CN102283328A - Multifunctional compound micro-ecological preparation for culturing holothurian - Google Patents
Multifunctional compound micro-ecological preparation for culturing holothurian Download PDFInfo
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Abstract
The invention discloses a multifunctional compound micro-ecological preparation for culturing holothurian, which comprises Bacillus atrophaeus, Bacillus subtilis, Bacillus cereus and a carrier. The symbiosis among probiotic bacteria screened by the multifunctional compound microecological preparation for culturing the holothurian is good; the multifunctional compound micro-ecological preparation for culturing the holothurian has a better antagonistic action on the pathogenic bacteria of the holothurian, has the functions of increasing the growth speed, the immunity and the vitality of the digestive enzyme of the intestinal tract of the holothurian and achieves multiple effects of antibiosis, immunity and growth promotion through the compatibility of the probiotic bacteria. Indigenous floras screened from the intestinal tract of the holothurian is safe and nontoxic for the holothurian, has a high breeding speed, is easy to culture, has high environment adaptation and can be faster bred to form predominant bacteria. The application method is simple and convenient, i.e. only the preparation needs to be added to a feed in proportion to be thrown to feed the holothurian. The multifunctional compound microecological preparation for culturing the holothurian is suitable for breeding and culturing young holothurians and controlling the diseases of the holothurian under various culture modes. The multifunctional compound micro-ecological preparation for culturing the holothurian plays an important role in improving the quality of holothurian merchandises and ensuring the food safety.
Description
Technical field
The present invention relates to the microorganism formulation technical field, particularly a kind ofly comprise three kinds of probios, have the Mobyneb holothruian cultures compound micro-ecological preparation of multi-efficiencies such as antibiotic, immune, growth promotion.
Background technology
Beneficial bacteria of intestinal tract is that a kind of alimentary canal that derives from mostly acts on gastral active bacteria formulation again, can effectively replenish the beneficial microbe in the animal alimentary canal, suppress growth of pathogenic bacteria, improve the archenteric flora balance, improve the immunity function or the speed of growth of aquatic livestock.
Stichopus japonicus (Apostichopus japonicus Liao claims imitative stichopus japonicus again) belongs to invertebrate, Echinodermata, and Holothuroidea, imitative stichopus japonicus belongs to.Because of it is of high nutritive value, the breed scale constantly enlarges, and has developed into one of sea-farming kind of the coastal maximum of China at present.In recent years because the relatively lagging behind of stichopus japonicus high-density breeding and running lack of standardization and disease control technology, disease problem order becomes outstanding, especially culture breaking out of stichopus japonicus " Beancurd sheet syndrome ", caused serious economy loss, restricted the healthy and sustainable development of this industry.Because special life habit and the breed characteristics of stichopus japonicus, and prophylaxis such as antibiotic medicine itself exists some shortcomings and limitation again, prevents one of effective measures of apostichopus japonicus culture disease generation so adopt probiotics to become.
According to deriving from nature, the principle of back to nature, screening is the preferred approach of exploitation probiotics with the bacterial classification that former host, former habitat, former little ecosystem, pathogenic microorganism groupy phase adapt to.In the face of the situation of the special-purpose probiotics scarcity of present sea cucumber, screen probio and be developed to probiotics from the sea cucumber enteron aisle, avoided the difference between the different strains, guarantee the security and the high efficiency of probiotics.Therefore, from the sea cucumber enteron aisle, separate the indigenous probiotic bacterium, reach antibiotic, immune, somatotrophic multiple effect, the sea cucumber healthy aquaculture is had realistic meaning by compatibility to relevant bacterial strain.
March toward 21 century, holothruian cultures already develop rapidly, 84200 hectares of cultured areas, and 20,000,000,000 yuan of year wound output values have produced huge economic and social benefit, become one of pillar industry of northern China aquaculture.Along with holothruian cultures already develop rapidly, the aquaculture model of high density, batch production, intensification makes the ecological environment imbalance serious, and various diseases come one after another.In recent years, sea cucumber various bacterial diseases such as " Beancurd sheet syndromes " popular makes the disease problem become the main bottleneck of this industry development.The application of chemicalses such as antibiotic is bred, is prevented that the sea cucumber disease from having played certain positive role the control growth of pathogenic bacteria, but owing to use chemicals such as antibiotic for a long time, in a large number, pathogen also produces drug resistance, flora imbalance in sea cucumber enteron aisle and the breeding environment, the sea cucumber immunity degradation, the various diseases recurrence rate is higher, and medicament residue is serious simultaneously, is endangering cultivated animals body and human health.Therefore, the Application and Development probiotics has important practical significance to holothruian cultures.The life of dwelling at the end of sea cucumber battalion, can play by the enteral microecological formulation that adds in the feed of ingesting suppress harmful bacterium, improve premunition, effect such as promotion growth.
Probio derives from alimentary canal mostly and acts on gastral active bacteria formulation again, can effectively replenish the beneficial microbe in the animal alimentary canal, suppress the pathogen growth, improve the archenteric flora balance, improve the immunity function or the speed of growth of aquatic livestock, therefore, probiotics is a feed addictive of using and study more green, safety at present always.In recent years, people begin to utilize probiotics to change little ecological composition in cultivated animals enteron aisle and the environment, thereby improve the immunity of breeding ecological environment, raising cultivated animals, especially the development and application of the compound micro-ecological preparation of being made up of many bacterial classifications, the direction of culturing towards green health for culture fishery develop and play positive role.
More about the report of aquatic livestock probio before the present invention, as: the quick grade of power " prebiotics for aquaculture " (number of patent application: 200610042574.0), (number of patent application: 200710057106.5), and the report that relevant sea cucumber enteron aisle and breeding environment probio are screened is less for Wang Lianmin etc. " promote fresh-water fishes grow probiotic composition ".The selection of bacterial classification is the key link of little ecological goods in the probiotics.Selecting the standard of bacterial classification must be guidance with the microecology theory, and according to deriving from nature, the principle of back to nature is selected with the bacterial classification of former host, former habitat, former little ecosystem, the adaptation of pathogenic microorganism groupy phase the most desirable.The probio that comes from animal itself has avoided the difference between the different strains, and having guaranteed has higher specific aim to the host, has improved the security and the high efficiency that are applied to the host.Therefore, from the sea cucumber enteron aisle, separate the indigenous probiotic bacterium, reach antibiotic, immune, somatotrophic multiple effect, the sea cucumber healthy aquaculture is had realistic meaning by compatibility to relevant bacterial strain.
Summary of the invention
Technical problem to be solved by this invention is, a kind of multi-functional compound micro-ecological preparation that adapts to antibiotic, immune, the growth promotion multi-efficiency of sea cucumber enteron aisle and breeding environment is provided, thereby reduces the use of chemicalses such as antibiotic in the holothruian cultures process.It is good that the present invention obtains the probio security, the sea cucumber pathogen is suppressed effective, and can improve the death rate of sea cucumber specific growth rate, reduction sea cucumber, immunity and the enteron aisle digestive enzyme activity of raising sea cucumber, reach antibiotic, immune, somatotrophic multiple effect.
For solving the problems of the technologies described above, the invention provides a kind of Mobyneb holothruian cultures compound micro-ecological preparation, it is characterized in that, comprise: atrophy bacillus atrophaeus, bacillus subtilis Bacillus subtilis, bacillus cereus Bacillus cereus, and carrier.
Described atrophy bacillus is preferably: be located away from wild stichopus japonicus enteron aisle, be preserved in Chinese typical culture collection center, preserving number is CCTCC M 2010316.
Described bacillus subtilis is preferably: be located away from wild stichopus japonicus enteron aisle, be preserved in Chinese typical culture collection center, preserving number is CCTCC M 2010314.
Described bacillus cereus is preferably: be located away from wild stichopus japonicus enteron aisle, be preserved in Chinese typical culture collection center, preserving number is CCTCC M 2010315.
Described carrier is preferably: particle diameter≤80 purpose zeolite powders.
In the described compound micro-ecological preparation, atrophy bacillus atrophaeus, bacillus subtilis Bacillus subtilis, bacillus cereus Bacillus cereus, ratio of weight and number be preferably: 1～2 part: 1～2 part: 1～2 part.
For solving the problems of the technologies described above, the present invention also provides a kind of bacillus subtilis strain, and described bacterial strain is located away from wild stichopus japonicus enteron aisle, is preserved in Chinese typical culture collection center, and preserving number is CCTCCM 2010314.
For solving the problems of the technologies described above, the present invention provides a kind of feed addictive again, comprising: bacillus subtilis strain, described bacterial strain are located away from wild stichopus japonicus enteron aisle, are preserved in Chinese typical culture collection center, and preserving number is CCTCC M 2010314.
For solving the problems of the technologies described above, the present invention provides the application of a kind of described bacillus subtilis strain as feed addictive, microorganism formulation and/or enteron aisle prebiotics in addition.
For solving the problems of the technologies described above, the present invention provides the application of a kind of described bacillus subtilis strain in the preparation of feed addictive, microorganism formulation and/or enteron aisle prebiotics again.
The present invention has following beneficial technical effects:,
1) symbiosis is good between the screening probio, the sea cucumber pathogen there is antagonism preferably, and have the function that improves the sea cucumber speed of growth, immunity and enteron aisle digestive enzyme activity, reached antibiotic, immune, somatotrophic multi-efficiency by the compatibility between the probio.
2) the screening probio is the indigenous bacterium of screening from the sea cucumber enteron aisle, fast to sea cucumber safety non-toxic, reproduction speed, be easy to cultivate, strong to adaptive capacity to environment, can breed the formation dominant bacteria faster.
3) using method is easy, only said preparation need be added to throw something and feed in the feed to getting final product according to proportioning, is applicable to the sea cucumber disease control of children's ginseng cultivating and various aquaculture models.
4) use the present invention to reduce or substitute antibiotic use, to reduce medicine residual, improve the sea cucumber commercial quality, ensuring food safety has important function.
Description of drawings
Fig. 1 is influence the as a result figure of the described probiotics of the embodiment of the invention to sea cucumber coelomic fluid acid phosphatase enzyme activity;
Fig. 2 is influence the as a result figure of the described probiotics of the embodiment of the invention to sea cucumber coelomic fluid alkaline phosphatase activity;
Fig. 3 is influence the as a result figure of the described probiotics of the embodiment of the invention to sea cucumber enteron aisle proteinase activity;
Fig. 4 is influence the as a result figure of the described probiotics of the embodiment of the invention to sea cucumber enteron aisle amylase activity.
The specific embodiment
Describe embodiments of the present invention in detail below with reference to drawings and Examples, how the application technology means solve technical problem to the present invention whereby, and the implementation procedure of reaching technique effect can fully understand and implements according to this.
Embodiment 1:
Screening of potential probio and safety experiment
Get wild healthy sea cucumber, under aseptic condition, sea cucumber is carried out vivisection, get its enteron aisle, extrude intestinal contents, with 15 ‰ normal saline flushing enteron aisle of sterilization three times, the enteron aisle homogenate with after the flushing is coated on after the proportional diluted on TSB culture medium and the 2216E seawater agar medium, select suitable dilution flat board to carry out the separation of bacterium, obtain 240 strain bacteriums altogether.Bacterium to 240 strain pure cultures is measured its product enzyme activity with casein hydrolyzing culture medium and the cultivation of starch hydrolyzing culture medium, be indicator bacteria with sea cucumber encountered pathogenic bacteria-Vibrio splindidus (Vibrio splendidus), Pseudoalteromonas (Pseudoalteromonas nigrifaciens) and Aeromonas hydrophila (Aeromonas hydrophlia) simultaneously, utilize dibbling method, right-angled intersection method and Oxford agar diffusion method to detect the bacteriostasis of institute's separation of bacterial, filtered out 3 strain good antimicrobial effect and the strong bacterium of enzymatic productivity altogether the sea cucumber pathogen.By this 3 strain bacterium is carried out form, Physiology and biochemistry and Molecular Identification, the result shows that 3 probiotics that screening obtains are respectively atrophy bacillus (Bacillus atrophaeus), bacillus subtilis (Bacillus subtilis), bacillus cereus (Bacillus cereus), this 3 strain bacterium is preserved in Chinese typical culture collection center (being called for short CCTCC), and preserving number is respectively: atrophy bacillus CCTCC M 2010316, bacillus subtilis CCTCC M 2010314, bacillus cereus CCTCC M 2010315.Antagonistic experiment result between the 3 strain bacterium shows between the 3 strain bacterium does not all have antagonism, and geometric ratio is carried out compatibility, can significantly improve the antagonism to pathogenic bacteria, the results are shown in Table 1.
Table 1: compatibility bacterial strain and single strain antagonism comparing result
The 3 strains bacterial strain to be detected of picking activation 18h makes 1 * 10 with the physiological saline of 15 ‰ sterilizations respectively
10The bacteria suspension of CFU/mL, proportional diluted to 1 * 10
5CFU/mL.Method with injection is carried out infection experiment, every sea cucumber injection 0.2mL bacteria suspension, negative control is made as the sterile saline group of injection same dose, the group of blank group for it not being carried out any processing, positive control is made as the Pseudoalteromonas (P.nigrifaciens) of the variable concentrations gradient of injection same dose, after the infection, feeding activity, physiological reaction, the body surface of observing sea cucumber every day change and death condition.The result shows: death in various degree appears in the positive control sea cucumber, and death condition does not all appear in 3 probiotics experimental group, just 1 * 10
10CFU/mL injection group occurs ingesting differing from indivedual sea cucumbers and tells dirty phenomenon, 1 * 10
9Bad reaction does not all appear in the following group of CFU/mL.Confirm by above-mentioned test: 3 strain bacterium are to the sea cucumber safety non-toxic.
Embodiment 2:
The preparation of three kinds of probiotics active preparations
The preparation method of atrophy bacillus (Bacillus atrophaeus) CCTCC M 2010316 bacterium powder, bacillus subtilis (Bacillus subtilis) CCTCC M 2010314 bacterium powder, bacillus cereus (Bacilluscereus) CCTCC M 2010315 bacterium powder comprises the steps:
A, dull and stereotyped cultivation rejuvenation: atrophy bacillus, bacillus subtilis, the bacillus cereus with-80 ℃ of preservations is inoculated on the TSB plating medium respectively, in 28 ℃ of cultivation 24h, makes rejuvenation of spawn and forms single bacterium colony;
The preparation of B, seed liquor: the bacterial classification that steps A is cultivated is made 1 * 10 with the physiological saline of 15 ‰ sterilizations
5The bacteria suspension of CFU/mL, the inoculum concentration with 5% are inoculated in the 250mL conical flask that fills the aseptic TSB fluid nutrient medium of 100mL, and 28 ℃, 170rpm shaken cultivation 18～20h;
C, fermented and cultured: the seed liquor among the step B is inoculated in 5% inoculum concentration fills 2L and do not have in the 5L triangular flask of bacteria fermentation culture medium, 28 ℃, 170rpm shaken cultivation 84h, to viable count be 2～4 * 10
9CFU/mL, stuck fermentation;
Described fermentation medium is a glucose 1～2%, soybean muffin 1～2%, and sodium chloride 1%, all the other are water;
The preparation of D, probiotic active microbial inoculum: in the zymotic fluid of step D, add zeolite powder, add the 10g zeolite powder in every 100mL zymotic fluid, static 40min behind the 170rpm vibration 40min, with above-mentioned zymotic fluid in 4 ℃ of centrifugal 20min of 6000rpm, abandon supernatant, after the zeolite powder that adds 20～40g in the every 100g precipitation again mixes, in 45～50 ℃ of oven dry 3～4h, obtain the probiotic active microbial inoculum of 3 strain bacterium respectively, the viable bacteria measurement result shows that prepared microbial inoculum viable bacteria content reaches 1 * 10
9More than the CFU/g.
Embodiment 3:
The multi-functional holothruian cultures preparation of compound micro-ecological preparation: with atrophy bacillus CCTCC M2010316 bacterium powder, bacillus subtilis CCTCC M 2010314 bacterium powder, bacillus cereus CCTCCM 2010315 bacterium powder, carry out compatibility at 1: 1: 1 according to weight proportion, mixing prepares compound micro-ecological preparation.
Embodiment 4:
Compound micro-ecological preparation is to the influence of growth, survival rate, immune indexes and the enteron aisle digestive enzyme activity of sea cucumber.
Prepared compound micro-ecological preparation is added in the feed to the sea cucumber of throwing something and feeding test according to 3 ‰ of feed weight.
Test is picked up from certain holothruian cultures company with sea cucumber, support after 7 days temporarily, select similar, the vigor of size preferably sea cucumber test, the sea cucumber body weight is initially 11.94 ± 2.38g, the long 4.96 ± 1.87cm of body, duration of test, seawater salinity is 29～31, and the pH value is 7.46～8.02, and dissolved oxygen is more than the 5.0g/L, water temperature is 12～16 ℃, and the test period is 60 days.Test is divided into 3 groups, control group is thrown something and fed and is not added the holothurian feed of probiotics, test group 2 is added the holothurian feed of 3 ‰ single bacillus subtilis formulations for throwing something and feeding, test group 3 is added the holothurian feed of 3 ‰ compound micro-ecological preparations of the present invention for throwing something and feeding, every group of 3 repetitions, each repeats 15 sea cucumbers.
0d, 30d and 60d in test measure the sea cucumber body weight and calculate specific growth rate (SGR) and survival rate, measure sea cucumber enteron aisle proteinase activity and amylase activity, measure acid phosphatase (ACP), alkaline phosphatase (AKP) activity of sea cucumber coelomic fluid.After the off-test of throwing something and feeding, from each repeating groups, randomly draw 10 sea cucumbers, manually attack malicious infection experiment with Vibrio splindidus, it is 1 * 10 that every sea cucumber is injected 200 μ L concentration
9The Vibrio splindidus bacterium liquid of CFU/mL, record are attacked three week of poison back interior two groups the death rate.
Result of the test is as follows:
Aspect growth promotion, utilizing single bacillus subtilis formulation and compound micro-ecological preparation of the present invention to adopt to mix to raise throws something and feeds, carry out the holothruian cultures test, result of the test is as shown in table 2, add single bacillus subtilis formulation and compound micro-ecological preparation group of the present invention, all can significantly improve the specific growth rate (SGR, P＜0.05) of sea cucumber, and can improve the survival rate of sea cucumber.
Table 2: probiotics is to the influence of sea cucumber survival rate and specific growth rate
As depicted in figs. 1 and 2, be respectively probiotics to sea cucumber coelomic fluid acid phosphatase enzyme activity influence as a result figure and, probiotics is to the figure as a result that influences of sea cucumber coelomic fluid alkaline phosphatase activity.Wherein, contain same letter difference not remarkable (P＞0.05); Contain different alphabetical significant differences (P＜0.05).
Aspect raising immunity, at duration of test, ACP, the AKP activity of adding compound micro-ecological preparation group of the present invention all are higher than other group; Add single bacillus group and do not have significant difference (P＞0.05) with control group.Result of the test is as depicted in figs. 1 and 2: add compound micro-ecological preparation in the feed and can significantly improve ACP in the sea cucumber coelomic fluid, AKP enzymatic activity.
As shown in Figure 3 and Figure 4, be respectively the described probiotics of the embodiment of the invention to sea cucumber enteron aisle proteinase activity influence as a result figure and, the described probiotics of the embodiment of the invention is to the figure as a result that influences of sea cucumber enteron aisle amylase activity.Wherein, contain same letter difference not remarkable (P＞0.05); Contain different alphabetical significant differences (P＜0.05).
Aspect raising enteron aisle digestive enzyme activity, at duration of test, adding compound micro-ecological preparation histone enzyme of the present invention and amylase activity is significantly higher than blank group and adds single probiotics group, add single probiotics group digestive enzyme activity and be higher than the blank group, but significantly be lower than compound microecological interpolation group.Result of the test is as shown in Figure 3 and Figure 4: add the digestive enzyme activity that compound micro-ecological preparation can significantly improve the stichopus japonicus enteron aisle in the feed.
Aspect raising sea cucumber premunition; after manually attacking poison; the death rate of single bacillus subtilis formulation and compound micro-ecological preparation sea cucumber of the present invention significantly is lower than control group (P＜0.05); wherein; the death rate of adding compound micro-ecological preparation group sea cucumber of the present invention minimum (P＜0.05); with single bacillus subtilis group significant difference; result of the test is as shown in table 3; showing has certain synergy between 3 probiotics, compound micro-ecological preparation reaches 81.25% to the immune protective rate of stichopus japonicus.
Table 3: the immune protective rate of the death rate of stichopus japonicus and probiotics on the same group not in the artificial challenge experiment
Raise the holothruian cultures result of the test of throwing something and feeding and prove by mixing: multi-functional compound micro-ecological preparation of the present invention with 3 ‰ addition (the 1kg feed adds this probiotics of 3g) as feed addictive, the sea cucumber specific growth rate be can improve, the sea cucumber death rate, the immunocompetence that strengthens sea cucumber and the digestive enzyme activity of enteron aisle reduced, improve the premunition of sea cucumber simultaneously, reached antibiotic, immune, growth promotion multiple effect, can reduce and replace antibiotic use, culture survival rate and product quality to improving, ensure that the stable and high yields of this industry has crucial meaning.
Embodiment 5:
Mobyneb holothruian cultures probiotics, said preparation comprise a kind of carrier and three kinds of probios.
Wherein, described probiotics strain comprises atrophy bacillus (Bacillus atrophaeus), bacillus subtilis (Bacillus subtilis), bacillus cereus (Bacillus cereus), these bacterial strains are stronger to the pathogen antagonism of sea cucumber, have higher product protease and diastatic ability, and do not have antagonism between 3 probiotics.Described 3 probiotics separate, screen gained by the applicant from wild healthy sea cucumber enteron aisle, identify through form, Physiology and biochemistry and dna molecular, and being preserved in Chinese typical culture collection center (be called for short CCTCC) on November 25th, 2010, preserving number is respectively: atrophy bacillus CCTCC M 2010316, bacillus subtilis CCTCC M 2010314, bacillus cereus CCTCC M 2010315.
Probiotics of the present invention also comprises carrier, carrier is≤80 purpose zeolite powders, with the zeolite powder is preparing carriers atrophy bacillus CCTCC M 2010316 bacterium powder, bacillus subtilis CCTCC M2010314 bacterium powder, bacillus cereus CCTCC M 2010315 bacterium powder, and prepared bacillus preparation viable bacteria content all reaches 1 * 10
9More than the CFU/g.
Probiotics of the present invention comprises atrophy bacillus CCTCC M 2010316 bacterium powder, bacillus subtilis CCTCC M 2010314 bacterium powder, bacillus cereus CCTCC M 2010315 bacterium powder, and the weight proportion of above-mentioned each composition is 1: 1: 1.
Probiotics of the present invention is a kind of green feed additive, and addition is advisable according to 3 ‰ of feed gross weight.
Embodiment 6:
Mobyneb holothruian cultures compound micro-ecological preparation.
Comprise: atrophy bacillus atrophaeus, bacillus subtilis Bacillussubtilis, bacillus cereus Bacillus cereus, and carrier.
Described atrophy bacillus is preferably: be located away from wild stichopus japonicus enteron aisle, be preserved in Chinese typical culture collection center, preserving number is CCTCC M 2010316.
Described bacillus subtilis is preferably: be located away from wild stichopus japonicus enteron aisle, be preserved in Chinese typical culture collection center, preserving number is CCTCC M 2010314.
Described bacillus cereus is preferably: be located away from wild stichopus japonicus enteron aisle, be preserved in Chinese typical culture collection center, preserving number is CCTCC M 2010315.
Described carrier is preferably: particle diameter≤80 purpose zeolite powders.
In the described compound micro-ecological preparation, atrophy bacillus atrophaeus, bacillus subtilis Bacillus subtilis, bacillus cereus Bacillus cereus, ratio of weight and number be preferably: 1～2 part: 1～2 part: 1～2 part.
Embodiment 7:
Bacillus subtilis strain.
Described bacterial strain is located away from wild stichopus japonicus enteron aisle, is preserved in Chinese typical culture collection center, and preserving number is CCTCC M 2010314.
Embodiment 8: feed addictive.
Comprise: bacillus subtilis strain, described bacterial strain are located away from wild stichopus japonicus enteron aisle, are preserved in Chinese typical culture collection center, and preserving number is CCTCC M 2010314.
Embodiment 9: bacillus subtilis strain is as the application of feed addictive, microorganism formulation and/or enteron aisle prebiotics.
Embodiment 10: the application of bacillus subtilis strain in the preparation of feed addictive, microorganism formulation and/or enteron aisle prebiotics.
This intellectual property of primary enforcement that all are above-mentioned is not set restriction this new product of other forms of enforcement and/or new method.Those skilled in the art will utilize this important information, and foregoing is revised, to realize similar implementation status.But all modifications or transformation belong to the right of reservation based on new product of the present invention.
The above only is preferred embodiment of the present invention, is not to be the restriction of invention being made other form, and any those skilled in the art may utilize the technology contents of above-mentioned announcement to be changed or be modified as the equivalent embodiment of equivalent variations.But every technical solution of the present invention content that do not break away to any simple modification, equivalent variations and remodeling that above embodiment did, still belongs to the protection domain of technical solution of the present invention according to technical spirit of the present invention.
Claims (10)
1. a Mobyneb holothruian cultures compound micro-ecological preparation is characterized in that, comprising: atrophy bacillus atrophaeus, bacillus subtilis Bacillus subtilis, bacillus cereus Bacillus cereus, and carrier.
2. according to the described Mobyneb holothruian cultures of claim 1 compound micro-ecological preparation, it is characterized in that described atrophy bacillus is located away from wild stichopus japonicus enteron aisle, be preserved in Chinese typical culture collection center, preserving number is CCTCC M 2010316.
3. according to claim 1 or 2 described Mobyneb holothruian cultures compound micro-ecological preparations, it is characterized in that described bacillus subtilis is located away from wild stichopus japonicus enteron aisle, be preserved in Chinese typical culture collection center, preserving number is CCTCC M 2010314.
4. according to each described Mobyneb holothruian cultures compound micro-ecological preparation in the claim 1～3, it is characterized in that described bacillus cereus is located away from wild stichopus japonicus enteron aisle, be preserved in Chinese typical culture collection center, preserving number is CCTCC M 2010315.
5. according to each described Mobyneb holothruian cultures compound micro-ecological preparation in the claim 1～4, it is characterized in that described carrier is particle diameter≤80 purpose zeolite powders.
6. according to each described Mobyneb holothruian cultures compound micro-ecological preparation in the claim 1～5, it is characterized in that, in the described compound micro-ecological preparation, atrophy bacillus atrophaeus, bacillus subtilis Bacillus subtilis, bacillus cereus Bacillus cereus, ratio of weight and number be: 1～2 part: 1～2 part: 1～2 part.
7. a bacillus subtilis strain is characterized in that, described bacterial strain is located away from wild stichopus japonicus enteron aisle, is preserved in Chinese typical culture collection center, and preserving number is CCTCC M 2010314.
8. a feed addictive is characterized in that, comprising: bacillus subtilis strain, described bacterial strain are located away from wild stichopus japonicus enteron aisle, are preserved in Chinese typical culture collection center, and preserving number is CCTCCM 2010314.
9. one kind as the application of bacillus subtilis strain as described in the claim 7 as feed addictive, microorganism formulation and/or enteron aisle prebiotics.
10. one kind as the application of bacillus subtilis strain in the preparation of feed addictive, microorganism formulation and/or enteron aisle prebiotics as described in the claim 7.
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| CN106578731A (en) * | 2016-12-20 | 2017-04-26 | 大连赛姆生物工程技术有限公司 | Marine synbiotics additive and application thereof in stichopus japonicus culture |
| CN107937300A (en) * | 2017-11-08 | 2018-04-20 | 青岛农业大学 | A kind of bacillus subtilis and its application in aquaculture |
| CN111789054A (en) * | 2020-07-29 | 2020-10-20 | 辽宁省海洋水产科学研究院 | Method for applying bacillus liquid to stichopus japonicus seedling culture |
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| CN102524604B (en) * | 2012-03-05 | 2013-04-17 | 王贵国 | Biological feeds for sea cucumber |
| CN102524604A (en) * | 2012-03-05 | 2012-07-04 | 王贵国 | Biological feeds for sea cucumber |
| CN102652536A (en) * | 2012-04-06 | 2012-09-05 | 山东省海水养殖研究所 | Formula and processing technology of stichopus japonicus compound micro-ecological nutritional additive |
| CN102899264B (en) * | 2012-06-25 | 2016-09-21 | 石河子大学 | A kind of microbial-bacterial fertilizer and its production and use |
| CN102899264A (en) * | 2012-06-25 | 2013-01-30 | 石河子大学 | Microbial fertilizer and its preparation method and use |
| CN103845367A (en) * | 2014-01-03 | 2014-06-11 | 青岛东海药业有限公司 | Micro-ecological preparation applied to aquaculture of abalones and sea cucumbers |
| CN105746947A (en) * | 2016-03-07 | 2016-07-13 | 五河县金满塘生态种养殖家庭农场 | Microecological preparation for improving intestinal function of Elopichthys bambusa |
| CN106417924A (en) * | 2016-09-05 | 2017-02-22 | 河北省海洋与水产科学研究院 | Method for preparing trepang feed added scallop net cage attachments |
| CN106417924B (en) * | 2016-09-05 | 2019-08-16 | 河北省海洋与水产科学研究院 | Add the preparation method of the holothurian feed of scallop cylinder mould attachment |
| CN106578731A (en) * | 2016-12-20 | 2017-04-26 | 大连赛姆生物工程技术有限公司 | Marine synbiotics additive and application thereof in stichopus japonicus culture |
| CN107937300A (en) * | 2017-11-08 | 2018-04-20 | 青岛农业大学 | A kind of bacillus subtilis and its application in aquaculture |
| CN107937300B (en) * | 2017-11-08 | 2020-09-01 | 青岛农业大学 | Bacillus subtilis and application thereof in aquaculture |
| CN111789054A (en) * | 2020-07-29 | 2020-10-20 | 辽宁省海洋水产科学研究院 | Method for applying bacillus liquid to stichopus japonicus seedling culture |
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