CN109321505A - A kind of complex microorganism preparations adjusting aquatic livestock enteron aisle - Google Patents
A kind of complex microorganism preparations adjusting aquatic livestock enteron aisle Download PDFInfo
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Abstract
The invention discloses a kind of complex microorganism preparations for adjusting aquatic livestock enteron aisle, the complex microorganism preparations are made of Lactobacillus acidophilus, bacillus coagulans, bacillus licheniformis, bacillus subtilis, bacillus amyloliquefaciens, bacterium number quantity ratio is 1.8-2.2: 0.8-1: 1.8-2.2: 1.8-2.2: 0.8-1, and total bacteria count reaches 10,000,000,000/g.Complex microorganism preparations of the present invention have the characteristics that nontoxic, safe and efficient, without secondary pollution, can not only prevent and treat aquatic livestock intestines problem, help digest absorption and improve immunity, and can improve water quality.
Description
Technical field
The invention belongs to technical field of aquaculture, more particularly, to a kind of complex microorganism for adjusting aquatic livestock enteron aisle
Preparation.
Background technique
20th century, the discovery of antibiotic promote the huge advance of physianthropy, and antibiotic becomes what antibacterial was cured the disease since then
Choice drug.A large amount of antibiotic is widely used, and so that animal farming industry is constantly benefited, especially culture fishery.In drug
On the basis of constantly being reformed with technology, high density, intensive, scale, batch production aquaculture model reach its maturity, aquatic livestock
Aquaculture was once advanced with swift and violent speed.But following disease is largely propagated, antibiotic is abused wantonly, cultivates ring
The problems such as border deteriorates, cause of disease drug resistance is reinforced, cultivated animals medicament residue also has become people's hot spot of interest.
Current culture fishery requires to introduce advanced biotechnology, microorganism formulation in quite wide field
Application then be one kind to the above-mentioned highly effective solution route of culture fishery Problems Existing.Some microorganism formulations enter water
After body, the harmful substances such as ammonia nitrogen, the hydrogen sulfide in water can be eliminated rapidly, improve the water environment of breeding water body, balancing alkalinity acidity,
The organic matters such as excreta, residual bait, the planktonic organism residuum of cultivated animals are decomposed into rapidly carbon dioxide, nitrate, phosphate etc.,
Nutrition is provided, promotes the breeding of unit cell algae, while itself breeding rapidly and forming dominant bacteria, to inhibit the life of pathogenic microorganism
It is long.The mechanism of action of microorganism formulation is other than direct bacteriostasis, and there are also other effects, that is, to immune system
Influence.It prompts the gross differences of microorganism formulation and antibiotic, and becomes an important reason of substitute antibiotics.One
Serial experiment shows that the immunity of animal itself, immune indexes can be enhanced in the presence of the intracorporal beneficial flora of aquatic livestock
There is close contact with beneficial flora.Many scholars have begun the research of the project, and achieve certain achievement.Warm Chongqing
Etc. having filtered out two plants of prebiotic bacillus of prawn；Lin Yu has studied A40209CDC4 (bacillus subtilis Bacillu
Ubtilis) and A31009NA (less move Sphingol single-cell Sphingomonas paucimobil) is to water quality and feed addition
Probiotic effects；Li Zhuojia etc. has studied Gracilaria Tenuistipitata V. Liui Gracilarla tenuistipitata var.1iui and benefit
Raw bacterium (photosynthetic bacteria Rhodopseudomoans ca psulata and bacillus lichenifirmis) is for 24 hours
The interior synergistic purification to intensive prawn culturing waste water acts on；The researchs such as Hua Xueming find that polysaccharide and probiotics are low to immune function
Under fugu obscurus there is immunoregulation effect, and the digestion of the young can be improved, improve growth efficiency；Wang Fu is strong etc. to be ground
Proteinase-producing bacteria has been studied carefully to Growth of The Juvenile Flounder Olivaceus facilitation；Yuan Chengyu etc. has studied probiotics and grows to young stichopus japonicus
And the influence of digestive enzyme activity.
The research of microorganism formulation develops for the direction of culture fishery towards green and healthy cultivation provides theoretical foundation,
For solve in current aquaculture disease is largely propagated, antibiotic abuse wantonly, breeding environment deterioration, the reinforcement of cause of disease drug resistance,
The problems such as cultivated animals medicament residue, has established certain basis.Therefore, develop it is a kind of adjust aquatic livestock enteron aisle it is compound micro-
Biological agent is particularly important.But there is also some defects for current microorganism formulation: safety is poor, and effect is unstable, strain
Source is indefinite (such as commercially available), and use condition is limited, although being effectively unable to improve water quality etc. to breeding water body disease.
Summary of the invention
The invention solves first technical problem be to provide it is a kind of adjust aquatic livestock enteron aisle complex microorganism system
Agent.The complex microorganism preparations have the characteristics that nontoxic, safe and efficient, without secondary pollution, can not only prevent and treat aquatic livestock enteron aisle
Disease helps digest absorption and improves immunity, and can improve water quality.
The invention solves second technical problem be to provide it is a kind of adjust aquatic livestock enteron aisle complex microorganism system
The preparation method of agent.
In order to solve the above technical problems, the present invention adopts the following technical scheme that:
A kind of complex microorganism preparations adjusting aquatic livestock enteron aisle, by Lactobacillus acidophilus, bacillus coagulans, lichens
Bacillus, bacillus subtilis, bacillus amyloliquefaciens composition, bacterium number quantity ratio is 1.8-2.2: 0.8-1: 1.8-2.2:
1.8-2.2: 0.8-1, total bacteria count reaches 10,000,000,000/g.
As the further improvement of technical solution, the Lactobacillus acidophilus, bacillus coagulans, bacillus licheniformis,
Bacillus subtilis, bacillus amyloliquefaciens bacterium number quantity ratio be 2: 1: 2: 2: 1.
As the further improvement of technical solution, additive amount of the complex microorganism preparations in breeding water body is 3-
8ppm, the additive amount in feed are 0.5-1%.
The complex microorganism preparations are used to improve the water quality of breeding water body, and it is flat to adjust the intracorporal Tiny ecosystem of aquatic livestock
Weighing apparatus improves immunity, promotes growth, to play the role of disease preventing and treating and salubrious.
To solve above-mentioned second technical problem, a kind of above-mentioned complex microorganism system for adjusting aquatic livestock enteron aisle of the present invention
The preparation method of agent, includes the following steps:
S1, screened from grass carp body, dissect grass carp, take out entire enteron aisle, with 3-5 DEG C of sterile saline repeated flushing,
It is put into homogenizer, is homogenized, takes 1.0g sample, be put into equipped with 9mL sterile water and be placed in the 50mL triangular flask of bead, set and shake
18-22min is vibrated on bed disperses microbial cell, stands 25-35 seconds up to 10-1Dilution；
S2,10 are drawn with 1mL aseptic straw-1Dilution 1mL is moved into the test tube equipped with 9mL sterile water, and suction is blown 3 times, is mixed
Combined bacteria liquid, 10-2Dilution；And so on, serial dilution is made 10-3、10-4、10-5、10-6、10-7、10-8It is a series of
Dilute bacterium solution；
S3, from 10-6、10-7、10-8It respectively takes 1mL in plate in dilution bacterium solution, pours into respectively and melt and be cooled to 45-50
DEG C nutrient broth agar culture medium and MRS culture medium, gently pivotal plate, is uniformly mixed bacterium solution with culture medium；After cooling
It is inverted, is observed after culture 24-48 hours in 28-30 DEG C；The bacterial strain separated from solid plate is if not single bacterium breeding
It forms, then further progress isolates and purifies；Well-grown representational colony inoculation inclined-plane is selected on plate, is done simultaneously
Plate coating checking, if discovery is impure, picking colony is further separated, until obtaining pure culture body；
S4, isolated dominant bacteria is identified according to the morphological feature and physiological and biochemical property of bacterium；From the bacterium of bacterium
Fall form and bio-chemical characteristics determined dominant bacteria be Lactobacillus acidophilus, it is bacillus coagulans, bacillus licheniformis, withered
After careless bacillus, bacillus amyloliquefaciens, this 5 kinds of bacterium are subjected to single strain fermentation, then Lactobacillus acidophilus, condensation bud
Spore bacillus, bacillus licheniformis, bacillus subtilis, bacillus amyloliquefaciens mix in proportion, and bacterium number quantity ratio is 1.8-2.2
: 0.8-1: 1.8-2.2: 1.8-2.2: 0.8-1, total bacteria count reaches 10,000,000,000/g.
As the further improvement of technical solution, in step S4, the Lactobacillus acidophilus, bacillus coagulans, lichens
Bacillus, bacillus subtilis, bacillus amyloliquefaciens bacterium number quantity ratio be 2: 1: 2: 2: 1.
Unless otherwise specified, each raw material in the present invention can be obtained by commercially available purchase, equipment used in the present invention
The conventional equipment in fields can be used or carried out referring to the prior art of fields.
Compared with prior art, the invention has the following beneficial effects:
The present invention targetedly screens strain, using Lactobacillus acidophilus, bacillus coagulans, lichens gemma
Bacillus, bacillus subtilis, bacillus amyloliquefaciens cooperation is made, they compound together play microorganism synergistic effect and
Synergistic function can prevent and treat aquatic livestock intestines problem, helps digest absorption and improve immunity.
Complex microorganism preparations of the invention are without any side effects, while low in cost, moreover it is possible to have complicated
Machine object is converted into for simple inorganic matter needed for planktonic organism growth and breeding, maintains the ecological balance of culture environment of aquatic products, thus
Improve water quality.
Complex microorganism preparations of the invention pass through the compounding of type and ratio between multiple strains, and treatment effect is stablized,
And the limitation of no use condition, strain easily obtain, can satisfy the demand of existing breeding water body.
Specific embodiment
In order to illustrate more clearly of the present invention, below with reference to preferred embodiment, the present invention is described further.
A kind of complex microorganism preparations for adjusting aquatic livestock enteron aisle of the present invention, by Lactobacillus acidophilus, condensation gemma bar
Bacterium, bacillus licheniformis, bacillus subtilis, bacillus amyloliquefaciens composition, bacterium number quantity ratio is 1.8-2.2: 0.8-1:
1.8-2.2: 1.8-2.2: 0.8-1, total bacteria count reaches 10,000,000,000/g.
Lactobacillus acidophilus (Lactobacillus acidophilus), also known as A bacterium, are one kind of lactobacillus.A bacterium
It is suitble to existence in acidity, 30 DEG C of environment, there are the mankind, the intestines and stomach of animal, oral cavity and vaginas etc.；With most cream
Sour bacterium is the same to be changed into lactic acid for lactose.Its relevant strain can also generate ethyl alcohol, carbon dioxide, acetic acid.A bacterium can not be
Temperature and humidity is excessively high, environment of direct sunlight is survived.The Lactobacillus acidophilus can by purchase obtain, can also by from
It screens and obtains in aquatic livestock enteron aisle.
Bacillus is Gram-positive, is generally existing a kind of aerobic bacteria, can be formed at later stages
The bacillus of gemma (endospore).Bacillus can remove remains on Cultivated water bottom waste, especially Lao Chi for a long time
Abundant residues bait, discharged waste, plant and animal residues and the pernicious gas (ammonia nitrogen, hydrogen sulfide etc.) of bottom accumulation, are allowed to first
Be decomposed into small molecule (polypeptide, higher fatty acids etc.), after be decomposed into more small organic molecule (amino acid, lower fatty acid, list
Sugar, cyclic hydrocarbon etc.), it is finally decomposed to carbon dioxide, nitrate, sulfate etc., COD, BOD in water is effectively reduced, makes water body
In ammonia nitrogen, nitrite, sulfide concentration reduce, to effectively improve water quality.It simultaneously can also be to be with unicellular alga
Main phytoplankton provides nutriment, promotes breeding.The photosynthesis of these phytoplanktons, and exhaling for animal is produced for pool inner water
It inhales, the decomposition of organic matter offer oxygen, to form a benign ecological circulation.Due to bacillus mass propagation, in pond
Dominant population is formed, the breeding of pathogenic microorganism is can inhibit, reduces disease.Further, since the special construction of gemma makes bud
Spore bacillus is very strong to the resistance of the poor environments such as drying, high temperature and pressure, oxidation, strong acid and strong base, extruding, and stability is high, and
With very strong protease, lipase, amylase activity, there are a variety of effective enzymatic effects in intestinal colonisation, enteron aisle can be made
Interior pH value decline, NH3Concentration reduces, and starch, cellulose etc. is promoted to decompose.It is the bacillus coagulans, bacillus licheniformis, withered
Careless bacillus, bacillus amyloliquefaciens belong to bacillus, can be obtained by purchase, can also be by from aquatic livestock enteron aisle
Middle screening obtains.
Applicant is through long term test the study found that combining single culture or the other different strain of unitary class, for feeding
The organic matter for growing water body decomposes and prevents and treats disease of digestive tract, improves immunity and help digest absorption aspects, tends not to reach
It is ideal to require.And mix above-mentioned five kinds of strains, it is compounded, is mutually promoted by strain inter-species, syntrophism is complete
Comprehensive decomposition of pairs of pollutant, and cultivated animals immunity and premunition are improved, the list of every quasi-microorganism can be given full play to
Strain effect and it is compound after synergistic effect and synergistic function, the pollutants such as organic matter, ammonia nitrogen, nitrite in water body are dropped
Solution conversion, achievees the purpose that purify water and repairs water body；Intracorporal microecological balance is adjusted to aquatic livestock, is improved immune
Power promotes growth, to play the role of disease preventing and treating and salubrious.In addition, 5 kinds of microorganisms are aquatic livestock enteron aisle
In superior microorganism, can quickly be bred in pond waters and enteron aisle, and play compound synergistic effect.
In the certain preferred embodiments of the present invention, the Lactobacillus acidophilus, bacillus coagulans, bacillus licheniformis,
Bacillus subtilis, bacillus amyloliquefaciens bacterium number quantity ratio be 2: 1: 2: 2: 1.
In some embodiments of the invention, additive amount of the complex microorganism preparations in breeding water body is 3-8ppm,
Additive amount in feed is 0.5-1%.
Above-mentioned complex microorganism preparations are particularly suitable for organic matter, ammonia nitrogen, nitrite in degrading cultivation water, improve
Aquatic livestock immunity and premunition promote growth.
Applicant in practice, it has been found that according to special ratios using Lactobacillus acidophilus of the invention, bacillus coagulans,
Bacillus licheniformis, bacillus subtilis, bacillus amyloliquefaciens strain combination compounded, for example 2: 1: 2: 2: 1, than list
A certain kind is solely used, certain one kind is used alone or certain several combined effect is used alone and to get well.
The preparation method for inventing a kind of above-mentioned complex microorganism preparations for adjusting aquatic livestock enteron aisle, includes the following steps:
S1, screened from grass carp body, dissect grass carp, take out entire enteron aisle, with 4 DEG C of sterile saline repeated flushing, put
Enter in homogenizer, is homogenized, takes 1.0g sample, be put into equipped with 9mL sterile water and be placed in the 50mL triangular flask of bead, set shaking table
Upper oscillation 20min disperses microbial cell, stand 30s 10-1Dilution；
S2,10 are drawn with 1mL aseptic straw-1Dilution 1mL is moved into the test tube equipped with 9mL sterile water, and suction is blown 3 times, is mixed
Combined bacteria liquid, 10-2Dilution；And so on, serial dilution is made 10-3、10-4、10-5、10-6、10-7、10-8It is a series of
Dilute bacterium solution；
S3, from 10-6、10-7、10-8It respectively takes 1mL in plate in dilution bacterium solution, pours into respectively and melt and be cooled to 45-50
DEG C nutrient broth agar culture medium and MRS culture medium, gently pivotal plate, is uniformly mixed bacterium solution with culture medium；After cooling
It is inverted, is observed after culture 24-48 hours in 28-30 DEG C；The bacterial strain separated from solid plate is if not single bacterium breeding
It forms, then further progress isolates and purifies；Well-grown representational colony inoculation inclined-plane is selected on plate, is done simultaneously
Plate coating checking, if discovery is impure, picking colony is further separated, until obtaining pure culture body；
S4, isolated dominant bacteria is identified according to the morphological feature and physiological and biochemical property of bacterium；From the bacterium of bacterium
Fall form and bio-chemical characteristics determined dominant bacteria be Lactobacillus acidophilus, it is bacillus coagulans, bacillus licheniformis, withered
After careless bacillus, bacillus amyloliquefaciens, this 5 kinds of bacterium are subjected to single strain fermentation, then Lactobacillus acidophilus, condensation bud
Spore bacillus, bacillus licheniformis, bacillus subtilis, bacillus amyloliquefaciens mix in proportion, and bacterium number quantity ratio is 1.8-2.2
: 0.8-1: 1.8-2.2: 1.8-2.2: 0.8-1, total bacteria count reaches 10,000,000,000/g.
Certain preferred embodiments according to the present invention, in step S4, the Lactobacillus acidophilus, bacillus coagulans, lichens
Bacillus, bacillus subtilis, bacillus amyloliquefaciens bacterium number quantity ratio be 2: 1: 2: 2: 1.
Embodiment 1
A kind of complex microorganism preparations adjusting aquatic livestock enteron aisle, the component including following bacterium number quantitative proportion: acidophilus
Bacillus acidi lactici, bacillus coagulans, bacillus licheniformis, bacillus subtilis, bacillus amyloliquefaciens, bacterium number quantity ratio are 2:
1: 2: 2: 1, the additive amount in breeding water body is 5ppm, and the additive amount in feed is 0.8%.
Above-mentioned complex microorganism preparations the preparation method is as follows:
S1, screened from grass carp body, dissect grass carp, take out entire enteron aisle, with 4 DEG C of sterile saline repeated flushing, put
Enter in homogenizer, is homogenized, takes 1.0g sample, be put into equipped with 9mL sterile water and be placed in the 50mL triangular flask of bead, set shaking table
Upper oscillation 20min disperses microbial cell, stand 30s 10-1Dilution；
S2,10 are drawn with 1mL aseptic straw-1Dilution 1mL is moved into the test tube equipped with 9mL sterile water, and suction is blown 3 times, is mixed
Combined bacteria liquid, 10-2Dilution；And so on, serial dilution is made 10-3、10-4、10-5、10-6、10-7、10-8It is a series of
Dilute bacterium solution；
S3, from 10-6、10-7、10-8It respectively takes 1mL in plate in dilution bacterium solution, pours into respectively and melt and be cooled to 45-50
DEG C nutrient broth agar culture medium and MRS culture medium, gently pivotal plate, is uniformly mixed bacterium solution with culture medium；After cooling
It is inverted, is observed after culture 24-48 hours in 28-30 DEG C；The bacterial strain separated from solid plate is if not single bacterium breeding
It forms, then further progress isolates and purifies；Well-grown representational colony inoculation inclined-plane is selected on plate, is done simultaneously
Plate coating checking, if discovery is impure, picking colony is further separated, until obtaining pure culture body；
S4, isolated dominant bacteria is identified according to the morphological feature and physiological and biochemical property of bacterium；From the bacterium of bacterium
Fall form and bio-chemical characteristics determined dominant bacteria be Lactobacillus acidophilus, it is bacillus coagulans, bacillus licheniformis, withered
After careless bacillus, bacillus amyloliquefaciens, this 5 kinds of bacterium are subjected to single strain fermentation, then Lactobacillus acidophilus, condensation bud
Spore bacillus, bacillus licheniformis, bacillus subtilis, bacillus amyloliquefaciens mix in proportion, and bacterium number quantity ratio is 2: 1: 2: 2
: 1, total bacteria count reaches 10,000,000,000/g.
Comparative example 1
Embodiment 1 is repeated, the difference is that: a kind of complex microorganism preparations adjusting aquatic livestock enteron aisle, including
The component of following bacterium number quantitative proportion: Lactobacillus acidophilus, bacillus coagulans, bacillus licheniformis, bacillus subtilis, bacterium
Number quantity ratio is 2: 1: 2: 2, and the additive amount in breeding water body is 5ppm.
Comparative example 2
Embodiment 1 is repeated, the difference is that: a kind of complex microorganism preparations adjusting aquatic livestock enteron aisle, including
The component of following bacterium number quantitative proportion: Lactobacillus acidophilus, bacillus coagulans, bacillus licheniformis, bacillus amyloliquefaciens,
Bacterium number quantity ratio is 2: 1: 2: 1, and the additive amount in feed is 0.8%.
Comparative example 3
Embodiment 1 is repeated, the difference is that: a kind of complex microorganism preparations adjusting aquatic livestock enteron aisle, including
The component of following bacterium number quantitative proportion: Lactobacillus acidophilus, bacillus coagulans, bacillus subtilis, bacillus amyloliquefaciens,
Bacterium number quantity ratio is 2: 1: 2: 1, and the additive amount in feed is 0.8%.
Comparative example 4
Embodiment 1 is repeated, the difference is that: a kind of complex microorganism preparations adjusting aquatic livestock enteron aisle, including
The component of following bacterium number quantitative proportion: Lactobacillus acidophilus, bacillus licheniformis, bacillus subtilis, bacillus amyloliquefaciens,
Bacterium number quantity ratio is 2: 2: 2: 1, and the additive amount in feed is 0.8%.
Comparative example 5
Embodiment 1 is repeated, the difference is that: a kind of complex microorganism preparations adjusting aquatic livestock enteron aisle, including
The component of following bacterium number quantitative proportion: Lactobacillus acidophilus, bacillus coagulans bacterium number quantity ratio be 2: 1, in breeding water body
In additive amount be 5ppm, additive amount in feed is 0.8%.
Comparative example 6
Embodiment 1 is repeated, the difference is that: a kind of complex microorganism preparations adjusting aquatic livestock enteron aisle, including
The component of following bacterium number quantitative proportion: Lactobacillus acidophilus, bacillus subtilis, bacillus amyloliquefaciens, bacterium number quantity ratio are 2
: 2: 1, the additive amount in breeding water body is 5ppm, and the additive amount in feed is 0.8%.
Comparative example 7
Embodiment 1 is repeated, the difference is that: a kind of complex microorganism preparations adjusting aquatic livestock enteron aisle, including
The component of following bacterium number quantitative proportion: Lactobacillus acidophilus, bacillus licheniformis, bacillus amyloliquefaciens, bacterium number quantity ratio are 2
: 2: 1, the additive amount in breeding water body is 5ppm, and the additive amount in feed is 0.8%.
Comparative example 8
Embodiment 1 is repeated, the difference is that: a kind of complex microorganism preparations adjusting aquatic livestock enteron aisle, including
The component of following bacterium number quantitative proportion: bacillus licheniformis, bacillus subtilis, bacillus amyloliquefaciens, bacterium number quantity ratio are 2
: 2: 1, the additive amount in feed is 0.8%.
Comparative example 9
Embodiment 1 is repeated, the difference is that: a kind of complex microorganism preparations adjusting aquatic livestock enteron aisle, including
The component of following bacterium number quantitative proportion: bacillus subtilis, bacillus amyloliquefaciens, bacterium number quantity ratio is 2: 1, in feed
Additive amount is 0.8%.
Laboratory report:
1, Zhanjiang shrimp culture pond
1) three groups: blank group, control group, of the present invention group are set.
Blank group does not launch any inorganic agent；
Control group 1 launches 1 microorganism formulation of comparative example, and control group 2 launches 5 microorganism formulation of comparative example, and control group 3 is launched
6 microorganism formulation of comparative example, control group 4 launch 7 microorganism formulation of comparative example；
Of the present invention group of dispensing 1 microorganism formulation of embodiment.
2) result: it is shown in Table 1, of the present invention group of NH3- N, nitrite, COD are significantly lower than blank group and control group.
1 each group NH of table3The indexs such as-N, nitrite, COD
| Blank group | Control group 1 | Control group 2 | Control group 3 | Control group 4 | Of the present invention group | |
| NH3-N(mg/L) | 1.29 | 0.62 | 0.99 | 0.81 | 0.83 | 0.03 |
| Nitrite (mg/L) | 2.98 | 1.56 | 2.17 | 1.73 | 1.82 | 0.04 |
| COD(mg/L) | 70.02 | 40.34 | 56.20 | 46.12 | 47.07 | 19.76 |
Note: NH3- N measurement uses Berthelot spectrophotometry；Nitrous nitrification uses N- (1- naphthalene)-ethylenediamine
Photometry；COD measurement is measured using COD Quick testing instrument.
2, Tianjin Hangu shrimp culture pond
1) three groups: blank group, control group, of the present invention group are set.
Blank group does not launch any inorganic agent；
Control group 1 launches 2 microorganism formulation of comparative example, and control group 2 launches 5 microorganism formulation of comparative example, and control group 3 is launched
6 microorganism formulation of comparative example, control group 4 launch 8 microorganism formulation of comparative example；
Of the present invention group of dispensing 1 microorganism formulation of embodiment.
2) result: extracting the serum of each group prawn, and the immune indexes such as measurement Phenoloxidase Activities, antalzyme activity are shown in Table
2, the enzyme activity of table 3, of the present invention group of prawn is substantially better than blank group and control group.
2 each group Phenoloxidase Activities of table (unit: U/mg)
| 10th day | 20th day | 30th day | |
| Blank group | 1.7320 | 1.7156 | 1.7432 |
| Control group 1 | 1.7934 | 1.8678 | 1.8956 |
| Control group 2 | 1.7509 | 1.7878 | 1.7960 |
| Control group 3 | 1.7801 | 1.8138 | 1.8265 |
| Control group 4 | 1.7788 | 1.7972 | 1.8052 |
| Of the present invention group | 2.5325 | 2.7210 | 2.9436 |
3 each group antalzyme activity of table (unit: U/ml)
Blank group: 30 days average growth rates 261.4%, survival rate 79.3% are fed.
Control group 1: 30 days average growth rates 285.9%, survival rate 85.6% are fed.
Control group 2: 30 days average growth rates 265.5%, survival rate 82.3% are fed.
Control group 3: 30 days average growth rates 274.0%, survival rate 84.2% are fed.
Control group 4: 30 days average growth rates 272.8%, survival rate 83.5% are fed.
Of the present invention group: feeding 30 days average growth rates 318.2%, survival rate 92.1%.
Of the present invention group of prawn growth conditions are substantially better than blank group and control group.
3, grass carp aquaculture pond
1) three groups: blank group, control group, of the present invention group are set.
Blank group does not launch any inorganic agent；
Control group 1 launches 3 microorganism formulation of comparative example, and control group 2 launches 5 microorganism formulation of comparative example, and control group 3 is launched
7 microorganism formulation of comparative example, control group 4 launch 8 microorganism formulation of comparative example；
Of the present invention group of dispensing embodiment 1.
2) result: each group grass carp venous blood collection takes upper serum measurement activity of catalase, superoxide dismutase living
The immune indexes such as power, are shown in Table 4, table 5, and the enzyme activity of of the present invention group of prawn is substantially better than blank group and control group.
4 each group activity of catalase of table (unit: U/mL)
| 10th day | 20th day | 30th day | |
| Blank group | 4.3701 | 4.3863 | 4.3892 |
| Control group 1 | 10.6896 | 12.5785 | 13.5685 |
| Control group 2 | 6.4573 | 7.3580 | 7.4241 |
| Control group 3 | 8.9642 | 10.1536 | 10.8789 |
| Control group 4 | 8.6784 | 9.6886 | 10.1232 |
| Of the present invention group | 15.5533 | 20.4542 | 22.7446 |
5 each group superoxide dismutase activity of table (unit: U/mL)
| 10th day | 20th day | 30th day | |
| Blank group | 74.4832 | 75.2845 | 75.1845 |
| Control group 1 | 90.8982 | 94.5392 | 98.2956 |
| Control group 2 | 76.7892 | 77.6483 | 79.7891 |
| Control group 3 | 81.7482 | 84.5278 | 88.2734 |
| Control group 4 | 79.7893 | 83.7893 | 86.8597 |
| Of the present invention group | 103.7284 | 110.7802 | 123.5483 |
4, grass carp aquaculture pond
1) three groups: blank group, control group, of the present invention group are set.
Blank group does not launch any inorganic agent；
Control group 1 launches 4 microorganism formulation of comparative example, and control group 2 launches 5 microorganism formulation of comparative example, and control group 3 is launched
7 microorganism formulation of comparative example, control group 4 launch 9 microorganism formulation of comparative example；
Of the present invention group of dispensing embodiment 1.
2) result: taking grass carp anterior intestine, is homogenized centrifuging and taking supernatant, the immune fingers such as measurement lipase activity, amylase activity
Mark, is shown in Table 6, table 7, the enzyme activity of of the present invention group of prawn is substantially better than blank group and control group.
6 each group lipase activity of table (unit: U/g)
| 10th day | 20th day | 30th day | |
| Blank group | 2.3742 | 2.3899 | 2.3826 |
| Control group 1 | 2.7586 | 3.1202 | 3.7593 |
| Control group 2 | 2.4648 | 2.6235 | 2.8727 |
| Control group 3 | 2.5325 | 2.8948 | 3.2844 |
| Control group 4 | 2.4185 | 2.5700 | 2.7627 |
| Of the present invention group | 3.0177 | 4.2535 | 4.7551 |
7 each group amylase activity of table (unit: U/mg)
In summary: using Lactobacillus acidophilus of the invention, bacillus coagulans, lichens gemma bar according to special ratios
Bacterium, bacillus subtilis, bacillus amyloliquefaciens strain combination compounded, than be used alone it is a certain, certain is used alone
It is a kind of or certain several combined effect is used alone to get well.
Obviously, the above embodiment of the present invention be only to clearly illustrate example of the present invention, and not be pair
The restriction of embodiments of the present invention.For those of ordinary skill in the art, may be used also on the basis of the above description
To make other variations or changes in different ways.Here all embodiments can not be exhaustive.It is all to belong to this hair
The obvious changes or variations that bright technical solution is extended out are still in the scope of protection of the present invention.
Claims (5)
1. a kind of complex microorganism preparations for adjusting aquatic livestock enteron aisle, it is characterised in that: the complex microorganism preparations are by thermophilic
Yogurt acidfast bacilli, bacillus coagulans, bacillus licheniformis, bacillus subtilis, bacillus amyloliquefaciens composition, bacterium number quantity
Than being 1.8-2.2: 0.8-1: 1.8-2.2: 1.8-2.2: 0.8-1, total bacteria count reaches 10,000,000,000/g.
2. the complex microorganism preparations for adjusting aquatic livestock enteron aisle according to claim 1, it is characterised in that: acidophilus cream
Acidfast bacilli, bacillus coagulans, bacillus licheniformis, bacillus subtilis, bacillus amyloliquefaciens bacterium number quantity ratio be 2: 1
∶2∶2∶1。
3. the complex microorganism preparations for adjusting aquatic livestock enteron aisle according to claim 1, it is characterised in that: described multiple
Closing additive amount of the microorganism formulation in breeding water body is 3-8ppm, and the additive amount in feed is 0.5-1%.
4. the preparation method of the complex microorganism preparations of aquatic livestock enteron aisle is adjusted as described in any in the claims 1-3,
It is characterized by comprising the following steps:
S1, screened from grass carp body, dissect grass carp, take out entire enteron aisle, with 3-5 DEG C of sterile saline repeated flushing, be put into
In homogenizer, homogenate takes 1.0g sample, is put into equipped with 9mL sterile water and is placed in the 50mL triangular flask of bead, set on shaking table
Oscillation 18-22min disperses microbial cell, stands 25-35 seconds up to 10-1Dilution；
S2,10 are drawn with 1mL aseptic straw-1Dilution 1mL is moved into the test tube equipped with 9mL sterile water, and suction blows 3 times, Mixed Microbes
Liquid, 10-2Dilution；And so on, serial dilution is made 10-3、10-4、10-5、10-6、10-7、10-8A series of dilution
Bacterium solution；
S3, from 10-6、10-7、10-8It respectively takes 1mL in plate in dilution bacterium solution, pours into respectively and melt and be cooled to 45-50 DEG C
Nutrient broth agar culture medium and MRS culture medium, gently pivotal plate, is uniformly mixed bacterium solution with culture medium；It is inverted after cooling,
It is observed after culture 24-48 hours in 28-30 DEG C；The bacterial strain separated from solid plate if not single bacterium breeds,
Then further progress isolates and purifies；Well-grown representational colony inoculation inclined-plane is selected on plate, while making smear
It checks, if discovery is impure, picking colony is further separated, until obtaining pure culture body；
S4, isolated dominant bacteria is identified according to the morphological feature and physiological and biochemical property of bacterium；From the bacterium colony shape of bacterium
State and bio-chemical characteristics have determined that dominant bacteria is Lactobacillus acidophilus, bacillus coagulans, bacillus licheniformis, withered grass bud
After spore bacillus, bacillus amyloliquefaciens, this 5 kinds of bacterium are subjected to single strain fermentation, then Lactobacillus acidophilus, condensation gemma bar
Bacterium, bacillus licheniformis, bacillus subtilis, bacillus amyloliquefaciens mix in proportion, and bacterium number quantity ratio is 1.8-2.2:
0.8-1: 1.8-2.2: 1.8-2.2: 0.8-1, total bacteria count reaches 10,000,000,000/g.
5. adjusting the preparation method of the complex microorganism preparations of aquatic livestock enteron aisle according to claim 4, it is characterised in that:
In step S4, the Lactobacillus acidophilus, bacillus coagulans, bacillus licheniformis, bacillus subtilis, solution starch gemma bar
The bacterium number quantity ratio of bacterium is 2: 1: 2: 2: 1.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111939180A (en) * | 2019-05-17 | 2020-11-17 | 上海绿奥生物科技有限公司 | Compound preparation for treating enteritis of turbot and preparation method thereof |
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| CN111642620A (en) * | 2020-07-08 | 2020-09-11 | 漯河医学高等专科学校 | Compound probiotic preparation and preparation method and application thereof |
| CN111909868A (en) * | 2020-07-24 | 2020-11-10 | 天津开发区坤禾生物技术有限公司 | Composite microbial inoculum for preventing and treating anorexia dead fish of grass carps as well as preparation method and application thereof |
| CN112501087A (en) * | 2020-12-23 | 2021-03-16 | 南京升鳌生物环保科技有限公司 | EM (effective microorganisms) regulator for improving intestinal flora of cultured aquatic products and preparation method and application thereof |
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