio sp.) BDM01; Said Bdellovibrio is preserved in the Chinese typical culture collection center in the Wuhan University of Luojia Mountain, Wuhan, Hubei Province on April 28th, 2008, and deposit number is CCTCC NO:M208066.
7. a kind of water-saving holothurian culture method according to claim 1 is characterized in that: step (2) and (4) said Bdellovibrio telotroch bacterium liquid prepare according to following method: inoculation 0.5mL concentration is 10
7Cfu/mL Aeromonas hydrophila (Aeromonas hydrophila) is cultivated 20h for 26～30 ℃ in the nutrient broth liquid nutrient medium, culture fluid is behind the centrifugal 10～20min of 5000～8000rpm; After deposition suspends with 1mL physiological saline; Be added in the DNB liquid nutrient medium, insert Bdellovibrio BDM01 again, cultivate 36～48h in 28 ℃; Culture fluid is following behind the centrifugal 15～20min of 6000～8000rpm at 4 ℃; The supernatant that will contain the Bdellovibrio telotroch is behind the centrifugal 15～20min of 16000～18000rpm, and deposition use DNB liquid nutrient medium, water, physiological saline or 0.2mol/L pH value are 7.2～7.6 phosphate buffer suspension, and obtaining concentration is 10
9The Bdellovibrio telotroch bacterium liquid of cfu/mL.
8. a kind of water-saving holothurian culture method according to claim 7; It is characterized in that: said DNB liquid nutrient medium is that nutrient broth 0.8g, caseinic acid hydrolysate 0.5g and yeast extract 0.1g are dissolved in the 1000ml distilled water, regulates pH value to 7.2～7.6.
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