CN111759950A - Composite probiotic fermentation product for preventing and treating aquatic vibriosis - Google Patents
Composite probiotic fermentation product for preventing and treating aquatic vibriosis Download PDFInfo
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Abstract
The invention discloses a composite probiotic fermentation product for preventing and treating aquatic vibriosis, which is used in the field of aquatic vibriosis prevention and treatment, wherein the fermentation formula of the fermentation product comprises the following components in percentage by mass: 6-12% of scutellaria baicalensis, 3-10% of eucalyptus leaves, 5-7% of dendrobium officinale, 2-4% of bacillus subtilis liquid, 2-4% of candida utilis liquid, 15-20% of corn flour, 15-20% of soybean meal powder, 5-8% of wheat bran, 5-8% of brown sugar and 15-20% of water; the fermentation conditions of the fermentation product are as follows: fermenting at 25-28 deg.C and 50-100r/min for 8-12 days under initial pH of 5-6. The fermentation product obtained by the invention has strong killing and inhibiting effects on vibrio alginolyticus, vibrio anguillarum and vibrio parahaemolyticus of vibrio.
Description
Technical Field
The invention relates to the field of prevention and treatment of aquatic vibriosis, in particular to a composite probiotic fermentation product for preventing and treating aquatic vibriosis.
Background
The vibriosis is a common disease of seawater aquatic products, is easily damaged by the vibriosis during the culture of fish and shrimp seedlings, has the pathogeny mainly of vibrio bacteria, and has the main symptoms: diseased fish and shrimp are mainly characterized by body surface ulcer, and are rotten due to head osculum, body surface bleeding and visceral organ lesion is obvious sometimes. The disease occurs every month, the period of 7-8 months is a high incidence period, and the death rate can reach more than 80 percent when the death rate is high. The current commonly used treatment means is to mix antibacterial drugs such as norfloxacin, ciprofloxacin, chloramphenicol, gentamicin and the like into bait, the dosage of the antibacterial drugs is 1-3 percent of the bait dosage, and the antibacterial drugs are continuously fed for 5-10 days. However, the use of antibiotics in large quantities not only can lead to the abuse of antibiotics and influence the ecological environment, but also can finally influence the health of human beings, thus being not beneficial to the healthy and sustainable development of the aquaculture industry of large water products.
The microbial fermentation Chinese herbal medicine integrates the advantages of Chinese herbal medicines, enzyme preparations, probiotics and the like, and has the advantages of being natural, non-toxic, free of side effects, free of residues, safe and clean. The effective components of the Chinese herbal medicine are very complex, mainly comprise alkaloid, saponins, volatile oil, flavonoid, polysaccharide, organic acid, mineral elements and vitamins, and the active components are beneficial to enhancing the immune function, the anti-stress function, the antibacterial function and the like of the organism.
Aiming at the problems of antibiotic abuse and ecological influence caused by the use of antibiotics at present, the inventor develops a product of a compound probiotic fermented traditional Chinese medicine for preventing and treating vibriosis in aquatic products.
Disclosure of Invention
Aiming at the problems and defects in the prior art, the invention provides a composite probiotic fermentation product for preventing and treating vibriosis in aquatic products.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a composite probiotic fermentation product for preventing and treating vibriosis in aquatic products, wherein the fermentation formula of the fermentation product comprises the following components in percentage by mass: 6-12% of scutellaria baicalensis, 3-10% of eucalyptus leaves, 5-7% of dendrobium officinale, 2-4% of bacillus subtilis liquid, 2-4% of candida utilis liquid, 15-20% of corn flour, 15-20% of soybean meal powder, 5-8% of wheat bran, 5-8% of brown sugar and 15-20% of water; the fermentation conditions of the fermentation product are as follows: fermenting at 25-28 deg.C and 50-100r/min for 8-12 days under initial pH of 5-6.
Preferably, the fermentation formula of the fermentation product comprises the following components in percentage by mass: 10% of scutellaria baicalensis, 10% of eucalyptus leaves, 7% of dendrobium officinale, 3% of bacillus subtilis liquid, 3% of candida utilis liquid, 17% of corn flour, 20% of soybean meal, 5% of wheat bran, 5% of brown sugar and 20% of water; the fermentation conditions of the fermentation product are as follows: fermenting for 12 days under the conditions of initial pH value of 6, temperature of 25 ℃ and rotation speed of 100 r/min.
Preferably, the fermentation formula of the fermentation product comprises the following components in percentage by mass: 12% of scutellaria baicalensis, 8% of eucalyptus leaves, 6% of dendrobium officinale, 4% of bacillus subtilis liquid, 4% of candida utilis liquid, 16% of corn flour, 20% of soybean meal, 5% of wheat bran, 5% of brown sugar and 20% of water; the fermentation conditions of the fermentation product are as follows: fermenting at 28 deg.C and 100r/min for 10 days under initial pH of 5.5.
Preferably, the preparation method of the bacillus subtilis liquid comprises the following steps:
uniformly mixing 10g of peptone, 3g of beef extract, 10ml of glucose, 5g of sodium chloride, 10ml of 3.08% magnesium sulfate and 1L of water, and sterilizing for 20 minutes at the temperature of 121 ℃ under the pressure of 0.15MPa by using high-pressure steam to obtain a liquid culture medium for later use;
② inoculating the screened and purified Bacillus subtilis strain to liquid culture medium according to 9% of inoculation amount, performing propagation, performing static culture at 37 deg.C for 24 hr, and adjusting effective viable count to 1x109cfu/ml to obtain the bacillus subtilis liquid.
Preferably, the preparation method of the candida utilis bacterial liquid comprises the following steps:
uniformly mixing 10g of yeast extract powder, 20g of peptone and 20g of glucose, then using distilled water to fix the volume to 1L, and sterilizing for 20 minutes by high-pressure steam at the temperature of 121 ℃ under the pressure of 0.15MPa to obtain a liquid culture medium for later use;
② inoculating the screened and purified Candida utilis strain to liquid culture medium according to 9% of inoculation amount for propagation, performing static culture at 37 ℃ for 24 hours, and adjusting the effective viable count to 1x109cfu/ml to obtain candida utilis bacterial liquid.
By adopting the technical scheme, the product of the scutellaria baicalensis, the eucalyptus leaves and the dendrobium officinale through composite fermentation of the candida utilis and the bacillus subtilis has an inhibition zone of 16.5-17.1mm on various vibrio bacteria, and can effectively prevent vibriosis in aquatic products.
Detailed Description
Here, the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
In the description of the present invention, it should be noted that the terms "upper", "lower", "inner", "outer", "front", "rear", "both ends", "one end", "the other end", and the like indicate orientations or positional relationships only for convenience in describing the present invention and simplifying the description, but do not indicate or imply that the referred device or element must have a specific orientation, be constructed in a specific orientation, and be operated, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the description of the present invention, it should be noted that, unless otherwise explicitly stated or limited, the terms "mounted," "disposed," "connected," and the like are to be construed broadly, and for example, "connected" may be fixedly, detachably, or integrally connected; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meaning of the above terms in the present invention can be understood in specific cases to those skilled in the art.
In the first embodiment, a fermentation formula of a composite probiotic fermentation product for preventing and treating vibriosis in aquatic products comprises the following components in percentage by mass: 10% of scutellaria baicalensis, 10% of eucalyptus leaves, 7% of dendrobium officinale, 3% of bacillus subtilis liquid, 3% of candida utilis liquid, 17% of corn flour, 20% of soybean meal, 5% of wheat bran, 5% of brown sugar and 20% of water; the fermentation conditions of the fermentation product are as follows: fermenting for 12 days under the conditions of initial pH value of 6, temperature of 25 ℃ and rotation speed of 100 r/min.
The preparation method of the bacillus subtilis liquid comprises the following steps:
uniformly mixing 10g of peptone, 3g of beef extract, 10ml of glucose, 5g of sodium chloride, 10ml of 3.08% magnesium sulfate and 1L of water, and sterilizing for 20 minutes at the temperature of 121 ℃ under the pressure of 0.15MPa by using high-pressure steam to obtain a liquid culture medium for later use;
② inoculating the screened and purified Bacillus subtilis strain to liquid culture medium according to 9% of inoculation amount, performing propagation, performing static culture at 37 deg.C for 24 hr, and adjusting effective viable count to 1x109cfu/ml to obtain the bacillus subtilis liquid.
The preparation method of the candida utilis bacterial liquid comprises the following steps:
uniformly mixing 10g of yeast extract powder, 20g of peptone and 20g of glucose, then using distilled water to fix the volume to 1L, and sterilizing for 20 minutes by high-pressure steam at the temperature of 121 ℃ under the pressure of 0.15MPa to obtain a liquid culture medium for later use;
② inoculating the screened and purified Candida utilis strain to liquid culture medium according to 9% of inoculation amount for propagation, performing static culture at 37 ℃ for 24 hours, and adjusting the effective viable count to 1x109cfu/ml to obtain candida utilis bacterial liquid.
In the second embodiment, a composite probiotic fermentation product for preventing and treating vibriosis in aquatic products comprises the following components in percentage by mass: 12% of scutellaria baicalensis, 8% of eucalyptus leaves, 6% of dendrobium officinale, 4% of bacillus subtilis liquid, 4% of candida utilis liquid, 16% of corn flour, 20% of soybean meal, 5% of wheat bran, 5% of brown sugar and 20% of water; the fermentation conditions of the fermentation product are as follows: fermenting at 28 deg.C and 100r/min for 10 days at initial pH of 5.5.
The preparation method of the bacillus subtilis liquid comprises the following steps:
uniformly mixing 10g of peptone, 3g of beef extract, 10ml of glucose, 5g of sodium chloride, 10ml of 3.08% magnesium sulfate and 1L of water, and sterilizing for 20 minutes at the temperature of 121 ℃ under the pressure of 0.15MPa by using high-pressure steam to obtain a liquid culture medium for later use;
② inoculating the screened and purified Bacillus subtilis strain to liquid culture medium according to 9% of inoculation amount, performing propagation, performing static culture at 37 deg.C for 24 hr, and adjusting effective viable count to 1x109cfu/ml to obtain the bacillus subtilis liquid.
The preparation method of the candida utilis bacterial liquid comprises the following steps:
uniformly mixing 10g of yeast extract powder, 20g of peptone and 20g of glucose, then using distilled water to fix the volume to 1L, and sterilizing for 20 minutes by high-pressure steam at the temperature of 121 ℃ under the pressure of 0.15MPa to obtain a liquid culture medium for later use;
② inoculating the screened and purified Candida utilis strain to liquid culture medium according to 9% of inoculation amount for propagation, performing static culture at 37 ℃ for 24 hours, and adjusting the effective viable count to 1x109cfu/ml to obtain candida utilis bacterial liquid.
In the third embodiment, the fermentation formula of the composite probiotic fermentation product for preventing and treating the vibriosis in the aquatic products comprises the following components in percentage by mass: 12% of scutellaria baicalensis, 10% of eucalyptus leaves, 7% of dendrobium officinale, 4% of bacillus subtilis liquid, 4% of candida utilis liquid, 20% of corn flour, 17% of soybean meal, 5% of wheat bran, 5% of brown sugar and 16% of water; the fermentation conditions of the fermentation product are as follows: fermenting for 12 days under the conditions of initial pH value of 5, temperature of 28 ℃ and rotation speed of 100 r/min.
The preparation method of the bacillus subtilis liquid comprises the following steps:
uniformly mixing 10g of peptone, 3g of beef extract, 10ml of glucose, 5g of sodium chloride, 10ml of 3.08% magnesium sulfate and 1L of water, and sterilizing for 20 minutes at the temperature of 121 ℃ under the pressure of 0.15MPa by using high-pressure steam to obtain a liquid culture medium for later use;
② inoculating the screened and purified Bacillus subtilis strain to liquid culture medium according to 9% of inoculation amount, performing propagation, performing static culture at 37 deg.C for 24 hr, and adjusting effective viable count to 1x109cfu/ml to obtain the bacillus subtilis liquid.
The preparation method of the candida utilis bacterial liquid comprises the following steps:
uniformly mixing 10g of yeast extract powder, 20g of peptone and 20g of glucose, then using distilled water to fix the volume to 1L, and sterilizing for 20 minutes by high-pressure steam at the temperature of 121 ℃ under the pressure of 0.15MPa to obtain a liquid culture medium for later use;
② inoculating the screened and purified Candida utilis strain to liquid culture medium according to 9% of inoculation amount for propagation, performing static culture at 37 ℃ for 24 hours, and adjusting the effective viable count to 1x109cfu/ml to obtain candida utilis bacterial liquid.
In the fourth embodiment, the fermentation formula of the composite probiotic fermentation product for preventing and treating the vibrio disease of the aquatic products comprises the following components in percentage by mass: 11% of scutellaria baicalensis, 9% of eucalyptus leaves, 7% of dendrobium officinale, 3% of bacillus subtilis liquid, 3% of candida utilis liquid, 17% of corn flour, 20% of soybean meal, 5% of wheat bran, 5% of brown sugar and 20% of water; the fermentation conditions of the fermentation product are as follows: fermenting at the initial pH of 6, temperature of 26 deg.C and rotation speed of 100r/min for 11 days.
The preparation method of the bacillus subtilis liquid comprises the following steps:
uniformly mixing 10g of peptone, 3g of beef extract, 10ml of glucose, 5g of sodium chloride, 10ml of 3.08% magnesium sulfate and 1L of water, and sterilizing for 20 minutes at the temperature of 121 ℃ under the pressure of 0.15MPa by using high-pressure steam to obtain a liquid culture medium for later use;
② inoculating the screened and purified Bacillus subtilis strain to liquid culture medium according to 9% of inoculation amount, performing propagation, performing static culture at 37 deg.C for 24 hr, and adjusting effective viable count to 1x109cfu/ml to obtain the bacillus subtilis liquid.
The preparation method of the candida utilis bacterial liquid comprises the following steps:
uniformly mixing 10g of yeast extract powder, 20g of peptone and 20g of glucose, then using distilled water to fix the volume to 1L, and sterilizing for 20 minutes by high-pressure steam at the temperature of 121 ℃ under the pressure of 0.15MPa to obtain a liquid culture medium for later use;
② inoculating the screened and purified Candida utilis strain to liquid culture medium according to 9% of inoculation amount for propagation, performing static culture at 37 ℃ for 24 hours, and adjusting the effective viable count to 1x109cfu/ml to obtain candida utilis bacterial liquid.
Example five, a fermentation formulation of a complex probiotic fermentation product for controlling vibrio disease in aquatic products comprises the following components in percentage by mass: 10% of scutellaria baicalensis, 9% of eucalyptus leaves, 6% of dendrobium officinale, 4% of bacillus subtilis liquid, 4% of candida utilis liquid, 18% of corn flour, 18% of soybean meal, 6% of wheat bran, 6% of brown sugar and 19% of water; the fermentation conditions of the fermentation product are as follows: fermenting at 25-28 deg.C and 50-100r/min for 8-12 days under initial pH of 5-6.
The preparation method of the bacillus subtilis liquid comprises the following steps:
uniformly mixing 10g of peptone, 3g of beef extract, 10ml of glucose, 5g of sodium chloride, 10ml of 3.08% magnesium sulfate and 1L of water, and sterilizing for 20 minutes at the temperature of 121 ℃ under the pressure of 0.15MPa by using high-pressure steam to obtain a liquid culture medium for later use;
② inoculating the screened and purified Bacillus subtilis strain to liquid culture medium according to 9% of inoculation amount, performing propagation, performing static culture at 37 deg.C for 24 hr, and adjusting effective viable count to 1x109cfu/ml to obtain the bacillus subtilis liquid.
The preparation method of the candida utilis bacterial liquid comprises the following steps:
uniformly mixing 10g of yeast extract powder, 20g of peptone and 20g of glucose, then using distilled water to fix the volume to 1L, and sterilizing for 20 minutes by high-pressure steam at the temperature of 121 ℃ under the pressure of 0.15MPa to obtain a liquid culture medium for later use;
② inoculating the screened and purified Candida utilis strain to liquid culture medium according to 9% of inoculation amount for propagation, performing static culture at 37 ℃ for 24 hours, and adjusting the effective viable count to 1x109cfu/ml to obtain candida utilis bacterial liquid.
Example six, the fermentation products of examples one to five were used to perform zone tests on Vibrio alginolyticus, Vibrio anguillarum, and Vibrio parahaemolyticus, respectively, and the results of the zone tests are shown in the following table:
| vibrio alginolyticus | Vibrio anguillarum | Vibrio parahaemolyticus | |
| Example A radius of inhibition | 16.61mm | 16.50mm | 17.22mm |
| EXAMPLE two radius of inhibition | 16.55mm | 17.11mm | 17.40mm |
| Example three radius of inhibition | 16.72mm | 16.92mm | 17.20mm |
| Example four radius of inhibition | 16.82mm | 17.13mm | 17.36mm |
| Example five radius of inhibition | 16.57mm | 17.25mm | 16.98mm |
In conclusion, the fermentation product obtained by the invention has strong killing and inhibiting effects on vibrio alginolyticus, vibrio anguillarum and vibrio parahaemolyticus of vibrio.
The invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made in the embodiments without departing from the principles and spirit of the invention, and the scope of the invention is to be accorded the broadest interpretation so as to encompass all such modifications and alterations as is within the broadest scope of the claims set forth herein.
Claims (5)
1. A composite probiotic fermentation product for preventing and treating aquatic vibriosis is characterized in that: the fermentation formula of the fermentation product comprises the following components in percentage by mass: 6-12% of scutellaria baicalensis, 3-10% of eucalyptus leaves, 5-7% of dendrobium officinale, 2-4% of bacillus subtilis liquid, 2-4% of candida utilis liquid, 15-20% of corn flour, 15-20% of soybean meal powder, 5-8% of wheat bran, 5-8% of brown sugar and 15-20% of water; the fermentation conditions of the fermentation product are as follows: fermenting at 25-28 deg.C and 50-100r/min for 8-12 days under initial pH of 5-6.
2. The complex probiotic fermentation product for controlling vibriosis in aquatic products according to claim 1, wherein: the fermentation formula of the fermentation product comprises the following components in percentage by mass: 10% of scutellaria baicalensis, 10% of eucalyptus leaves, 7% of dendrobium officinale, 3% of bacillus subtilis liquid, 3% of candida utilis liquid, 17% of corn flour, 20% of soybean meal, 5% of wheat bran, 5% of brown sugar and 20% of water; the fermentation conditions of the fermentation product are as follows: fermenting for 12 days under the conditions of initial pH value of 6, temperature of 25 ℃ and rotation speed of 100 r/min.
3. The complex probiotic fermentation product for controlling vibriosis in aquatic products according to claim 1, wherein: the fermentation formula of the fermentation product comprises the following components in percentage by mass: 12% of scutellaria baicalensis, 8% of eucalyptus leaves, 6% of dendrobium officinale, 4% of bacillus subtilis liquid, 4% of candida utilis liquid, 16% of corn flour, 20% of soybean meal, 5% of wheat bran, 5% of brown sugar and 20% of water; the fermentation conditions of the fermentation product are as follows: fermenting at 28 deg.C and 100r/min for 10 days under initial pH of 5.5.
4. The complex probiotic fermentation product for preventing and treating vibriosis in aquatic products according to any one of claims 1 or 3, wherein the preparation method of the bacillus subtilis liquid comprises the following steps:
uniformly mixing 10g of peptone, 3g of beef extract, 10ml of glucose, 5g of sodium chloride, 10ml of 3.08% magnesium sulfate and 1L of water, and sterilizing for 20 minutes at the temperature of 121 ℃ under the pressure of 0.15MPa by using high-pressure steam to obtain a liquid culture medium for later use;
② inoculating the screened and purified Bacillus subtilis strain to liquid culture medium according to 9% of inoculation amount, performing propagation, performing static culture at 37 deg.C for 24 hr, and adjusting effective viable count to 1x109cfu/ml to obtain the bacillus subtilis liquid.
5. The composite probiotic fermented product for preventing and treating vibriosis in aquatic products according to any one of claims 1 or 3, wherein the candida utilis bacterial liquid is prepared by the following steps:
uniformly mixing 10g of yeast extract powder, 20g of peptone and 20g of glucose, then using distilled water to fix the volume to 1L, and sterilizing for 20 minutes by high-pressure steam at the temperature of 121 ℃ under the pressure of 0.15MPa to obtain a liquid culture medium for later use;
② inoculating the screened and purified Candida utilis strain to liquid culture medium according to 9% of inoculation amount for propagation, performing static culture at 37 ℃ for 24 hours, and adjusting the effective viable count to 1x109cfu/ml to obtain candida utilis bacterial liquid.
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| Publication number | Priority date | Publication date | Assignee | Title |
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