CN115261250A - Preparation method of synbiotics for shrimp and crab intestinal health - Google Patents
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Abstract
The invention discloses a preparation method of a synbiotics for shrimp and crab intestinal health, which comprises the following steps: firstly, screening bacillus coagulans and lactobacillus; completing the activation treatment of the strains; the strain is rated on the substrate; the strain carries out fermentation treatment on the substrate; GPX, SOD, NADP peroxidase, active polypeptide, active polysaccharide, organic acid, short chain fatty acid, and thallus components such as peptidoglycan, teichoic acid, lipoteichoic acid, etc. are generated during fermentation; compounding lactulose montmorillonite material; the product is evaluated by animal experiments, thereby promoting animal immunity and disease resistance; the synbiotics for shrimp and crab intestinal health are prepared to generate rich GPX, SOD, NADP peroxidase, active polypeptide, active polysaccharide, organic acid, short-chain fatty acid, and active substances such as peptidoglycan, teichoic acid, lipoteichoic acid and the like which are components of thalli.
Description
Technical Field
The invention relates to the technical field of aquaculture, in particular to a preparation method of a synbiotics for shrimp and crab intestinal tract health.
Background
Freshwater shrimp and crab culture is a key block of aquaculture industry in China, and comprises a plurality of varieties, wherein the main varieties are river crabs (eriocheir sinensis), macrobrachium rosenbergii, penaeus vannamei boone, freshwater shrimps and the like. Freshwater shrimps and crabs are one of the important components of famous and high-quality aquatic products in China, the cultivation of the freshwater shrimps and crabs starts in the seventies, the cultivation capacity is continuously increased along with the enlargement of the cultivation area, and the problems of overlarge reproduction density, reduced quality of germplasm resources and the like are caused by pursuing economic benefits and neglecting the importance of sustainable development, so that the water quality of the cultivation water body is deteriorated, the resistance of aquatic animals is reduced, opportunities are provided for the invasion of pathogenic bacteria, and particularly, the freshwater shrimps and crabs invade the intestinal tracts of the shrimps and crabs, and the phenomenon that intestinal diseases frequently occur is caused. The aquaculture sewage generated in the aquaculture process can further cause secondary pollution to the surrounding water body, and the abuse of antibiotics and pesticides can also cause chemical pollution, which can cause diseases by influencing the intestinal health of the shrimps and crabs. The economic loss caused by diseases reaches several billion yuan every year. Regarding the above phenomena, the aquaculture personnel usually take a lot of medicines to treat diseases as the final purpose, and do not take the factors of healthy cultivation, food safety, environmental friendliness and the like into consideration comprehensively. Therefore, the diseases are often incorrectly diagnosed, scientific medication is difficult, the medication for the opposite party is difficult, the medicine selection has no clear indication, a plurality of medicines are used simultaneously, a plurality of factors such as the curative effect of the medicines are not considered, the interference possibly caused by the factors on the medicines is not fully considered, and the like, and the concentration of medication, the times of medication, the interval time of medication and the like are not reasonable. The methods interfere with the rule of in vivo metabolism of the shrimps and the crabs, not only can the problem of diseases of the shrimp and the crabs not be solved fundamentally, but also damage the intestinal health of the shrimps and the crabs and influence the breeding benefit and even the food safety.
Aiming at the intestinal health problems of shrimps and crabs, probiotics, prebiotics and synbiotics are good solutions. Probiotics proliferate in the intestinal tract and develop microorganisms that have a positive health impact. The probiotic bacteria beneficial to intestinal tracts of shrimps and crabs comprise lactobacillus acidophilus, lactobacillus bulgaricus, bifidobacterium, enterococcus faecalis and the like. They can colonize the digestive tract of host and play corresponding physiological roles, such as increasing antibody titer in organism, enhancing macrophage activity, increasing the number of killer T cells, increasing interferon level and enhancing organism immunity. Prebiotics are oligosaccharides or polysaccharides that can be used as substrates by normal intestinal flora, selectively stimulate the growth or increase the activity of one or more bacteria in the colon, and are beneficial to the health of the host, such as fructooligosaccharides, galactooligosaccharides, isomaltooligosaccharides, soy oligosaccharides, and the like. The synbiotics refer to a mixed product of probiotics and prebiotics, or other substances beneficial to intestinal health are added. It can exert physiological bacterial activity of the probiotics, and selectively increase the number of the probiotics, so that the effect of the probiotics is more remarkable and lasting. However, few synbiotics research is conducted at home and abroad, and the functions of the synbiotics are still to be further researched.
Disclosure of Invention
The invention aims to provide a preparation method of a synbiotics for shrimp and crab intestinal health aiming at the defects of the prior art, and aims to solve the problems in the background art.
In order to achieve the purpose, the invention provides the following technical scheme: the method comprises the following steps:
the method comprises the following steps: firstly, screening bacillus coagulans and lactobacillus;
step two: completing the activation treatment of the strain;
step three: the strain is rated on a substrate;
step four: the strain performs fermentation treatment on the substrate;
step five: GPX, SOD, NADP peroxidase, active polypeptide, active polysaccharide, organic acid, short chain fatty acid, and thallus components such as peptidoglycan, teichoic acid, lipoteichoic acid, etc. are generated during fermentation;
step six: compounding lactulose montmorillonite material;
step seven: the product is evaluated by animal experiment, thereby promoting animal immunity and disease resistance.
As a preferred technical scheme of the invention, the bacillus coagulans is separated from nature and selected by markets, and the strains meet the following conditions: the survival rate of the bacillus coagulans strain is more than 98 percent after being treated at the high temperature of 100 ℃ for 5 min; the survival rate of the bacillus coagulans strain is more than 96 percent after being treated by medium-strong acid liquid for 2 hours; the first generation of propagation is completed within 30min and the propagation is completed by more than 300 times within 4 hours when the bacillus coagulans strain is propagated; the bacillus coagulans strain is stored for 2 years under natural conditions, and the number of live bacteria is not obviously changed.
As a preferred technical scheme of the invention, the lactobacillus is separated from nature and selected by the market, and the strains meet the following regulation: the lactobacillus strain grows under the condition of pH3.0, the lactobacillus strain has obvious bacteriostatic action on Escherichia coli and staphylococcus aureus, a large amount of organic acid is rapidly produced after the lactobacillus strain is fermented for 2 hours, and the pH tends to be stable after the lactobacillus strain is fermented for 12 hours and is below 3.7.
As a preferred technical scheme of the invention, the Bacillus coagulans fermentation substrate and conditions are as follows: the wheat bran is used as a substrate, the content of the wheat bran crude protein is more than 13%, and 0.8-1.5% of carbon source (glucose, lactose, sucrose and glycerol), nitrogen source (ammonium salt, ammonium chloride, sodium nitrate and peptone) and salt substance (KH 2P04-1.0%, naH2P04-0.85%, K2HP04-1.5%, na2HP04-1.2% and MnS 04-0.1%) are added into the substrate; humidity of 55% to 70%; the pH value is 6.5-7.0.
As a preferred technical scheme of the invention, the lactobacillus fermentation substrate and conditions are as follows: the soybean powder is used as a substrate, the content of the crude protein of the soybean powder is more than 30 percent, and a2 percent nutrient medium (the formula is that peptone is 12.0g, beef powder is 5.0g, yeast extract is 5.0g, glucose is 25.0g, tween-80.1ml, dipotassium hydrogen phosphate is 2.0g, sodium acetate is 4.5g, triammonium citrate is 2.0g, magnesium sulfate heptahydrate is 0.2g and manganese sulfate tetrahydrate is 0.05 g) is added into the substrate.
As a preferable technical scheme of the invention, the montmorillonite is natural sodium montmorillonite, the ion exchange amount of the montmorillonite is within the range of 100meq/100g to 200meq/100g, and the purity of lactulose is more than 99.0%.
Compared with the prior art, the invention provides a preparation method of a synbiotics for shrimp and crab intestinal health, which has the following beneficial effects:
the invention relates to a preparation method of synbiotics for shrimp and crab intestinal health, which is characterized in that bacillus coagulans and lactobacillus are screened, activated, planted, propagated and fermented on a substrate to obtain active substances such as GPX, SOD, NADP peroxidase, active polypeptide, active polysaccharide, organic acid, short-chain fatty acid, constituent components of bacteria, namely peptidoglycan, teichoic acid, lipoteichoic acid and the like, and the active substances are compounded by matching oligosaccharide montmorillonite. The concrete method is that the adsorption performance of the montmorillonite is utilized to adsorb and gather microorganisms; the prebiotic function of lactulose is utilized to provide nutrients for beneficial microorganisms in intestinal tracts, so that harmful microorganisms lose competitive advantages; the beneficial bacteria book in intestinal tracts is increased by utilizing bacillus coagulans and lactobacillus; the fermentation products of the bacillus coagulans and the lactobacillus provide nutrients for intestinal cells, the structure of the intestinal tract is optimized, and the function of the intestinal tract is improved.
The preparation method of the synbiotics for shrimp and crab intestinal health has the following characteristics:
1. systematically building an intestinal optimization scheme for the shrimps and crabs instead of simple sterilization and pathogenicity;
2. the product obtained by the preparation technology is a green intestinal health care agent, has no toxic or side effect, and has the function of replacing antibiotics;
3. the obtained material has good dispersibility, and is suitable for feed production and other uses.
Drawings
FIG. 1 is a schematic diagram of 120mg/Kg test of the present invention;
FIG. 2 is a schematic diagram of the 250mg/Kg test of the present invention;
FIG. 3 is a schematic diagram of the 500mg/Kg test of the present invention;
FIG. 4 is a schematic diagram of a 1000mg/Kg test of the present invention;
FIG. 5 is a schematic illustration of an assay according to the present invention;
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
The first embodiment is as follows: the product is added into penaeus vannamei feed according to different dosages, the comprehensive effect of 250g/T dosage is found to be optimal after the product is used, the intestinal structure and function can be regulated and controlled by optimizing intestinal flora, the intestinal digestive enzyme activity is improved, the protease activity is improved by 26.09%, the lipase activity is improved by 30.95%, the amylase activity is improved by 20.51%, and the intestinal digestive absorption capacity is finally improved.
The test evaluation was as follows:
in order to achieve the purpose, the invention provides the following technical scheme: the method comprises the following steps:
the method comprises the following steps: firstly, screening bacillus coagulans and lactobacillus;
step two: completing the activation treatment of the strains;
step three: the strain is rated on a substrate;
step four: the strain carries out fermentation treatment on the substrate;
step five: GPX, SOD, NADP peroxidase, active polypeptide, active polysaccharide, organic acid, short chain fatty acid, and thallus components such as peptidoglycan, teichoic acid, lipoteichoic acid, etc. are generated during fermentation;
step six: compounding lactulose montmorillonite material;
step seven: the product is evaluated by animal experiment, thereby promoting animal immunity and disease resistance.
Specifically, the method comprises the following steps: the bacillus coagulans is separated from nature and selected by markets, and the strains meet the following conditions: the survival rate of the bacillus coagulans strain is more than 98 percent after being treated at the high temperature of 100 ℃ for 5 min; the survival rate of the bacillus coagulans strain is more than 96 percent after being treated by medium-strong acid liquid for 2 hours; when the bacillus coagulans strain is propagated, one generation of propagation is completed within 30min and the propagation is completed by more than 300 times within 4 hours; the bacillus coagulans strain is stored for 2 years under natural conditions, and the number of live bacteria has no obvious change; the lactobacillus is separated by nature and selected by the market, and the strains meet the following regulation: the lactobacillus strain grows under the condition of pH3.0, has obvious bacteriostatic action on Escherichia coli and staphylococcus aureus, quickly generates a large amount of organic acid after fermenting for 2 hours, and has pH which is stable and is below 3.7 after fermenting for 12 hours; the bacillus coagulans fermentation substrate and conditions are as follows: wheat bran is used as a substrate, the content of wheat bran crude protein is more than 13%, 0.8-1.5% of carbon source (glucose, lactose, sucrose and glycerol), nitrogen source (ammonium salt, ammonium chloride, sodium nitrate and peptone) and salt substance (KH 2P04-1.0%, naH2P04-0.85%, K2HP04-1.5%, na2HP04-1.2% and MnS 04-0.1%) are added into the substrate; humidity of 55% to 70%; the PH value is 6.5-7.0; the lactobacillus fermentation substrate and conditions are as follows: soybean powder is used as a substrate, the content of crude protein of the soybean powder is more than 30 percent, and 2 percent of nutrient medium (the formula is that peptone is 12.0g, beef powder is 5.0g, yeast extract is 5.0g, glucose is 25.0g, tween-80.1ml, dipotassium hydrogen phosphate is 2.0g, sodium acetate is 4.5g, triammonium citrate is 2.0g, magnesium sulfate heptahydrate is 0.2g and manganese sulfate tetrahydrate is 0.05 g) is added into the substrate; the montmorillonite is natural sodium-based montmorillonite, the ion exchange amount of the montmorillonite is within the range of 100meq/100g to 200meq/100g, and the purity of lactulose is more than 99.0%.
Example two: aeromonas hydrophila is one of the most common bacterial diseases of aquatic animals, and a large number of deaths occur after aquatic animals suffer from the bacterial attack. The death rate of the macrobrachium rosenbergii in 24 hours is obviously lower than that of the macrobrachium rosenbergii without using the macrobrachium rosenbergii after the aeromonas hydrophila challenge experiment is carried out on the macrobrachium rosenbergii using the product. After 5 days of toxin attacking, the cumulative mortality rate of the macrobrachium rosenbergii using the product at 250g/T is reduced by about 30 percent compared with that of a control group.
The test evaluation is shown in FIGS. 1 to 5
In order to achieve the purpose, the invention provides the following technical scheme: the method comprises the following steps:
the method comprises the following steps: firstly, screening bacillus coagulans and lactobacillus;
step two: completing the activation treatment of the strain;
step three: the strain is rated on a substrate;
step four: the strain performs fermentation treatment on the substrate;
step five: GPX, SOD, NADP peroxidase, active polypeptide, active polysaccharide, organic acid, short chain fatty acid, and thallus components such as peptidoglycan, teichoic acid, lipoteichoic acid, etc. are generated during fermentation;
step six: compounding lactulose montmorillonite material;
step seven: the product is evaluated by animal test, thereby promoting animal immunity and disease resistance.
Specifically, the method comprises the following steps: the bacillus coagulans is separated by nature and selected by the market, and the strain meets the following conditions: the survival rate of the bacillus coagulans strain is more than 98 percent after being treated at the high temperature of 100 ℃ for 5 min; the survival rate of the bacillus coagulans strain is more than 96 percent after being treated by medium-strong acid liquid for 2 hours; when the bacillus coagulans strain is propagated, first-generation propagation is completed within 30min and the propagation is completed by more than 300 times within 4 hours; the bacillus coagulans strain is stored for 2 years under natural conditions, and the number of live bacteria has no obvious change; the lactobacillus is separated by nature and selected by the market, and the strains meet the following regulation: the lactobacillus strain grows under the condition of pH3.0, the lactobacillus strain has obvious bacteriostatic action on Escherichia coli and staphylococcus aureus, a large amount of organic acid is rapidly generated after the lactobacillus strain is fermented for 2 hours, and the pH tends to be stable after the lactobacillus strain is fermented for 12 hours and reaches below 3.7; the bacillus coagulans fermentation substrate and conditions are as follows: wheat bran is used as a substrate, the content of wheat bran crude protein is more than 13%, 0.8-1.5% of carbon source (glucose, lactose, sucrose and glycerol), nitrogen source (ammonium salt, ammonium chloride, sodium nitrate and peptone) and salt substance (KH 2P04-1.0%, naH2P04-0.85%, K2HP04-1.5%, na2HP04-1.2% and MnS 04-0.1%) are added into the substrate; humidity of 55% to 70%; the PH value is 6.5-7.0; the lactobacillus fermentation substrate and conditions are as follows: soybean powder is used as a substrate, the content of crude protein of the soybean powder is more than 30 percent, and 2 percent of nutrient medium (the formula is that peptone is 12.0g, beef powder is 5.0g, yeast extract is 5.0g, glucose is 25.0g, tween-80.1ml, dipotassium hydrogen phosphate is 2.0g, sodium acetate is 4.5g, triammonium citrate is 2.0g, magnesium sulfate heptahydrate is 0.2g and manganese sulfate tetrahydrate is 0.05 g) is added into the substrate; the montmorillonite is natural sodium-based montmorillonite, the ion exchange amount of the montmorillonite is within the range of 100meq/100g to 200meq/100g, and the purity of lactulose is more than 99.0%.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described above, or equivalents may be substituted for elements thereof. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (6)
1. A preparation method of synbiotics for shrimp and crab intestinal health is characterized by comprising the following steps: the method comprises the following steps:
the method comprises the following steps: firstly, screening bacillus coagulans and lactobacillus;
step two: completing the activation treatment of the strain;
step three: the strain is rated on a substrate;
step four: the strain performs fermentation treatment on the substrate;
step five: GPX, SOD, NADP peroxidase, active polypeptide, active polysaccharide, organic acid, short chain fatty acid, and thallus components such as peptidoglycan, teichoic acid, lipoteichoic acid, etc. are generated during fermentation;
step six: compounding lactulose montmorillonite material;
step seven: the product is evaluated by animal test, thereby promoting animal immunity and disease resistance.
2. The method for preparing a synbiotic for shrimp and crab intestinal health according to claim 1, wherein the method comprises the following steps: in the first step: the bacillus coagulans is separated by nature and selected by the market, and the strain meets the following conditions: the survival rate of the bacillus coagulans strain is more than 98 percent after being treated at the high temperature of 100 ℃ for 5 min; the survival rate of the bacillus coagulans strain is more than 96 percent after being treated by medium-strong acid liquid for 2 hours; the first generation of propagation is completed within 30min and the propagation is completed by more than 300 times within 4 hours when the bacillus coagulans strain is propagated; the bacillus coagulans strain is stored for 2 years under natural conditions, and the number of live bacteria is not obviously changed.
3. The method for preparing a synbiotics for shrimp and crab intestinal health according to claim 1, which is characterized by comprising the following steps: in the second step: the lactobacillus is separated by nature and selected by the market, and the strains meet the following regulation: the lactobacillus strain grows under the condition of pH3.0, the lactobacillus strain has obvious bacteriostatic action on Escherichia coli and staphylococcus aureus, a large amount of organic acid is rapidly generated after the lactobacillus strain is fermented for 2 hours, and the pH tends to be stable after the lactobacillus strain is fermented for 12 hours and is below 3.7.
4. The method for preparing a synbiotics for shrimp and crab intestinal health according to claim 1, which is characterized by comprising the following steps: in the third step, the bacillus coagulans fermentation substrate and conditions are as follows: the wheat bran is used as a substrate, the content of the wheat bran crude protein is more than 13%, and 0.8-1.5% of carbon sources (glucose, lactose, sucrose and glycerol) and nitrogen sources (ammonium salt, ammonium chloride, sodium nitrate and peptone) and salt substances (KH 2P04-1.0%, naH2P04-0.85%, K2HP04-1.5%, na2HP04-1.2% and MnS 04-0.1%) are added into the substrate; humidity of 55% to 70%; the pH value is 6.5-7.0.
5. The method for preparing a synbiotics for shrimp and crab intestinal health according to claim 1, which is characterized by comprising the following steps: in the third step, the lactobacillus fermentation substrate and conditions are as follows: the soybean powder is used as a substrate, the content of the crude protein of the soybean powder is more than 30 percent, and a2 percent nutrient medium (the formula is that peptone is 12.0g, beef powder is 5.0g, yeast extract is 5.0g, glucose is 25.0g, tween-80.1ml, dipotassium hydrogen phosphate is 2.0g, sodium acetate is 4.5g, triammonium citrate is 2.0g, magnesium sulfate heptahydrate is 0.2g and manganese sulfate tetrahydrate is 0.05 g) is added into the substrate.
6. The method for preparing a synbiotics for shrimp and crab intestinal health according to claim 1, which is characterized by comprising the following steps: the montmorillonite is natural sodium-based montmorillonite, the ion exchange amount of the montmorillonite is within the range of 100meq/100g to 200meq/100g, and the purity of lactulose is more than 99.0%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210468682.3A CN115261250A (en) | 2022-04-29 | 2022-04-29 | Preparation method of synbiotics for shrimp and crab intestinal health |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1663573A (en) * | 2004-03-04 | 2005-09-07 | 青岛东海药业有限公司 | A stable and safe microecological formulation, its preparation and usage |
| CN104017751A (en) * | 2014-04-17 | 2014-09-03 | 青岛九和宜生生物科技有限公司 | Probiotics prepared from bacillus coagulans and lactobacillus plantarum as well as preparation method thereof |
| CN110974845A (en) * | 2019-12-25 | 2020-04-10 | 苏州华健瑞达医药技术有限公司 | Pharmaceutical composition for improving gastrointestinal health, preparation method thereof and preparation method of pharmaceutical preparation |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1663573A (en) * | 2004-03-04 | 2005-09-07 | 青岛东海药业有限公司 | A stable and safe microecological formulation, its preparation and usage |
| CN104017751A (en) * | 2014-04-17 | 2014-09-03 | 青岛九和宜生生物科技有限公司 | Probiotics prepared from bacillus coagulans and lactobacillus plantarum as well as preparation method thereof |
| CN110974845A (en) * | 2019-12-25 | 2020-04-10 | 苏州华健瑞达医药技术有限公司 | Pharmaceutical composition for improving gastrointestinal health, preparation method thereof and preparation method of pharmaceutical preparation |
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