CN101933480A - Water-saving marine fish seedling culture method - Google Patents
Water-saving marine fish seedling culture method Download PDFInfo
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Abstract
The invention discloses a water-saving marine fish seedling culture method. The method comprises the following steps of: (1) selecting parent fishes; (2) intensively culturing the parent fishes; (3) taking eggs, fertilizing the parent fishes and breeding the fertilized eggs; (4) arranging a pond; (5), feeding; and (6) growing the seedlings. In the invention, a scheme which combines the prevention and the adjustment of Bdellovibrio nectophores and the environment regulation is adopted; Bdellovibrio nectophore liquid with a concentration of 10-107pfu/mL is added into the seedling growing pond, and meanwhile, the feed is socked for 15-30min in the Bdellovibrio nectophore liquid with a concentration of 10-107pfu/mL; thus, the average seedling survival percent is not less than 50%, the albino rate is reduced to 4.1%, and the aberration rate is reduced to 2.3%; in addition, the pathogenic or potential pathogenic bacteria in the water can be lysed to improve the water quality so that water can be changed after 40 days; therefore, the cost is saved, and the economic benefit is greatly improved. The water-saving method for growing seedlings of marine fishes is a new green and low-carbon pollution-free seedling growing method.
Description
Technical field
The invention belongs to seawater fish seedling growing process field, particularly a kind of raising turbot fry breeding rate and immunity promote that turbot is grown, improved water quality fast, saves water and energy, the non-harmful marine fish new method for raising seedling of green low-carbon.
Background technology
Turbot (Scophthalmus maximus) is a kind of famous and precious economic fish that originates in Europe, belongs to the meat-eating race left-eyed flounder of dwelling at the bottom of the seawater.Europe has reached industrialization breed level, its personality docility, and residual stinging adapts to low water temperature life seldom mutually, and growth is easily accepted manufactured feed rapidly, is easy to intensive culture.Artificial breeding success in 1999, at present domestic in Shandong, Hebei, Liaoning formed the breed climax along Bohai Sea Area, the artificial seed is in great demand, survival rate of seedling is generally on the low side at present, even at the commercial nursery of European countries, survival rate of seedling also has only 20～30%.Domestic manufacturer's survival rate of seedling is lower, is seriously restricting the development of turbot aquaculture.
Turbot is a little less than being highly brittle alevin stage, and there are serious quality problems in seed: albefaction rate, abnormal rate, abnormal lethality are high, and disease frequently takes place.Turbot is very sensitive to bacterium, and bacteriosis is the maximum disease of harm alevin stage, and the difficulty of turbot fry breeding mainly is that opening initial stage and prelarva " open fish glue ".Have greater loss during opening fish glue, lethality is more than 50%.
The bacteriosis infection velocity in seedling stage is fast, generally from finding that diseased individuals only takes 2～3 days to extensive morbidity.Sick seedling lethality height, if in time do not take suitable preventive measure, lethality can reach more than 90% sometimes.At present, mainly prevent and treat by antibiotics.Generally mainly use penicillin, tetracycline, chloramphenicol, wait prophylactic treatment and treatment with furazolidone, tetracycline, not only form pesticide resistance easily in the postlarva phase in alevin stage, and easily remaining in the fish body, the fry quality is descended.
" greenhouse+phreatic water is grown seedlings " is the most general pattern of present turbot fry breeding.This pattern of growing seedlings needs a large amount of water circulations, and the exhaustive exploitation of underground well water uses, and not only wastes the resource and the energy, causes the water resource exhaustion, also can have a strong impact on geologic aspects, for geological disaster hides some dangers for.A large amount of breeding wastewaters discharges, and also causes the severe contamination of ocean water body.
Bdellovibrio is the bacterium that a class has cracking ability.Causing a disease or potentially pathogenic organism on the one hand can the cracking water body, purifying water body, Bdellovibrio can be improved the aquaculture organism intestinal environment on the other hand, improves immunity, promotes seedling development by metamorphosis and growth, improves weed survival rate.
No matter existing studies show that as a kind of effective microorganism preparation, 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].
Up to the present, in the world also not to using the research report of Bdellovibrio telotroch in the turbot paedomorphosis process.
Summary of the invention
For overcoming above-mentioned the deficiencies in the prior art, the object of the present invention is to provide a kind of water-saving marine fish seedling-cultivating method; This seedling-cultivating method has reduced the turbot incidence of disease, abnormal rate and albefaction rate effectively, the survival rate height, and the seed quality is good, improves water quality, saves water and energy green low-carbon.
Purpose of the present invention realizes by following technical proposals: a kind of water-saving marine fish seedling-cultivating method comprises following operating procedure:
(1) selection of parent population:
From 2 ages of culturing select the adult turbot that the bodily form is complete, color and luster normal, healthy active, body surface is bright, fast growth and body weight at the parent of 1.7～2.2kg as parent population;
(2) parent population hot housing:
Parent population in 2: 1 ratio of male and female, is cultivated in the parent fish rearing pond; In cultivating the pond, add Bdellovibrio telotroch bacterium liquid, make the concentration of the Bdellovibrio telotroch of Chi Shuizhong reach 10～10
7Pfu/mL; Bait is 10～10 with concentration before drop into cultivating the pond
7The Bdellovibrio telotroch bacterium liquid of pfu/mL soaks 15～30min;
(3) adopt ovum, fertilization and hatching:
Turbot is gathered ovum and seminal fluid after hot housing reaches sexual maturity, carry out artificial insemination; The nursery pond of seawater fertilized egg putting into is equipped with; In nursery pond, add Bdellovibrio telotroch bacterium liquid, make the initial concentration of the Bdellovibrio telotroch of Chi Shuizhong reach 10～10
7Pfu/mL; Fertilized egg is through hatching newly hatched larvae in 3～5 days;
(4) cloth pond:
In training seedling pond, add Bdellovibrio telotroch bacterium liquid, make the Bdellovibrio telotroch concentration of Chi Shuizhong reach 10～10
7Pfu/mL; Newly hatched larvae is put into training seedling pond cultivate, the cloth pond density of newly hatched larvae is 2～30,000/m
3
(5) bait throwing in:
The feed of throwing something and feeding comprises wheel animalcule, halogen worm naupiar larva, halogen worm and pellet; Feed is 10～10 with concentration before dropping into training seedling pond
7The Bdellovibrio telotroch bacterium liquid of pfu/mL soaks 15～30min;
(6), make the concentration of the mixture of Bdellovibrio telotroch and leech plastid in the water of pond reach 10～10 every changing the mixture bacterium liquid that adds Bdellovibrio telotroch and leech plastid after Chi Shui (full pond) once, changes (full pond) water again in 5～40 days
7Pfu/mL; Fry enters forming stage through 80～90 days raising.
The described parent population of step (2) is preferably preferred healthy from the breeding process of import fry, the fry that grows fast is as parent population.
Water in the described cultivation of step (2) pond is through fine filtering, ultraviolet sterilization; Described bait is thrown something and fed 3 times every day; Water temperature is controlled at 19～20 ℃ in the described parent fish pond, and illumination is regulated and control at 1500～2500lux, and light application time is: preceding 2 9～11 hours every days of week, 19～20 hours every days after two weeks, through 2～3 months warm light regulation and control, parent population reached sexual maturity.
The described nursery pond of step (3) is for being the long 4m that is, wide is 3m, is the rectangle cement pit of 1.5m deeply, and 6 gas stones are set in the pond, little inflation.
The water temperature of cultivating in the seed process in the described training seedling of step (4) pond is 18～20 ℃, and salinity is 26～28 ‰, and pH is 8.0～8.2, dissolved oxygen 5.0～7.0mg/L; Constantly inflation during the seed rearing, aeration quantity was controlled at 20～40L/h/m in preceding 10 days
2, prelarva increases aeration quantity to 50～70L/h/m after opening fish glue gradually
2
The described bait throwing in of step (5) is the 9th～15 age in days wheel animalcule of throwing something and feeding, and the density of throwing something and feeding is 10/mL; The 10th～25 age in days halogen worm naupiar larva of throwing something and feeding, the density of throwing something and feeding is for by 0.2/mL; The 20th～35 age in days halogen worm of throwing something and feeding, the density of throwing something and feeding is for to 1/mL; The 35th age in days pellet of throwing something and feeding later on, the feeding volume of pellet is 1g/mL.
Step (2)～(5) described Bdellovibrio is Bdellovibrio (Bdellovibrio sp.) BDFM05, and described Bdellovibrio is preserved in Chinese typical culture collection center on August 7th, 2009, and deposit number is CCTCC NO:M 209172.
Described Bdellovibrio BDFM05 is carried out carrying out morphologic observation: BDFM05 after the negative staining under electron microscope be unicellular, oval, size is 1.43 * 0.53um, end is given birth to flagellum, and the flagellum length described Bdellovibrio BDFM05 of 2um is at least cultivated the transparent circular plaque that can form diameter 2～3mm in three days with the double-layer plate method in 28 ℃.
Described Bdellovibrio telotroch bacterium liquid prepares in accordance with the following methods: inoculation Aeromonas hydrophila (Aeromonas hydrophila, purchase in Guangdong Province DSMZ of Institute of Micro-biology, numbering GIM1.172) in the nutrient broth liquid nutrient medium, cultivate 20h for 26～30 ℃, culture fluid is behind the centrifugal 10～20min of 5000～8000rpm, precipitation is added in the DNB liquid nutrient medium, insert Bdellovibrio BDFM05, cultivate 36～48h for 28 ℃, culture fluid obtains precipitation and supernatant behind the centrifugal 15～20min of 6000～8000rpm; Again with supernatant behind the centrifugal 15～20min of 16000～18000rpm, precipitation is the suspension of 7.2～7.6 phosphate buffer with DNB liquid nutrient medium, water, physiological saline or 0.2mol/L pH value, obtains Bdellovibrio telotroch bacterium liquid.
Described 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.
The present invention has following advantage and effect with respect to prior art:
(1) the Bdellovibrio telotroch is meant the growth forms before Bdellovibrio is invaded host bacteria.It is very fast that the Bdellovibrio telotroch is invaded the process of host bacteria, can finish in general several seconds.For Bdellovibrio leech plastid,, kill pathogenic bacteria with the Bdellovibrio telotroch and have rapid-action, energetic advantage though exist the weakness that preparation is loaded down with trivial details, be difficult for preservation.
(2) add Bdellovibrio telotroch bacterium liquid in water and in the bait during parent fish rearing of the present invention, promoted nutrient absorption of parent population and growth fast, guarantee that gonad development is normal, cultivate good fertilized egg.
(3) the present invention adopts the scheme that the prevention of Bdellovibrio telotroch and adjusting, environment conditioning combine, and average survival rate of seedling reaches more than 48%, and albefaction rate reduces to 3%, abnormal rate reduces to 2%, the longest can changing in 40 days (full pond) water has once been saved seedling cost, has improved economic benefit.
(4) the present invention strengthens the regulation and control of environmental factors such as water quality, water temperature, illumination in larval stages, and the certain density Bdellovibrio telotroch bacterium liquid of input in water and in the feed, improved water quality, improved the immunity of survival rate of seedling and seedling, reduce albefaction rate and abnormal rate, prevented the generation of disease.
(5) Bdellovibrio telotroch of the present invention safety in the application of turbot is good: the method that the Bdellovibrio telotroch is eliminated pathogenic bacteria by cracking is a biological method, and is green, safe, pollution-free.Bdellovibrio can be infected, the characteristic of cracking host bacteria makes it to be suitable as the biological cleaning factor of restraining or removing pathogenic bacteria in organism and the environment thereof, and it is behind the intact host bacteria of cracking, can wither away automatically because of hungry, thereby overcome the side effect that the antibiotic abuse brings.Existingly studies show that Bdellovibrio, to human no pathogenicity to the fish free of toxic effects.
Coming breeding with the Bdellovibrio telotroch is a kind of promising new method.
Embodiment
Below in conjunction with embodiment the present invention is done further detailed description, but embodiments of the present invention are not limited thereto.
Embodiment 1: improve the new cultural method of turbot fry breeding survival rate, concrete steps are as follows:
1, parent population is selected:
Shandong Province plant, select from 2 turbot in age (Scophthalmus maximus) adult fishes of 60000 breed that the bodily form is complete, color and luster normal, healthy active, body surface is bright, fast growth, body weight 1000 of the parents of 1.5～2.0kg as parent population.Male and female are reasonably combined, to avoid close fertilization, prevent the germplasm degeneration.
2, parent population reinforced cultivating
Parent population carries out preferably according to 2: 1 ratios of male and female.Parent population is at 35m
2The cultivation pond in carry out hot housing, average 4/m
2After cultivating the water process secondary sand filtration earlier in the pond, use behind the violent aeration.Add Bdellovibrio telotroch bacterium liquid, make that Bdellovibrio telotroch concentration reaches 10,10 respectively in the water body
3, 10
5, 10
7Pfu/mL.Add necessary lecithin of parent population gonad development and unsaturated fatty acid in the used manufactured feed simultaneously.Before the bait throwing in, feeds utilized will be 10,10 with concentration
3, 10
5, 10
7The Bdellovibrio telotroch bacterium liquid of pfu/mL soaks 30min, and bait throwing in is is in batches thrown something and fed 3 every day.
Temperature and illumination control: water temperature is controlled at 19～20 ℃ in the parent fish pond, the illumination regulation and control are at 2000lux, light application time is: preceding 2 9～11 hours every days of week, 19～20 hours every days after two weeks, under suitable illumination regulation and control and fortification breeding condition, add the effect of Bdellovibrio telotroch, the parent population gonad development is accelerated, through 2 months warm light regulation and control, parent population can reach the ripe essence of producing and lay eggs.The Bdellovibrio telotroch has promoted nutrient absorption of parent population and growth fast, guarantees that gonad development is normal, cultivates good fertilized egg.
3, adopt ovum, fertilization, hatching
Turbot is cultivated under these conditions to sexual maturity, male and female is separated difference collecting semen and ovum, artificial insemination.After fertilization, the ovum that will rise and fall are separately hatched in (fertilized egg) 1000g of will the hatching immigration incubator box.Fertilized egg is hatched at 14 ℃, finishes hatching through 3 days time, obtains just to incubate young seedling.
4, the processing of water for larval nursing
In nursery pond, inject seawater through sand filtration, sedimentation, reach effective water level after, adds an amount of Bdellovibrio telotroch bacterium liquid, and the increasing aeration, mix, make that Bdellovibrio telotroch concentration reaches 10,10 respectively in the water body
3, 10
5, 10
7Pfu/mL.
5, Bu Chi
Newly hatched larvae is put into training seedling pond cultivate, the cloth pond density of newly hatched larvae is 20,000/m
3, the water temperature in the seed rearing process is 18～20 ℃, and salinity is 26～28 ‰, and pH is 8.0～8.2, dissolved oxygen 5.0～7.0mg/L, constantly inflation during the seed rearing, aeration quantity was controlled at 20-40L/h/m in preceding 10 days
2, prelarva increases aeration quantity to 50～70L/h/m after opening fish glue gradually
2
6, bait throwing in
The feed of throwing something and feeding comprises wheel animalcule, halogen worm naupiar larva, halogen worm and pellet.9～15 ages in days wheel animalcule of throwing something and feeding, the density of throwing something and feeding is 10/mL, 10～25 ages in days halogen worm naupiar larva of throwing something and feeding, the density of throwing something and feeding is 0.2/mL; 20～35 ages in days halogen worm of throwing something and feeding, the density of throwing something and feeding is 1/mL; The 35th age in days pellet of throwing something and feeding later on, the feeding volume of pellet is 1g/mL; Will throw diligent the throwing less, having enough gets final product.Before the bait throwing in, bait is 10,10 with concentration respectively all
3, 10
5, 10
7The Bdellovibrio telotroch bacterium liquid of pfu/mL soaks 30min.
7, management
Temperature, illumination between nursery stage are controlled, observe at any time, in time remove the sediment of bottom, pond and the foreign material of water surface, and the time-division seedling.Test is provided with the different Quan Chi pond water frequencies of changing: changed in 5 days water once, changed in 10 days water once, changed in 15 days water once, changed in 20 days water once, changed in 25 days water once, changed in 30 days water once, changed in 35 days water once, changed water once in 40 days, add Bdellovibrio telotroch bacterium liquid after changing (full pond) water at every turn, make the cell concentration in the water body reach 10～10
7Pfu/mL.Adding the bacterium method is: behind corresponding Chi Shui dilution bacterium liquid, and the full pond of evenly splashing, and strengthen aeration, thus bacterium liquid is mixed.80-90 days raising of fry process can enter forming stage.
8, test result analysis
Test is grouping in the following manner, measures survival rate, albefaction rate, abnormal rate and immunocompetence index and the water quality situation of each group when off-test respectively;
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 as follows:
A group: factory's flowing water pattern of growing seedlings, the normal feed of throwing something and feeding;
The B group: the Bdellovibrio telotroch bacterium liquid of splashing in water body makes its concentration reach 10,10 respectively
3, 10
5, 10
7Pfu/mL, the pellet of throwing something and feeding soaks without Bdellovibrio;
C group: the Bdellovibrio of not splashing in the water body, but the pellet of throwing something and feeding is for being 10,10 through concentration
3, 10
5, 10
7After the Bdellovibrio telotroch bacterium liquid of pfu/mL soaks;
The D group: the Bdellovibrio telotroch bacterium liquid of splashing in water body makes cell concentration reach 10,10 respectively
3, 10
5, 10
7Pfu/mL, throwing something and feeding through concentration simultaneously respectively is 10,10
3, 10
5, 10
7Pellet 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;
B1-2, 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;
The assay method of fish body immune indexes is as follows:
1, antalzyme activity measure serum lysozyme active by the turbidity colorimetric method for determining (Ellis A E.Lysozyme assays.[M] //Stolen J S, Fletcher T C, Anderson D P, etal.Techniques in Fish Immunology I.USA:SOS Publications, 1990:101-103.) under the experiment condition, it is 1 unit of enzyme activity (U) that the every min light absorption value of every mL serum reduces 0.001.
2, the mensuration of phenol oxidase (PO) vigor
Phenoloxidase activity is measured (Hern á ndez-L ó pez J such as adopting Hern á ndez-L ó pez, Gollas-Galvan T.Vargas-Albores F.Activation of the prophenoloxidase system ofthe brown Penaeus californiensis Holmes[J] .Comp Biochem Physiol, method 1996.113C:61-66.).Under experiment condition, the every min extinction of every mL serum increases by 0.001, is defined as 1 unit of enzyme activity (U).
3, superoxide dismutase (SOD) vigor
Superoxide dismutase (SOD) vigor advances by improved 1,2,3,-thrihydroxy-benzene autoxidation methods such as Deng Biyu.(Deng Biyu, Yuan Qinsheng, Li Wenjie. the 1,2,3,-thrihydroxy-benzene autoxidation of improvement is measured the method [J] of superoxide dismutase activity. biochemistry and biophysics progress, 1991,18 (2): 163～163)
The measurement result of survival rate, albefaction rate, abnormal rate and immunocompetence index sees Table one～table four: as can be seen from the table, A forms that motility rate, antalzyme activity, superoxide dismutase activity, phenol oxidase vigor best result do not reach 26.38%, 1.666U/mL, 100.06U/mL, 1.867U/mL, and albefaction rate and abnormal rate are minimum to be respectively 9.52% and 9.14%.Compare with industrial A group, survival rate, antalzyme activity, superoxide dismutase activity, the phenol oxidase vigor of B, C, D group significantly improve, and albefaction rate, abnormal rate obviously reduce.Explanation is under the seedling raising manners of routine, and not changing water can influence the survival rate of turbot, antalzyme activity, superoxide dismutase activity, phenol oxidase vigor.Under identical exchange water cycle, along with Bdellovibrio telotroch concentration raises, survival rate, antalzyme activity, superoxide dismutase activity, phenol oxidase vigor are all raising gradually, and albefaction rate, abnormal rate obviously reduce; The concentration that the Bdellovibrio telotroch is described is high more, and the immunity of turbot is strong more, and survival rate is high more, and Expectancy is more little.B 1-1～B8-1 ... survival rate between D1-4～D8-4, antalzyme activity, superoxide dismutase activity, phenol oxidase vigor, albefaction rate, abnormal rate no significant difference, illustrating in water or add concentration in feed is 10～10
7Pfu/mL Bdellovibrio telotroch bacterium liquid changed (full pond) water in 5～40 days once, and is not obvious to survival rate of seedling, antalzyme activity, superoxide dismutase activity, phenol oxidase vigor, albefaction rate, abnormal rate influence, all can improve survival rate and immunity.
Result of the test shows that the seedling raising manners of B, C, D group all can improve the immunity of survival rate of seedling and seedling, promote to ingest and growth fast, and D group best results.That is: no matter adding concentration in water or in feed is 10～10
7Pfu/mL Bdellovibrio telotroch bacterium liquid reaches 40 days and does not change water, all can obviously improve the immune indexes such as survival rate of seedling, antalzyme activity, superoxide dismutase activity, phenol oxidase vigor of turbot, reduces albefaction rate and abnormal rate; Add simultaneously in water and in the feed, effect is better.Change (full pond) water in 5～40 days and once antalzyme activity, superoxide dismutase activity, phenol oxidase vigor, albefaction rate, abnormal rate are not all had obvious influence.
The total number of bacteria of A group and vibrios sum are respectively up to 1.63 * 10
3Cfu/mL and 1.13 * 10
2Cfu/mL.The total number of bacteria of B, C, D group is respectively 1.58 * 10
2Cfu/mL, 1.64 * 10
2Cfu/mL, 1.11 * 10
2Cfu/mL; Vibrios sum 6.5 * 10cfu/mL, 7.3 * 10cfu/mL, 4.2 * 10cfu/mL with A group ratio, have reduced by 1 order of magnitude.The PH of A, B, C, D group between 7.79～8.10, changes little.
This shows no matter adding concentration in water or in feed is 10～10
7The Bdellovibrio telotroch bacterium liquid of pfu/mL all can improve the immunity and the survival rate of seedling of seedling, and can control bacterial number in the water, improves water quality.Add simultaneously in water and in feed, 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.
Table one survival rate, albefaction rate, abnormal rate and immunocompetence index
Table two survival rate, albefaction rate, abnormal rate and immunocompetence index
Table three survival rate, albefaction rate, abnormal rate and immunocompetence index
Table four survival rate, albefaction rate, abnormal rate and immunocompetence index
The foregoing description is a preferred implementation of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spiritual essence 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 marine fish seedling-cultivating method is characterized in that comprising following operating procedure:
(1) selection of parent population:
Selective body focuses on the parent of 1.5～2.0kg as parent population from the 2 age adult turbots of culturing;
(2) parent population hot housing:
Parent population in 2: 1 ratio of male and female, is cultivated in the parent fish rearing pond; In cultivating the pond, add Bdellovibrio telotroch bacterium liquid, make the concentration of the Bdellovibrio telotroch of Chi Shuizhong reach 10～10
7Pfu/mL; Bait is 10～10 with concentration before drop into cultivating the pond
7The Bdellovibrio telotroch bacterium liquid of pfu/mL soaks 15～30min;
(3) adopt ovum, fertilization and hatching:
Turbot is gathered ovum and seminal fluid after hot housing reaches sexual maturity, carry out artificial insemination; The nursery pond of seawater fertilized egg putting into is equipped with; In nursery pond, add Bdellovibrio telotroch bacterium liquid, make the concentration of the Bdellovibrio telotroch of Chi Shuizhong reach 10～10
7Pfu/mL; Fertilized egg is through hatching newly hatched larvae in 3～5 days;
(4) cloth pond:
In training seedling pond, add Bdellovibrio telotroch bacterium liquid, make the concentration of the Bdellovibrio telotroch of Chi Shuizhong reach 10～10
7Pfu/mL; Newly hatched larvae is put into training seedling pond cultivate, the cloth pond density of newly hatched larvae is 2～30,000/m
3
(5) bait throwing in:
The feed of throwing something and feeding comprises wheel animalcule, halogen worm naupiar larva, halogen worm and pellet; Feed is 10～10 with concentration before dropping into training seedling pond
7The Bdellovibrio telotroch bacterium liquid of pfu/mL soaks 15～30min;
(6) changed Chi Shui once every 5～40 days, add Bdellovibrio telotroch bacterium liquid again after changing water, make the concentration of Bdellovibrio telotroch in the water of pond reach 10～10
7Pfu/mL; Fry enters forming stage through 80～90 days raising.
2. a kind of water-saving marine fish seedling-cultivating method according to claim 1 is characterized in that: the water in the described cultivation of step (2) pond is through fine filtering; Described bait is thrown something and fed 3 times every day; Water temperature is controlled at 19～20 ℃ in the described parent fish pond, and illumination is regulated and control at 1500～2500lux, and light application time is: preceding 2 9～11 hours every days of week, 19～20 hours every days after two weeks, through 2～3 months warm light regulation and control, parent population reached sexual maturity.
3. a kind of water-saving marine fish seedling-cultivating method according to claim 1 is characterized in that: the described nursery pond of step (3) is the long 4m that is, wide is 3m, is the rectangle cement pit of 1.5m deeply, and 6 gas stones are set in the pond, little inflation.
4. a kind of water-saving marine fish seedling-cultivating method according to claim 1 is characterized in that: the water temperature of cultivating in the seed process in the described training seedling of step (4) pond is 18～20 ℃, and salinity is 26～28 ‰, and pH is 8.0～8.2, dissolved oxygen 5.0～7.0mg/L; Constantly inflation during the seed rearing, aeration quantity was controlled at 20～40L/h/m in preceding 10 days
2, prelarva increases aeration quantity to 50～70L/h/m after opening fish glue gradually
2
5. a kind of water-saving marine fish seedling-cultivating method according to claim 1 is characterized in that: the described bait throwing in of step (5) is the 9th～15 age in days wheel animalcule of throwing something and feeding, and the density of throwing something and feeding is 10/mL; The 10th～25 age in days halogen worm naupiar larva of throwing something and feeding, the density of throwing something and feeding is for by 0.2/mL; The 20th～35 age in days halogen worm of throwing something and feeding, the density of throwing something and feeding is for to 1/mL; The 35th age in days pellet of throwing something and feeding later on, the feeding volume of pellet is 1g/mL.
6. a kind of water-saving marine fish seedling-cultivating method according to claim 1, it is characterized in that: step (2)～(5) described Bdellovibrio is Bdellovibrio (Bdellovibrio sp.) BDFM05, described Bdellovibrio is preserved in Chinese typical culture collection center on August 7th, 2009, and deposit number is CCTCC NO:M 209172.
7. a kind of water-saving marine fish seedling-cultivating method according to claim 1, it is characterized in that: described Bdellovibrio telotroch bacterium liquid prepares in accordance with the following methods: inoculation Aeromonas hydrophila (Aeromonashydrophila) is in the nutrient broth liquid nutrient medium, cultivate 20h for 26～30 ℃, culture fluid is behind the centrifugal 10～20min of 5000～8000rpm, precipitation is added in the DNB liquid nutrient medium, insert Bdellovibrio BDFM05, cultivate 36～48h for 28 ℃, culture fluid obtains precipitation and supernatant behind the centrifugal 15～20min of 6000～8000rpm; Again with supernatant behind the centrifugal 15～20min of 16000～18000rpm, precipitation is the suspension of 7.2～7.6 phosphate buffer with DNB liquid nutrient medium, water, physiological saline or 0.2mol/L pH value, obtains Bdellovibrio telotroch bacterium liquid.
8. a kind of water-saving marine fish seedling-cultivating method according to claim 7, it is characterized in that: described 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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| CN1596628A (en) * | 2004-08-16 | 2005-03-23 | 海阳市黄海水产有限公司 | Fry culturing method for controlling albefaction rate of scophthalmus maximus |
| CN101275119A (en) * | 2008-01-24 | 2008-10-01 | 华南理工大学 | Application of bdellovibrio to eliminating pathogen in freshwater product and culture water thereof |
| CN101638629A (en) * | 2009-08-28 | 2010-02-03 | 华南理工大学 | Bdellovibrio bacteriovorus for preventing and curing rice bacterial diseases and application thereof |
| CN101695286A (en) * | 2009-10-13 | 2010-04-21 | 中国水产科学研究院黄海水产研究所 | Juvenile turbot culturing and feeding method |
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| CN1596628A (en) * | 2004-08-16 | 2005-03-23 | 海阳市黄海水产有限公司 | Fry culturing method for controlling albefaction rate of scophthalmus maximus |
| CN101275119A (en) * | 2008-01-24 | 2008-10-01 | 华南理工大学 | Application of bdellovibrio to eliminating pathogen in freshwater product and culture water thereof |
| CN101638629A (en) * | 2009-08-28 | 2010-02-03 | 华南理工大学 | Bdellovibrio bacteriovorus for preventing and curing rice bacterial diseases and application thereof |
| CN101695286A (en) * | 2009-10-13 | 2010-04-21 | 中国水产科学研究院黄海水产研究所 | Juvenile turbot culturing and feeding method |
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| 《水产科技情报》 20071231 刘朝阳等 噬菌蛭弧菌及其在大菱鲆养殖中的应用探讨 第271页左栏第1段至第273页左栏第1段 1-8 , 第06期 2 * |
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