CN111920861A - Bacteriostatic composition of pelteobagrus fulvidraco-derived citrobacter freundii - Google Patents

Bacteriostatic composition of pelteobagrus fulvidraco-derived citrobacter freundii Download PDF

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CN111920861A
CN111920861A CN202010841235.9A CN202010841235A CN111920861A CN 111920861 A CN111920861 A CN 111920861A CN 202010841235 A CN202010841235 A CN 202010841235A CN 111920861 A CN111920861 A CN 111920861A
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derived
bacteriostatic
citrobacter freundii
pelteobagrus fulvidraco
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朱成科
黄莉萍
杨成年
阳龙江
韩璐璐
唐征县
彭小倩
吕光俊
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Southwest University
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Abstract

The invention provides a bacteriostatic composition of Pelteobagrus fulvidraco-derived Citrobacter freundii, which is prepared from the following raw materials in parts by weight: 10-20 parts of traditional Chinese medicine extract, 1-10 parts of bacteriostatic component, 5-10 parts of active component and 1-5 parts of dimethyl sulfoxide. The bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii comprises traditional Chinese medicine components, active components, a bacteriostatic component and dimethyl sulfoxide, wherein the combination of the traditional Chinese medicine components, the dimethyl sulfoxide and the bacteriostatic component has a remarkable bacteriostatic effect on the citrobacter freundii, and the active components increase the stability of other components and improve the bacteriostatic efficiency.

Description

Bacteriostatic composition of pelteobagrus fulvidraco-derived citrobacter freundii
Technical Field
The invention belongs to the field of aquaculture, and particularly relates to a bacteriostatic composition of Pelteobagrus fulvidraco-derived Citrobacter freundii.
Background
The yellow catfish is a common benthic omnivorous fish, and rivers and lakes are distributed with the yellow catfish. The yellow catfish is delicious, high in nutrient meat rate and less in bone spurs, has a medicinal nourishing effect, is one of important economic fishes in China, has the yield of about 50 million tons per year in China, and has the yield of more than 10 hundred million yuan per year. In recent years, the yellow catfish becomes an animal protein nutrition which is loved by consumers, the domestic market is always in an increasing state, and the foreign market has small fluctuation but the price is always high. Therefore, the yellow catfish becomes a foreign exchange variety.
The pelteobagrus fulvidraco is an economic fish widely distributed in fresh water in China, is fond of inhabiting in hard-bottom shallow lakes and rivers, generally moves at the bottom of the water body, gradually floats upwards for foraging, and is a common benthic omnivorous fish. The pelteobagrus fulvidraco lives on the bottom of the river, and prefers to live on a shoal rich in substrate and sludge. The pelteobagrus fulvidraco can breed in the natural condition early in sexual maturity time and has high breeding rate. Sex dimorphism appears on growth, and the spermary matures later than the egg, the male grows faster than the female, and the physique is larger.
The yellow catfish is a meat-preference fish and is generally foraged at night. The food comprises small fish and shrimp food, including small fish, shrimp, various aquatic animals, terrestrial insects, small mollusks and other water-free vertebrates, and the like, and sometimes also can prey on small fishes. The feeding habits of the pelteobagrus fulvidraco are greatly different due to different individual specifications. When the larval fish just comes out of the pellicular membrane, zooplankton such as cladocera and copepods are ingested. When the body length is about 6cm, the feeding of Chironomus larvas, copepods and the like is started. When the body length is more than 9cm, the food mainly eats shrimp, coleoptera larva, snail plant root hair and detritus humus, artificial feed and the like. The change of seasons and environment can also influence the feeding property of the pelteobagrus fulvidraco, for example, the pelteobagrus fulvidraco can often swallow the eggs of other fish in spring and summer, and the situation that the pelteobagrus fulvidraco catches more small and medium fish in autumn and winter, and the benthonic animals can be gradually reduced.
The serious bacterial diseases of the yellow catfish are reported to mainly comprise septicemia, ascites red head body surface ulcer and the like, and the original bacteria mainly comprise aeromonas hydrophila, aeromonas sobria, edwardsiella catarrhalis, edwardsiella tarda, bacillus cereus, vibrio mimicus, ascharomycosis simplicium and the like. And the citrobacter freundii has not been reported and is one of the most serious pathogenic bacteria harming the pelteobagrus fulvidraco in the Chongqing recent years.
In recent years, in Chongqing multi-region pelteobagrus fulvidraco, symptoms such as body ulceration, increased gill mucus, larger ulceration spots on tail fins and tail handles, red and swollen anus, liver khaki and the like appear; the dissected diseased pelteobagrus fulvidraco finds that the gill bleeding erosion has ciliate bulges; swelling and congestion of liver, congestion of intestinal tract, decreased elasticity, and red swelling, congestion and erosion of intestinal tract inner wall. Determined by detection analysis to be Citrobacter freundii.
Disclosure of Invention
In view of the above, the invention aims to overcome the defects in the prior art and provides a bacteriostatic composition of a pseudobagrus fulvidraco source citrobacter freundii.
In order to achieve the purpose, the technical scheme of the invention is realized as follows:
the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii is prepared from the following raw materials in parts by weight:
Figure BDA0002641528040000021
Figure BDA0002641528040000031
preferably, the composition is prepared from the following raw materials in parts by weight:
Figure BDA0002641528040000032
further, the traditional Chinese medicine extract is prepared from the following raw materials in parts by weight: 1-5 parts of fructus chebulae immaturus, 5-10 parts of cortex fraxini, 5-10 parts of pawpaw, 5-10 parts of dark plum fruit, 10-20 parts of gallnut, 10-20 parts of pomegranate rind, 10-20 parts of Chinese pulsatilla root and 10-20 parts of yellow layer.
Preferably, the traditional Chinese medicine extract is prepared from the following raw materials in parts by weight: 3-5 parts of fructus chebulae immaturus, 8-10 parts of cortex fraxini, 8-10 parts of pawpaw, 8-10 parts of dark plum fruit, 15-20 parts of gallnut, 15-20 parts of pomegranate rind, 15-20 parts of Chinese pulsatilla root and 15-20 parts of radix scutellariae. Further, the bacteriostatic component is at least one of chitosan, isobutyl chitosan or chitosan acetylsalicylate.
Further, the active ingredient is hydroxypropyl-beta-cyclodextrin.
The preparation method of the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii comprises the following steps:
(1) pulverizing fructus Chebulae, cortex Fraxini, fructus Chaenomelis, mume fructus, Galla chinensis, pericarpium Granati, radix Pulsatillae and Scutellariae radix respectively, sieving, oven drying, and mixing to obtain Chinese medicinal powder;
(2) adding ethanol into the Chinese medicinal powder, heating in water bath at 60-80 deg.C, reflux extracting, filtering the obtained extractive solution, rotary steaming, concentrating to obtain medicinal storage solution, adding water into the medicinal storage solution, and mixing to obtain mother solution to be tested;
(3) dissolving the bacteriostatic component in water, mixing uniformly and then adding the mixture into the mother solution to be tested;
(4) dissolving the active ingredients in water, mixing uniformly and then adding the mixture into the mother solution to be tested;
(5) and dissolving the dimethyl sulfoxide in water, uniformly mixing, adding the mixture into the mother solution to be tested, and uniformly mixing to obtain the bacteriostatic composition of the Pelteobagrus fulvidraco-derived Citrobacter freundii.
Further, the temperature of the drying step in the step (1) is 35-50 ℃, and the screen mesh of the sieving step is 80-mesh.
Further, the material-liquid ratio of the traditional Chinese medicine powder and the ethanol in the step (2) is 1: 10-15; the material-liquid ratio of the medicine storage liquid to water is 1: 5-10; the time of the reflux step is 1 to 3 hours; the rotating speed of the rotary evaporation step is 100-; the volume of the drug storage solution is one third of the volume of the added ethanol.
Further, the feed-liquid ratio of the bacteriostatic component to water in the step (3) is 1: 5-10; the feed-liquid ratio of the active ingredients to the water in the step (4) is 1: 5-10; the material-liquid ratio of the dimethyl sulfoxide to the water in the step (5) is 1: 100-200.
Compared with the prior art, the invention has the following advantages:
the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii comprises traditional Chinese medicine components, active components, a bacteriostatic component and dimethyl sulfoxide, wherein the combination of the traditional Chinese medicine components, the dimethyl sulfoxide and the bacteriostatic component has a remarkable bacteriostatic effect on the citrobacter freundii, and the active components increase the stability of other components and improve the bacteriostatic efficiency.
The bacteriostatic composition of the Pelteobagrus fulvidraco-derived Citrobacter freundii has a remarkable synergistic effect of Chinese and western olive, ash bark, pawpaw and dark plum, the pawpaw and Chinese gall have a synergistic effect, and the pomegranate bark and the Chinese pulsatilla root have an additive effect.
Detailed Description
Unless defined otherwise, technical terms used in the following examples have the same meanings as commonly understood by one of ordinary skill in the art to which the present invention belongs. The test reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the experimental methods are conventional methods unless otherwise specified.
The invention adopts the citric acid bacillus freundii (PF01) separated from intestinal tracts of the yellow catfish to carry out experiments. The optimal growth temperature of the Citrobacter freundii is about 30 ℃ generally, and the growth of bacteria can be influenced by overhigh or low temperature. If the temperature is lower than 10 ℃ or higher than 35 ℃, hyphae grow slowly. The temperature suitable for the germination of the molecular spores is 28-32 ℃, the molecular spore device can produce spores in four seasons, and the germination and the immersion of the spores are carried out in water for about 8-12 hours.
The components of the invention are all commercial products.
The present invention will be described in detail with reference to examples.
Example 1
The bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii is prepared from the following raw materials in parts by weight: 12 parts of traditional Chinese medicine extract, 5 parts of isobutyl chitosan, 8 parts of hydroxypropyl-beta-cyclodextrin and 2 parts of dimethyl sulfoxide.
The traditional Chinese medicine extract is prepared from the following raw materials in parts by weight: 5 parts of fructus chebulae immaturus, 10 parts of cortex fraxini, 10 parts of pawpaw, 10 parts of dark plum fruit, 20 parts of gallnut, 20 parts of pomegranate rind, 20 parts of Chinese pulsatilla root and 20 parts of radix scutellariae.
The preparation method of the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii comprises the following steps:
(1) pulverizing fructus Chebulae, cortex Fraxini, fructus Chaenomelis, mume fructus, Galla chinensis, pericarpium Granati, and radix Pulsatillae respectively, sieving with 80 mesh sieve, oven drying at 40 deg.C, and mixing to obtain Chinese medicinal powder;
(2) adding 10 times of ethanol into the traditional Chinese medicine powder, heating in a water bath at 60 ℃, carrying out reflux extraction for 2 hours, filtering the obtained extracting solution, carrying out rotary evaporation at 150r/min, concentrating to obtain a medicine storage solution after the volume of the extracting solution is one third of the original ethanol addition amount, then adding 5 times of water into the medicine storage solution, and uniformly mixing to obtain a mother solution to be tested;
(3) dissolving the bacteriostatic component in 5 times of water, uniformly mixing, and adding into the mother solution to be tested;
(4) dissolving the active ingredients in 5 times of water, uniformly mixing, and adding into the mother solution to be tested;
(5) dissolving the dimethyl sulfoxide in 100 times of water, uniformly mixing, adding into the mother solution to be tested, and uniformly mixing to obtain the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii.
Example 2
The bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii is prepared from the following raw materials in parts by weight: 20 parts of traditional Chinese medicine extract, 5 parts of isobutyl chitosan, 8 parts of hydroxypropyl-beta-cyclodextrin and 2 parts of dimethyl sulfoxide.
The traditional Chinese medicine extract is prepared from the following raw materials in parts by weight: 5 parts of fructus chebulae immaturus, 10 parts of cortex fraxini, 10 parts of pawpaw, 10 parts of dark plum fruit, 20 parts of gallnut, 20 parts of pomegranate rind, 20 parts of Chinese pulsatilla root and 20 parts of radix scutellariae.
The preparation method of the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii comprises the following steps:
(1) pulverizing fructus Chebulae, cortex Fraxini, fructus Chaenomelis, mume fructus, Galla chinensis, pericarpium Granati, and radix Pulsatillae respectively, sieving with 80 mesh sieve, oven drying at 40 deg.C, and mixing to obtain Chinese medicinal powder;
(2) adding 10 times of ethanol into the traditional Chinese medicine powder, heating in a water bath at 60 ℃, carrying out reflux extraction for 2 hours, filtering the obtained extracting solution, carrying out rotary evaporation at 150r/min, concentrating to obtain a medicine storage solution after the volume of the extracting solution is one third of the original ethanol addition amount, then adding 5 times of water into the medicine storage solution, and uniformly mixing to obtain a mother solution to be tested;
(3) dissolving the bacteriostatic component in 5 times of water, uniformly mixing, and adding into the mother solution to be tested;
(4) dissolving the active ingredients in 5 times of water, uniformly mixing, and adding into the mother solution to be tested;
(5) dissolving the dimethyl sulfoxide in 100 times of water, uniformly mixing, adding into the mother solution to be tested, and uniformly mixing to obtain the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii.
Example 3
The bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii is prepared from the following raw materials in parts by weight: 12 parts of traditional Chinese medicine extract, 10 parts of isobutyl chitosan, 8 parts of hydroxypropyl-beta-cyclodextrin and 2 parts of dimethyl sulfoxide.
The traditional Chinese medicine extract is prepared from the following raw materials in parts by weight: 5 parts of fructus chebulae immaturus, 10 parts of cortex fraxini, 10 parts of pawpaw, 10 parts of dark plum fruit, 20 parts of gallnut, 20 parts of pomegranate rind, 20 parts of Chinese pulsatilla root and 20 parts of radix scutellariae.
The preparation method of the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii comprises the following steps:
(1) pulverizing fructus Chebulae, cortex Fraxini, fructus Chaenomelis, mume fructus, Galla chinensis, pericarpium Granati, and radix Pulsatillae respectively, sieving with 80 mesh sieve, oven drying at 40 deg.C, and mixing to obtain Chinese medicinal powder;
(2) adding 10 times of ethanol into the traditional Chinese medicine powder, heating in a water bath at 60 ℃, carrying out reflux extraction for 2 hours, filtering the obtained extracting solution, carrying out rotary evaporation at 150r/min, concentrating to obtain a medicine storage solution after the volume of the extracting solution is one third of the original ethanol addition amount, then adding 5 times of water into the medicine storage solution, and uniformly mixing to obtain a mother solution to be tested;
(3) dissolving the bacteriostatic component in 5 times of water, uniformly mixing, and adding into the mother solution to be tested;
(4) dissolving the active ingredients in 5 times of water, uniformly mixing, and adding into the mother solution to be tested;
(5) dissolving the dimethyl sulfoxide in 100 times of water, uniformly mixing, adding into the mother solution to be tested, and uniformly mixing to obtain the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii.
Example 4
The bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii is prepared from the following raw materials in parts by weight: 12 parts of traditional Chinese medicine extract, 5 parts of isobutyl chitosan, 8 parts of hydroxypropyl-beta-cyclodextrin and 5 parts of dimethyl sulfoxide.
The traditional Chinese medicine extract is prepared from the following raw materials in parts by weight: 5 parts of fructus chebulae immaturus, 10 parts of cortex fraxini, 10 parts of pawpaw, 10 parts of dark plum fruit, 20 parts of gallnut, 20 parts of pomegranate rind, 20 parts of Chinese pulsatilla root and 20 parts of radix scutellariae.
The preparation method of the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii comprises the following steps:
(1) pulverizing fructus Chebulae, cortex Fraxini, fructus Chaenomelis, mume fructus, Galla chinensis, pericarpium Granati, and radix Pulsatillae respectively, sieving with 80 mesh sieve, oven drying at 40 deg.C, and mixing to obtain Chinese medicinal powder;
(2) adding 10 times of ethanol into the traditional Chinese medicine powder, heating in a water bath at 60 ℃, carrying out reflux extraction for 2 hours, filtering the obtained extracting solution, carrying out rotary evaporation at 150r/min, concentrating to obtain a medicine storage solution after the volume of the extracting solution is one third of the original ethanol addition amount, then adding 5 times of water into the medicine storage solution, and uniformly mixing to obtain a mother solution to be tested;
(3) dissolving the bacteriostatic component in 5 times of water, uniformly mixing, and adding into the mother solution to be tested;
(4) dissolving the active ingredients in 5 times of water, uniformly mixing, and adding into the mother solution to be tested;
(5) dissolving the dimethyl sulfoxide in 100 times of water, uniformly mixing, adding into the mother solution to be tested, and uniformly mixing to obtain the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii.
Comparative example 1
The bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii is prepared from the following raw materials in parts by weight: 5 parts of fructus chebulae immaturus, 10 parts of cortex fraxini, 10 parts of pawpaw, 10 parts of dark plum fruit, 20 parts of gallnut, 20 parts of pomegranate rind, 20 parts of Chinese pulsatilla root and 20 parts of radix scutellariae.
The preparation method of the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii comprises the following steps:
(1) pulverizing fructus Chebulae, cortex Fraxini, fructus Chaenomelis, mume fructus, Galla chinensis, pericarpium Granati, and radix Pulsatillae respectively, sieving with 80 mesh sieve, oven drying at 40 deg.C, and mixing to obtain Chinese medicinal powder;
(2) adding 10 times of ethanol into the traditional Chinese medicine powder, heating in a water bath at 60 ℃, carrying out reflux extraction for 2 hours, filtering the obtained extracting solution, carrying out rotary evaporation at 150r/min, concentrating to obtain a medicine storage solution after the volume of the extracting solution is one third of the original ethanol addition amount, then adding 5 times of water into the medicine storage solution, and uniformly mixing to obtain the bacteriostatic composition of the pseudobagrus fulvidraco-derived citrobacter freundii.
Comparative example 2
The bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii is prepared from the following raw materials in parts by weight: 12 parts of traditional Chinese medicine extract, 5 parts of isobutyl chitosan and 2 parts of dimethyl sulfoxide.
The traditional Chinese medicine extract is prepared from the following raw materials in parts by weight: 5 parts of fructus chebulae immaturus, 10 parts of cortex fraxini, 10 parts of pawpaw, 10 parts of dark plum fruit, 20 parts of gallnut, 20 parts of pomegranate rind, 20 parts of Chinese pulsatilla root and 20 parts of radix scutellariae.
The preparation method of the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii comprises the following steps:
(1) pulverizing fructus Chebulae, cortex Fraxini, fructus Chaenomelis, mume fructus, Galla chinensis, pericarpium Granati, and radix Pulsatillae respectively, sieving with 80 mesh sieve, oven drying at 40 deg.C, and mixing to obtain Chinese medicinal powder;
(2) adding 10 times of ethanol into the traditional Chinese medicine powder, heating in a water bath at 60 ℃, carrying out reflux extraction for 2 hours, filtering the obtained extracting solution, carrying out rotary evaporation at 150r/min, concentrating to obtain a medicine storage solution after the volume of the extracting solution is one third of the original ethanol addition amount, then adding 5 times of water into the medicine storage solution, and uniformly mixing to obtain a mother solution to be tested;
(3) dissolving the bacteriostatic component in 5 times of water, uniformly mixing, and adding into the mother solution to be tested;
(4) dissolving the dimethyl sulfoxide in 100 times of water, uniformly mixing, adding into the mother solution to be tested, and uniformly mixing to obtain the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii.
12 Chinese herbal medicines (shown in table 1) are selected, 66 duplex compounds (FC01-FC66) are designed by a chessboard method and used for in vitro bacteriostasis experiments, and the results are shown in table 2. The minimum bacteriostatic concentration of most Chinese herbal medicines in the combined compound is lower than that of the Chinese herbal medicines when the Chinese herbal medicines are used singly. The MIC of the 4 Chinese herbal medicine compounds of the fructus chebulae immaturus, the ash bark, the pawpaw and the dark plum is obviously reduced, the drug effect is obviously improved, and the synergistic effect is obvious. The two Chinese herbal medicines in the compound of pawpaw, Chinese gall and the like have synergistic effect. The two Chinese herbal medicines in the compound of pomegranate rind and Chinese pulsatilla root are added; two Chinese herbal medicines in the compound medicine of the myrobalan, the Chinese pulsatilla root and the like have no relevant effect.
Table 1 for short Chinese herbal medicine in compound prescription
Figure BDA0002641528040000121
TABLE 2 double-compound Chinese herbal medicine effect on inhibiting Citrobacter freundii
Figure BDA0002641528040000122
Note: e: evaluation: o: remarkable synergy; s: synergy effect; t: addition; i: an unrelated effect;
the results in Table 2 show that reasonable compatibility has a certain synergistic effect on the drug effects of the Chinese herbal medicines, and the use concentration of each Chinese herbal medicine in the compound is reduced by at least 50%, which indicates that the reasonable combination of the Chinese herbal medicines can reduce the use concentration of the medicines on the basis of ensuring the drug effects, reduce the medicine residues and provide a new idea for the development of fishery antibacterial medicines.
The Chinese herbal medicine compound generates synergistic effect and is related to the effective bacteriostatic component of the traditional Chinese medicine. The Chinese herbal medicine compound generates synergistic effect and is related to the effective bacteriostatic component of the traditional Chinese medicine. Different traditional Chinese medicine components and different bacteriostatic mechanisms are combined, different action ways and Chinese herbal medicines with different target sites can attack the cells of pathogenic bacteria in multiple ways and multiple sites, and the bactericidal capacity of the medicine is greatly improved.
The strain was inoculated on LB solid medium, and then the compositions obtained in examples 1-4 and comparative examples 1-2 were added at different dilution ratios, and after culturing under aseptic conditions, the results were observed as shown in Table 3.
TABLE 3 results of the experiment
Dilution factor Example 1 Example 2 Example 3 Example 4 Comparative example 1 Comparative example 2
0 - - - + + +
5 + + + + ++ +
10 + + ++ ++ ++ +++
20 ++ + +++ +++ ++++ +++
100 +++ + +++ +++ ++++ ++++
1000 +++ +++ +++ ++++ ++++ ++++
5000 +++ +++ ++++ ++++ ++++ ++++
10000 +++ +++ ++++ ++++ ++++ ++++
Positive control ++++ ++++ ++++ ++++ ++++ ++++
Negative control - - - - - -
Note: "-" indicates no hyphal growth observed visually. "+" indicates 1/4 where the intensity was less than the diameter of the wells, "+ +" indicates 1/4-/2 where the length of the hyphae was the diameter of the wells, "+++" indicates 1/2-/4 where the length of the hyphae was the diameter of the wells, and "+++++" indicates that the length of the hyphae was 3/4 or more of the diameter of the wells.
As shown in table 3, the compositions obtained in examples 1 to 3 all have significant bacteriostatic effects on citrobacter freundii, wherein the composition obtained in example 2 has the most significant effect, which indicates that the traditional Chinese medicine components have significant inhibitory effects on citrobacter freundii, and the increase in the addition amount of the isobutylchitosan and the dimethyl sulfoxide does not bring about more excellent bacteriostatic effects, indicating that the concentration of the isobutylchitosan and the dimethyl sulfoxide should be at a lower level, which is beneficial to the bacteriostatic action of the composition, and the traditional Chinese medicine components are only added in comparative example 1, so that the bacteriostatic effects are greatly reduced compared with those in examples 1 to 4, and the bacteriostatic effects of the components cannot be fully exerted because no active components are added in comparative example 2.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Claims (10)

1. A bacteriostatic composition of Pelteobagrus fulvidraco-derived Citrobacter freundii is characterized in that: the composition is prepared from the following raw materials in parts by weight:
Figure FDA0002641528030000011
2. the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii according to claim 1, which is characterized in that: the composition is prepared from the following raw materials in parts by weight:
Figure FDA0002641528030000012
3. the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii according to claim 2, which is characterized in that: the traditional Chinese medicine extract is prepared from the following raw materials in parts by weight: 1-5 parts of fructus chebulae immaturus, 5-10 parts of cortex fraxini, 5-10 parts of pawpaw, 5-10 parts of dark plum fruit, 10-20 parts of gallnut, 10-20 parts of pomegranate rind, 10-20 parts of Chinese pulsatilla root and 10-20 parts of yellow layer.
4. The bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii according to claim 3, which is characterized in that: the traditional Chinese medicine extract is prepared from the following raw materials in parts by weight: 3-5 parts of fructus chebulae immaturus, 8-10 parts of cortex fraxini, 8-10 parts of pawpaw, 8-10 parts of dark plum fruit, 15-20 parts of gallnut, 15-20 parts of pomegranate rind, 15-20 parts of Chinese pulsatilla root and 15-20 parts of radix scutellariae.
5. The bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii according to claim 4, which is characterized in that: the bacteriostatic component is at least one of chitosan, isobutyl chitosan or chitosan acetylsalicylate.
6. The bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii according to claim 5, wherein the bacteriostatic composition comprises the following components in percentage by weight: the active ingredient is hydroxypropyl-beta-cyclodextrin.
7. The preparation method of the bacteriostatic composition of the pelteobagrus fulvidraco-derived citrobacter freundii according to claim 6, which is characterized by comprising the following steps: the method comprises the following steps:
(1) pulverizing fructus Chebulae, cortex Fraxini, fructus Chaenomelis, mume fructus, Galla chinensis, pericarpium Granati, radix Pulsatillae and Scutellariae radix respectively, sieving, oven drying, and mixing to obtain Chinese medicinal powder;
(2) adding ethanol into the Chinese medicinal powder, heating in water bath at 60-80 deg.C, reflux extracting, filtering the obtained extractive solution, rotary steaming, concentrating to obtain medicinal storage solution, adding water into the medicinal storage solution, and mixing to obtain mother solution to be tested;
(3) dissolving the bacteriostatic component in water, mixing uniformly and then adding the mixture into the mother solution to be tested;
(4) dissolving the active ingredients in water, mixing uniformly and then adding the mixture into the mother solution to be tested;
(5) and dissolving the dimethyl sulfoxide in water, uniformly mixing, adding the mixture into the mother solution to be tested, and uniformly mixing to obtain the bacteriostatic composition of the Pelteobagrus fulvidraco-derived Citrobacter freundii.
8. The preparation method of the bacteriostatic composition of the Pelteobagrus fulvidraco-derived Citrobacter freundii according to claim 7, characterized by comprising the following steps: the temperature of the drying step in the step (1) is 35-50 ℃, and the screen of the sieving step is an 80-mesh screen.
9. The preparation method of the bacteriostatic composition of the Pelteobagrus fulvidraco-derived Citrobacter freundii according to claim 7, characterized by comprising the following steps: the material-liquid ratio of the traditional Chinese medicine powder to the ethanol in the step (2) is 1: 10-15; the material-liquid ratio of the medicine storage liquid to water is 1: 5-10; the time of the reflux step is 1 to 3 hours; the rotating speed of the rotary evaporation step is 100-; the volume of the drug storage solution is one third of the volume of the added ethanol.
10. The preparation method of the bacteriostatic composition of the Pelteobagrus fulvidraco-derived Citrobacter freundii according to claim 7, characterized by comprising the following steps: the feed-liquid ratio of the bacteriostatic component to water in the step (3) is 1: 5-10; the feed-liquid ratio of the active ingredients to the water in the step (4) is 1: 5-10; the material-liquid ratio of the dimethyl sulfoxide to the water in the step (5) is 1: 100-200.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112691184A (en) * 2020-12-31 2021-04-23 西藏自治区农牧科学院水产科学研究所 Bacteriostatic composition for inhibiting saprolegnia and preparation method thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
凌空等: "十六种中草药及其复方对大鲵致病性豚鼠气单胞菌的体外抑制作用", 《淡水渔业》 *
金娟: "两种大鲵病原菌的分离鉴定及其理化特性研究", 《中国优秀博硕士学位论文全文数据库(硕士) 农业科技辑》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112691184A (en) * 2020-12-31 2021-04-23 西藏自治区农牧科学院水产科学研究所 Bacteriostatic composition for inhibiting saprolegnia and preparation method thereof

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