CN102017909B - Energy-saving method for culturing scophthalmus maximus - Google Patents
Energy-saving method for culturing scophthalmus maximus Download PDFInfo
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Abstract
The invention discloses an energy-saving method for culturing scophthalmus maximus. The method comprises the following steps of: selecting a culturing water source; treating water in a pool; treating a feed; controlling a water changing period; and controlling a culturing environment by a bio-ecology method according to the bio-ecology principle of a culturing water body, the requirement of marine organisms on growth environments, mutual relation between organisms and a nuisanceless culturing standard. Key technology is that: bdellovibrio bacteriovorus plasmid bacterial liquid is added into awater body; the feed is soaked in the bdellovibrio bacteriovorus plasmid bacterial liquid; and the water in the pool can be changed once every 40 days in the culturing period. The method is a biological culturing method, which is environmentally-friendly, safe and pollutant-free, has remarkable effect and extremely good development prospect, can improve water quality, promotes ingestion of the scophthalmus maximus, enhances immunity, promotes growth and increases survival rate. The method plays an important role in enhancing economic benefit and eating safety and guaranteeing human health.
Description
Technical field
The invention belongs to the marine fish culture technical field, particularly a kind of raising turbot immunity promotes that turbot is grown, improved water quality fast, saves water and energy, uses manpower and material resources sparingly, the turbot cultural method of the novel non-pollution of green low-carbon.
Background technology
With the turbot (Scophthamus maximus) of " how precious fish " name as a kind of cold water mariculture kind of high-quality, adapt to low water temperature life, fast growth, personality docility, strong stress resistance, be fit to high-density breeding with it, have that colloid is abundant concurrently, delicious meat, mouthfeel uniqueness, can improve people's good characteristic such as immunity and be subjected to the welcome in numerous raisers and market deeply.Enter at present the restaurant, hotel etc. of big and medium-sized cities as famous and precious marine product.
Turbot is introduced China and is had nearly ten years, wherein carry out large-scale industrial aquaculture 5～6 years were also arranged, facts have proved that this kind is very suitable for the coastal development of special aquaculture industry of northern China, is the coastal breed breeding few in number in the current north.Yet along with the prolongation of culturing time and the continuous expansion of the scale of breed, the frequency that various diseases take place and scale are also constantly enlarging, and have increased raiser's production cost greatly, have reduced income, become the bottleneck of restriction aquaculture sustainable development.
" green house+phreatic water is cultured " is the most general production model of present aquaculture.This aquaculture model needs a large amount of water circulations, and the exhaustive exploitation of underground well water uses, and not only wastes 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.
Pathogenic infection is to cause the dead and poky main cause of turbot, and bacterium, fungi, virus, mycoplasma etc. are arranged.Wherein main pathogenic or potentially pathogenic organism is bacterium, and Vibrio anguillarum, vibrio parahaemolytious, vibrio alginolyticus, husky vibrios, Vibrio harveyi, Vibrio vulnificus etc. are multiple, and lethality can reach more than 50%.The infection of germ, the serious development that is restricting China's mariculture industry.Traditional antibiotic and the use of chemicals have absorption soon, eutherapeutic advantage, but can produce excitant to turbot, can cause new inflammation.Long-term use antibiotic can make and cause a disease or the increase of potentially pathogenic organism antibody-resistant bacterium simultaneously, and some common epidemic diseases are difficult to thorough radical cure, repeatedly outbreak.Have, antibiotic residue can influence the quality of fish again, harm people's healthy, harm ecotope, reduces its commercial value, also also causes simultaneously raiser's economic loss.Have drug residue free, cause a disease or potentially pathogenic organism can not develop immunity to drugs, economic benefit is high advantage with herbal control bacterium epidemic disease, but have take effect slow, the shortcoming that production efficiency is lower.
Therefore, promote that turbot is grown fast, saves water and energy, used manpower and material resources sparingly, the novel non-pollution breeding way of green low-carbon becomes the great demand of country in recent years, cause the extensive concern of aquatic products circle.At present, improve the immunity of turbot with Bdellovibrio leech plastid, promote the turbot breeding way report as yet at home and abroad of growth fast.
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].
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 energy-saving turbot cultural method; This method realizes by add Bdellovibrio leech plastid in feed and water body, specifically comprises the breed choosing source, the processing of Chi Shui, the processing of feed, the control of exchange water cycle, its feature is learned principle according to breeding ecological, halobiontic growing environment requirement, correlation between the biology and nuisanceless breed standard are manually created breeding environment, reduce input cost, improve water quality, improve turbot immunity, promote turbot to grow fast, increase economic efficiency.
Purpose of the present invention realizes by following technical proposals: a kind of energy-saving turbot cultural 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; Cover gobo above culturing the pond, the control intensity of illumination is 150～200lux;
(2) in culturing the pond, add Bdellovibrio leech plastid bacterium liquid, and strengthen aeration, stir, make the initial concentration of the Bdellovibrio leech plastid of Chi Shuizhong reach 10～10
7Pfu/mL;
(3) throw in fry to culturing in the pond;
(4) culture during normal aeration, the control water temperature is at 15～30 ℃, daily ration of feeding is the mass percent 3～5% of fish body weight, bait is 10～10 with concentration before input
7Pfu/mL Bdellovibrio leech plastid bacterium liquid soaks 15～30min; Changed Chi Shui (full pond) every 5～40 days once, add Bdellovibrio leech plastid bacterium liquid after changing water at every turn, make the Bdellovibrio leech plastid concentration of Chi Shuizhong reach 10～10
7Pfu/mL.
Described Bdellovibrio leech plastid is that Bdellovibrio is present in the state in the host bacterium;
Be matrix or flat at the bottom of the pond in the described breed of step (1) pond, breed pond cleaning and sterilizing before injecting breeding water is clean; The shading rate of described gobo is 95%.
The method of the described adding in step (2) and (4) Bdellovibrio leech plastid 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 described increasing aeration of step (2) is 90～100L/h/m
3The aeration rate of the described normal aeration of step (4) is 70～80L/h/m
3
The described input of step (3) fry when 15～30 ℃ of water temperatures according to following standard: when fry weight was 25～50g, input density was 300～400 tails/m
2When fry weight was 70～100g, input density was 200～300 tails/m
2When fry weight was 100～200g, input density was 100～150 tails/m
2Temperature then reduces cultivation density when raising.
The described bait of step (4) is granular bait for fish; Described bait in the morning, noon and evening each throw in once, each bait throwing in 5～6 hours at interval, the bait throwing in mode is for perfusing full pond.
Step (2) and (4) described Bdellovibrio are Bdellovibrio (Bdellovibrio sp.) BDFM05, described Bdellovibrio is preserved in Chinese typical culture collection center in the Wuhan University of Luojia Mountain, Wuhan, Hubei Province on August 7th, 2009, and deposit number is CCTCC NO:M 209172.
Described Bdellovibrio BDFM05 carries out carrying out morphologic observation: BDFM05 after the negative staining under electron microscope and is unicellular, ellipse, and size is 1.43 * 0.53um, and end is given birth to flagellum, and flagellum length is 2um at least; Described Bdellovibrio BDFM05 cultivates the transparent circular plaque that can form diameter 2～3mm in three days with the double-layer plate method in 28 ℃;
Described Bdellovibrio BDFM05 is obtained through ultraviolet mutagenesis by wild type Bdellovibrio BDS02;
Described ultraviolet mutagenesis is according to application number " 2008100220488.3 ", and the method that name is called in the national inventing patent application of " application of uv induction sudden change in strengthening the Bdellovibrio cracking performance " is carried out;
Described leech plastid is preferably used DNB liquid nutrient medium (nutrient broth 0.8g, caseinic acid hydrolysate 0.5g, yeast extract 0.1g is dissolved in the 1000mL distilled water, and the pH value is 7.2～7.6), water, physiological saline or 0.2mol/L pH value be that 7.2～7.6 phosphate buffer suspends;
Described Bdellovibrio leech plastid 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 to DNB liquid nutrient medium (nutrient broth 0.8g, caseinic acid hydrolysate 0.5g, yeast extract 0.1g, be dissolved in the 1000mL distilled water, the pH value is 7.2～7.6) in, insert Bdellovibrio BDFM05, cultivate 36～48h for 28 ℃, culture fluid is behind the centrifugal 15～20min of 6000～8000rpm, removal contains the supernatant of Bdellovibrio telotroch, precipitation DNB liquid nutrient medium, water, physiological saline or 0.2mol/L pH value are 7.2～7.6 phosphate buffer suspension, obtain Bdellovibrio leech plastid 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.
Advantage of the present invention and effect: developing nuisanceless breed, improve the immunity of turbot, promote the quick growth of turbot, reduce production costs, increase economic efficiency, is the road that must have of China's mariculture industry development.
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, produce a hole at the host cell wall, invade host's periplasmic space when Bdellovibrio after hole seal again.Telotroch loses flagellum and host cell is called as " leech plastid " together.
Compare with telotroch, the leech plastid not only has preparation, preservation is more convenient, and the environment tolerance is strong, though omit characteristics more of a specified duration slowly action time, also has potential inactivated vaccine function simultaneously.Behind Bdellovibrio telotroch invasion host cell, the respiration of host cell (respiration) is stopped very soon.Bdellovibrio also enters leech plastid state, begins to secrete plurality of enzymes, and the big molecule of clearing up host cell is small-molecule substance, and then is integrated into the nutriment of self, makes self growth, and is elongated.Finally, the leech plastid of strip is divided into the sections of a lot of weak points, grows flagellum, becomes telotroch again.Afterwards, the essential enzyme of they secretions, broken wall and going out becomes the telotroch that breaks away from host bacteria, has flagellum, free swimming again.Between this leech plastid puberty, Bdellovibrio does not change the surface texture of host cell, makes former host still have immunogenicity, but the stimulating organism body produces immune response.This potential function makes uses Bdellovibrio leech plastid in the breed of turbot, can effectively prevent the generation of bacteriosis, improves and cultures survival rate etc.
Bdellovibrio can be improved the aquaculture organism intestinal environment, improves immunity, promotes growth.Existingly studies show that Bdellovibrio to the fish free of toxic effects, to human no pathogenicity.
These advantages make the leech plastid be suitable as equally to restrain or remove potential causing a disease or the biological cleaning factor of potentially pathogenic organism in organism and the environment thereof.Aquatic products is caused a disease the leech plastid or potentially pathogenic organism, particularly vibrio marinopraesens have strong cracking strength, and water quality is also had certain catharsis, soaks feed with the leech plastid, and the certain density leech plastid bacterium liquid of splashing in the pool water within, the longlyest reaches 40 days and do not change water.Can promote the growth of turbot, improve immunity, can purify water again, reduce quantity of exchanged water, thereby overcome side effect that the antibiotic abuse brings and to breeding wastewater handle expensive, for the mankind provide safety, green, non-harmful aquatic products, improve raiser's income.
Embodiment
Do further detailed description below in conjunction with the present invention of embodiment, but embodiments of the present invention are not limited thereto.
Concrete enforcement of the present invention place is Shandong Province plant, and the enforcement time is 8 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, enter 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 turbot is built in and is cultured in the booth, and each sump area is 40m
2, 1.2 meters of the depth of waters are matrix at the bottom of the pond, be conducive to operations such as draining soil pick-up, but also can in flat pond.It is 95% gobo that pond top covers obscurity, avoids illumination strong excessively.Intensity of illumination is generally controlled at 150～200lux.
Select physical health, individual turbot of uniform size (Scophthalmus maximus) juvenile fish, average weight is 28 ± 2.5g, and average body length is 11 ± 0.55cm, adopts the randomized block experiment design, according to 300 tails/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 as follows:
A group: normal flowing water, the pellet of throwing something and feeding (breeding way of plant produced);
B group: the Bdellovibrio leech plastid bacterium liquid of splashing in water makes that the Bdellovibrio bacterial concentration reaches 10,10 respectively in the water body
3, 10
5, 10
7Pfu/mL throws in pellet;
The C group: the concentration of throwing something and feeding is 10,10
3, 10
5, 10
7Pellet after the Bdellovibrio leech plastid bacterium liquid of pfu/mL soaks;
D group: in water, add Bdellovibrio leech plastid bacterium liquid, cell concentration is reached 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
7Pellet after the Bdellovibrio leech plastid 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;
Particles used People's Republic of China's agricultural industry criteria " pollution-free food fishing manufactured feed safety limit " regulation (NY5072-2002) that meets.Duration of test, the mass fraction of daily ration of feeding and fish body weight is 3-5%, throwing something and feeding every day, three times (17:00), the process of at every turn throwing something and feeding continues 30min for 7:00,12:00.Growth indexes, immune indexes and the water quality situation of fish measured in off-test.
(1), the assay method of fish bulk-growth index is as follows:
Randomly draw 40 tail fishes for every group and detect growing state, length of fish body is measured and adopted precision is the slide measure tolerance of 1mm, and body is long holds the distance of vertebrae between bottom for the kiss of fish.Body weight adopts electronic balance weighing.The test growth indexes is calculated as follows:
Survival rate (%)=N
t/ N
o* 100%;
Rate of body weight gain (%)=(W
t-W
o)/W
o* 100%;
The long growth rate (%) of body=(L
t-L
o) L/
o* 100%;
Coefficient of condition=W
o/ L
iOr W
t/ L
e
N in the formula
oBe the total mantissa of on-test macrura reevesii body; N
tBe the total mantissa of off-test macrura reevesii body; W
oBe on-test macrura reevesii body weight; W
tBe off-test macrura reevesii body weight; L
oFish length during for on-test; L
tFish length during for off-test;
The every growth indexes of turbot that test records sees Table one～table four:
As can be seen from the table, A composition motility rate, rate of body weight gain, the long growth rate of body, coefficient of condition are respectively 80.28%, 1779%, 54.76%, 20.23, compare with the A group, B, C, the survival rate of D group, rate of body weight gain, the long growth rate of body, coefficient of condition all are significantly improved.Explanation is under the breeding way of routine, and not changing water can influence the survival rate of turbot, rate of body weight gain, the long growth rate of body, coefficient of condition.Under identical exchange water cycle, along with the rising of Bdellovibrio concentration, survival rate, rate of body weight gain, the long growth rate of body, coefficient of condition are all raising gradually, and are all organizing apparently higher than A.B1-1～B8-1 ... survival rate between D1-4～D8-4, rate of body weight gain, the long growth rate of body, coefficient of condition no significant difference.Concentration is added in explanation in water or in feed be 10～10
7Pfu/mL Bdellovibrio leech plastid bacterium liquid does not change water between 5-40 days, to survival rate, rate of body weight gain, the long growth rate of body, coefficient of condition influence not quite, can both reach the purpose that promotion is ingested and grown.
Therefore, no matter still adding concentration in the feed in water is 10～10
7Pfu/mL Bdellovibrio leech plastid bacterium liquid reaches and do not change water in 40 days and all can promote ingesting of turbot and growth fast, but both adds (D group), effect optimum simultaneously.
The detection of table one turbot growth indexes
The detection of table two turbot growth indexes
The detection of table three turbot growth indexes
The detection of table four turbot growth indexes
(2), the assay method of fish body immune indexes is as follows:
1, infection experiment
Infection experiment is carried out in the pond that fills 1 ton of sand filtration seawater (salinity 29～30 ‰) of sterilization in advance.From experiment A group, B1-1, B1-2 ... D8-3, D8-4 in totally 97 groups each group get 100 of turbot, body weight 500 ± 15g/ bar at random.Be 10 with aseptic 1mL syringe through turbot lumbar injection 0.5mL concentration
8Cfu/mL Aeromonas hydrophila (Aeromonas hydrophila purchases in Guangdong Province DSMZ of Institute of Micro-biology, numbering GIM1.172) infects turbot.The test water temperature remains on 18～20 ℃, and flowing water is cultured, and keeps 8 water exchange capacities every day, and inflation is cultivated.Behind the artificial infection, the body surface of record turbot changes, moves, ingests and death condition.Observed 10 days continuously, finally be calculated to be motility rate, the employing relative survival rate (Relative Percent Survival, RPS) calculating formula:
RPS (%)=(1-immune group lethality/control group lethality) * 100%
2, 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.
3, 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).
4, 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 immune indexes sees Table five～table eight:
As can be seen from the table, A group antalzyme activity, superoxide dismutase activity, phenol oxidase vigor best result Wei 18.56U/mL, 210.28U/mL, 6.08U/mL.Compare with the A group, antalzyme activity, superoxide dismutase activity, the phenol oxidase vigor of B, C, D group all are significantly improved.Explanation is under the breeding way of routine, and not changing water can influence the antalzyme activity of turbot, superoxide dismutase activity, phenol oxidase vigor.Under identical exchange water cycle, along with the concentration rising of Bdellovibrio leech plastid, antalzyme activity, superoxide dismutase activity, phenol oxidase vigor are all raising gradually.The concentration that Bdellovibrio leech plastid is described is more high, and the immunity of turbot is more strong.B1-1～B8-1 ... antalzyme activity between D1-4～D8-4, superoxide dismutase activity, phenol oxidase vigor no significant difference, illustrating in water or add concentration in feed is 10～10
7Pfu/mL Bdellovibrio leech plastid bacterium liquid does not change water between 5-40 days, little to antalzyme activity, superoxide dismutase activity, phenol oxidase effect of vigor, all can reach the purpose that improves immunity.
The result of infection experiment shows, relative survival rate (the Relative Percent Survival of A group, RPS) be 0, compare with the A group, the relative survival rate of B, C, D group all is significantly improved, as can be seen from Table 9, under identical exchange water cycle, rising along with Bdellovibrio leech plastid bacterial concentration, relative survival rate raises gradually, and all apparently higher than the A group of plant produced, the method that adopts novel interpolation Bdellovibrio leech plastid is described, can obviously improve the relative survival rate of turbot, namely improve the immune resistance of turbot; From different exchange water cycles, the turbot relative survival rate of respectively organizing under the same concentrations does not change significantly.Illustrate under the novel cultural method that do not change water in 5-40 days, Bdellovibrio leech plastid bacterium liquid can both strengthen the immunocompetence of turbot.With regard to B, C, three groups of different disposal modes of D, all can improve the relative survival rate of turbot, and in water body and feed, add best results simultaneously.
Therefore, no matter still add certain density Bdellovibrio leech plastid bacterium liquid in the feed in water, reach 40 days and do not change water, all can improve the immune resistance of turbot, add simultaneously in water with in the feed, effect is best.Thereby show that further Bdellovibrio leech plastid has potential vaccine effect, can stimulate the turbot immune response, resist the infringement of Aeromonas, help to improve the survival rate of turbot.
Result of the test shows that the breeding way of B, C, D group can reach the immunity that improves fish, the purpose that promotion is ingested and grown fast, and D group best results.That is: no matter adding concentration in water or in feed is 10～10
7Pfu/mL Bdellovibrio leech plastid bacterium liquid reaches 40 days and does not change water, all can significantly improve the immune indexes such as antalzyme activity, superoxide dismutase activity, phenol oxidase vigor of turbot, adds simultaneously in water and in the feed, and effect is better.
The total number of bacteria of A group and vibrios sum are respectively up to 2.95 * 10
3Cfu/mL and 5.42 * 10
2Cfu/mL.The total number of bacteria of B, C, D group is respectively 4.81 * 10
2Cfu/mL, 5.87 * 10
2Cfu/mL, 4.84 * 10
2Cfu/mL; Vibrios sum 8.9 * 10cfu/mL, 9.7 * 10cfu/mL, 4.3 * 10cfu/mL have reduced by 1 order of magnitude than the A group.The PH of A, B, C, D group is between 7.78～8.12.
This shows no matter adding concentration in water or in feed is 10～10
7The Bdellovibrio leech plastid bacterium liquid of pfu/mL all can reach the immunity that improves fish, the purpose that promotion is ingested and grown fast, and potential the causing a disease or potentially pathogenic organism in the splitting water, reaches the effect of improving water quality.Add simultaneously in water and in feed, effect is better.Can either save groundwater resources, the protection environment can be saved cost again, increases economic efficiency, and promotes the fast development of aquaculture.
The measurement result of table five immune indexes
The measurement result of table six immune indexes
The measurement result of table seven immune indexes
The measurement result of table eight immune indexes
Table nine is attacked malicious infection experiment relative survival rate
D group in above-described embodiment is 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 (2)
1. energy-saving turbot cultural method is characterized in that comprising 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 and post precipitation; Cover gobo above culturing the pond, the control intensity of illumination is 150～200lux;
(2) in culturing the pond, add Bdellovibrio leech plastid bacterium liquid, and strengthen aeration, stir, make the initial concentration of the Bdellovibrio leech plastid of Chi Shuizhong reach 10～10
7Pfu/mL;
(3) throw in fry to culturing in the pond;
(4) culture during normal aeration, the control water temperature is at 15～30 ℃, daily ration of feeding is the mass percent 3～5% of fish body weight, bait is 10～10 with concentration before input
7Pfu/mL Bdellovibrio leech plastid bacterium liquid soaks 15～30min; Changed Chi Shui once every 5～40 days, change at every turn and add Bdellovibrio leech plastid bacterium liquid behind the water, make the Bdellovibrio leech plastid concentration of Chi Shuizhong reach 10～10
7Pfu/mL;
The described input of step (3) fry when 15～30 ℃ of water temperatures according to following standard: when fry weight was 25～50g, input density was 300～400 tails/m
2When fry weight was 70～100g, input density was 200～300 tails/m
2When fry weight was 100～200g, input density was 100～150 tails/m
2
Described Bdellovibrio leech plastid bacterium liquid prepares in accordance with the following methods: the inoculation Aeromonas hydrophila 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 is behind the centrifugal 15～20min of 6000～8000rpm, removal contains the supernatant of Bdellovibrio telotroch, precipitation DNB liquid nutrient medium, water, physiological saline or 0.2mol/L and pH value are 7.2～7.6 phosphate buffer suspension, obtain Bdellovibrio leech plastid 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;
Described Bdellovibrio BDFM05 is preserved in Chinese typical culture collection center on August 7th, 2009, and deposit number is CCTCC NO:M 209172.
2. a kind of energy-saving turbot cultural method according to claim 1 is characterized in that: be matrix or flat at the bottom of the pond in the described breed of step (1) pond, it is clean to culture pond cleaning and sterilizing before injecting breeding water; The shading rate of described gobo is 95%.
3, a kind of energy-saving turbot cultural method according to claim 1 is characterized in that: the method for the described adding in step (2) and (4) Bdellovibrio leech plastid 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 energy-saving turbot cultural method according to claim 1, it is characterized in that: the aeration rate of the described increasing aeration of step (2) is 90～100L/h/m
3The aeration rate of the described normal aeration of step (4) is 70～80L/h/m
3
5, a kind of energy-saving turbot cultural method according to claim 1, it is characterized in that: the described bait of step (4) is granular bait for fish; Described bait in the morning, noon and evening each throw in once, each bait throwing in is spaced apart 5～6 hours, the bait throwing in mode is for perfusing full pond.
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| 刘朝阳 等.噬菌蛭弧菌及其在大菱鲆养殖中的应用探讨.《水产科技情报》.2007,第34卷(第6期),第271－274页. * |
| 李秋芬 等.复合有益菌剂对工厂化大菱鲆育苗水净化效果研究.《水产学报》.2006,第30卷(第6期),第852-856页. * |
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