KR20130013965A - Anti-fungal and anti-bacterial compositions against fish pathogenic bacteria - Google Patents
Anti-fungal and anti-bacterial compositions against fish pathogenic bacteria Download PDFInfo
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- KR20130013965A KR20130013965A KR1020110075926A KR20110075926A KR20130013965A KR 20130013965 A KR20130013965 A KR 20130013965A KR 1020110075926 A KR1020110075926 A KR 1020110075926A KR 20110075926 A KR20110075926 A KR 20110075926A KR 20130013965 A KR20130013965 A KR 20130013965A
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- extract
- antifungal
- fish
- rhubarb
- bokbunja
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
- A01K61/13—Prevention or treatment of fish diseases
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/489—Sophora, e.g. necklacepod or mamani
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
- A61K36/708—Rheum (rhubarb)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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Abstract
The present invention relates to an antifungal and antimicrobial composition against fish bacterial pathogens, and more particularly, to an antifungal and antimicrobial composition of fish comprising one or more extracts selected from the group consisting of red ginseng, rhubarb and bokbunja as an active ingredient. .
According to the present invention, using an antifungal and antimicrobial composition comprising a ginseng extract, a rhubarb extract, a bokbunja extract, or a mixture thereof, infection of fish of a strain which is a bacterial disease agent causing enormous economic loss to freshwater and seawater culture in Korea It can be used to prevent and treat. Natural ingredients such as red ginseng, rhubarb and bokbunja extracts can ultimately replace the chemicals currently in use and provide more environmentally friendly, high value added natural antifungal and antimicrobial agents. Can be.
Description
The present invention relates to an antifungal and antimicrobial composition against fish bacterial pathogens, and more particularly, to an antifungal and antimicrobial composition of fish comprising at least one extract selected from the group consisting of red ginseng, rhubarb and bokbunja as an active ingredient. .
Aquatic fungi are mycelia that develop and develop spores only on dead cells or tissues, but have been reported to be very difficult to cure due to infinite reproduction by penetrating deep into healthy cells or three lobes after infection. There is also a high probability of developing aquatic fungal infections when there is trauma, when long-term or deteriorated feeds are fed, when nutrition is poor after wintering, and when the temperature changes in early spring are severe. In particular, because fertilized eggs have no movement and the fertilized eggs and the fertilized eggs are in contact with each other, when infected by aquatic fungi caused by the eggs, their spread is very fast and also occurs during spawning and incubation periods. Inflicts damage. Malachite Green, which has been used for aquatic fungal infections in freshwater fish, has been used worldwide since 1949. It has been banned, and its use has been banned entirely since 2003 in Korea and Japan. As malachite green has been identified as a carcinogenic substance and its use is banned, formalin, bronopol, humic acid, chlorine dioxide, and hydrogen peroxide are mainly used as alternative chemicals. The most commonly used formalin (formalin) is an aqueous solution of formaldehyde. Formaldehyde is a flammable colorless gas with an irritating odor. Soluble in water, 37% aqueous solution of formaldehyde is called formalin. Formalin is mainly used as antiseptic and preservative. Formaldehyde is very toxic to the human body, and when a person is exposed to more than 30 ppm of formaldehyde, symptoms begin to develop. Powerful protein coagulation invades skin and mucous membranes, and inhalation with gas can cause pharyngitis or bronchitis. High doses can lead to heart failure and death. If you take 100 mg (0.1 g) per kg of body weight, 50% of people die. This means that if 100 g of a person weighing 70 kg is ingested 7 g each, 50 of them will die. According to a Schenke report written in 1981, exposure to 30 ppm in air for 1 minute can cause symptoms such as memory loss and mental distress. When formaldehyde is disposed of, it is required to be incinerated at high temperature or melted and solidified at high temperature according to the Waste Chemicals Control Act and the Waste Management Act. As such, in the case of the use of chemicals such as formalin, as with malachite green, the possibility of adverse effects on the human body is very high. Therefore, in order to secure the safety of aquatic products, antifungal substances of natural materials are required.
Therefore, the present inventors have made diligent research efforts to overcome the problems of the prior art, when treating ginseng extract, rhubarb extract, bokbunja extract or mixtures thereof, as well as antifungal activity against aquatic fungi, as well as disease in aquaculture fish It was confirmed that the present invention can exhibit effective antimicrobial activity against the resulting Vibrio strain, Streptococcus strain and Edwardsiela strain, and completed the present invention.
Accordingly, the main object of the present invention is to provide a natural antifungal and antimicrobial composition having an antifungal and antimicrobial effect of inhibiting fish aquatic fungal infection with high safety and efficacy compared to formalin which has been used for aquatic fungal infection.
Another object of the present invention to provide a feed composition for fish feed using the natural antifungal and antimicrobial composition.
Still another object of the present invention is to provide a method for preparing the natural antifungal and antimicrobial composition.
According to an aspect of the present invention, the present invention provides an antifungal and antimicrobial composition of fish comprising at least one extract selected from the group consisting of red ginseng, rhubarb, bokbunja as an active ingredient.
As an active ingredient of the present invention, the ginseng ( Sophora flavescens Aiton) is a perennial herbaceous plant belonging to the legume and grows all over Korea. Korea, China, and Japan all use roots barely stripped of ginseng, and Korea and Japan also use ginseng powder. Ginseng has a peculiar smell, is persistent, and weak. Pharmacological effects have been reported to treat leukopenia, anti-radioactivity, increased coronary blood flow, strengthening heart muscle, lowering blood sugar, anti-tumor, immune function. Major components of ginseng are alkaloids such as d-matrine, d-oxyma trine, d-sophoranol, and flavonoids such as xanthohumol and isoxanthohumol.
Rhubarb is the name given because it is yellow in color, and the effect is very fast, the nature is very cold, the taste is bitter, and there is no poison. Pharmacological effects have been reported to promote colon exercise, lowering action, fever, lowering body temperature, promoting bile secretion, reducing blood clotting time, diuresis, and protecting liver function. Its main ingredients include Emodin, Chrisophamol, Rhein, Aloe-emodin and Glucogallin.
The bokbunja is a medicinal herb made from raw fruit of the Robus bokbunja strawberry ( Rubis coreanus Miquel) (Korea and China), and in Japan it was not modified as a process herbal medicine. Bokbunja has no smell, tastes sour and sweet, and its properties are warm. As a pharmacological effect, antioxidant activity and anti-Helicobacter pylori action have been reported. Appearance is a small cluster of small clusters of lumps, which form a conical or pressed sphere. The outer side is yellowish green or pale brown, the end is rounded, and the center of the calyx is recessed. Its main components include cyanidin-3-sambubioside, cyanidin-3-glucoside, cyanidin-3-xylosylrutinoside, cyanidin-3-rutinoside and pelargonidin-3-rutinoside.
These components are not a simple molecular structure but have a very complex structure, and about 100 other components are expected to have strong therapeutic power and antibacterial properties by interacting with each other. Therefore, when mixing the natural extract of the present invention exhibits excellent antifungal and antibacterial action by the synergistic effect caused by the main components of the extract.
In the antifungal and antimicrobial composition of the fish of the present invention, the composition may include the extract of red ginseng, rhubarb and bokbunja alone, it may exhibit an elevated antifungal and antibacterial activity when the two or more of the extracts are mixed. In particular, it is preferable to include the ginseng extract and rhubarb extract, the ginseng extract and bokbunja extract, or rhubarb extract and bokbunja extract in a 1: 3 to 3: 1 weight ratio.
In the embodiment of the present invention, the anti-fungal and antimicrobial effects of the extracts were treated by treating ginseng, rhubarb and bokbunja extracts alone. As a result, compared with formalin treatment, a chemical used in aquaculture farms, the antibacterial activity was increased by treatment of each extract of the present invention (see FIGS. 2A, 2B and Table 1). The antimicrobial activity was measured for S. It showed antimicrobial activity against iniae , V for Bokbunja extract. anguillarum and S. It showed antimicrobial activity against iniae (see Tables 4-6).
In a preferred embodiment of the present invention, when the weight ratio of the ginseng and rhubarb extract and rhubarb and bokbunja extract is 1: 1, respectively, showed the highest synergistic effect in the antifungal effect of aquatic fungi (see Table 2).
In the embodiment of the present invention, the antifungal effect of the mixture was treated by treating the mixture of the ginseng extract, rhubarb extract and bokbunja extract with 1: 3 to 3: 1 weight ratio of each of the two kinds of extracts. As a result, the same amount of ginseng and rhubarb extract, the same amount of bokbunja and rhubarb extract showed elevated antifungal activity by the mixed treatment.
In addition, the composition may also exhibit an elevated antimicrobial activity when mixed with both ginseng extract, rhubarb extract and bokbunja extract. It is preferable that the mixing ratio of these extracts include ginseng extract, rhubarb extract, and bokbunja extract in a ratio of 1 to 3: 1 to 3: 1 to 3 by weight. More preferably, the ginseng extract, rhubarb extract and bokbunja extract is preferably mixed in a 1: 1: 1 weight ratio.
Red ginseng, rhubarb and bokbunja extract used as an active ingredient of the present invention can be obtained using an extraction method known in the art. For example, the red ginseng, rhubarb, bokbunja or a pulverized product thereof may be extracted with at least one solvent selected from the group consisting of water, ethanol, isopropyl alcohol, methanol, propanol, butanol, glycerin, butylene glycol. The extraction method may be a method of immersing red ginseng, rhubarb or bokbunja pulverized in a solvent for 3 to 5 hours at 70 to 90 ℃.
In the present invention, the composition has an antifungal activity against aquatic fungi ( Saprolegnia parasitica ) and antimicrobial activity against fish pathogenic bacteria, including Vibrio strain, Streptococcus strain, Edwardcyela strain, which occupy most of the bacterial fish diseases in the farm. And preferably, Vibrio anguillarum , Streptococcus iniae ), Edwardsiella tarda ( edwardsiella tarda ) is characterized by having an antimicrobial activity.
In the present invention, the composition may be administered to an animal for the purpose of preventing and treating a bacterial infection, but is preferably administered for the treatment of a fungal disease and / or a bacterial disease of fish. Examples of the administrable fish are characterized in that selected from the group consisting of halibut, sea bass rock, defense, trout, eel, carp and catfish.
The antifungal and antimicrobial composition of the present invention may be administered directly or indirectly to fish, and the form to be administered may also be used in the form of a salt that is acceptable as an antimicrobial use of these fish. It may also be used alone or in combination with other veterinary active compounds, as well as in suitable sets.
The antifungal and antimicrobial composition of the present invention may further include appropriate excipients and diluents commonly used in the preparation of the antifungal and antimicrobial compositions.
Examples of the excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose. , Polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
In addition, the antifungal and antimicrobial composition may further include a filler, an anticoagulant, a lubricant, a humectant, a spice, an emulsifier, a preservative, and the like. Formulations may be formulated using methods well known in the art to provide release, the formulations may be in powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, suppositories, Sterile injectable solutions, sterile external preparations, and the like.]
The antifungal and antimicrobial composition according to the present invention may vary depending on the age, sex and weight of the fish, but may be administered once to several times in an amount of 0.01 to 100 mg / kg. The dosage may also be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. Accordingly, the dosage is not limited in any way to the scope of the present invention.
According to another aspect of the present invention, the present invention provides a fish feed composition comprising the antifungal and antimicrobial composition of the fish as an active ingredient.
According to another aspect of the invention, the present invention comprises the steps of a) extracting ginseng, rhubarb or bokbunja with an organic solvent; b) mixing the one or more extracts; And c) mixing the mixture with a veterinary acceptable excipient and diluent to formulate the composition.
As described above, according to the present invention, by using an antifungal and antimicrobial composition comprising a ginseng extract, rhubarb extract, bokbunja extract, or a mixture thereof, a strain of bacterial disease source causing enormous economic loss to freshwater and seawater culture in Korea. It can be used to prevent and treat infections in fish.
Natural ingredients such as red ginseng, rhubarb and bokbunja extracts can ultimately replace the chemicals currently in use and provide more environmentally friendly, high value added natural antifungal and antimicrobial agents. Can be.
Figure 1 shows an antifungal experimental schematic.
Figure 2a is the result of measuring the antifungal effect according to formalin treatment (D inh = size of the low zone formed by formalin (cm)), Figure 2b shows the result of measuring the antifungal effect of the medicinal plant extract treatment of the present invention will be.
Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, and the scope of the present invention is not to be construed as being limited by these examples.
Example 1. Preparation of medicinal plant extracts
After grinding each of red ginseng, rhubarb and bokbunja, each of the above pulverized product was added to 70% methanol, 70% ethanol or water in 10% content, and extracted by soaking at 80 ° C. for 5 hours. . The red ginseng, rhubarb, and bokbunja were extracted from each of the three solvents (70% methanol, 70% ethanol and water), respectively, and thus the total extract samples were nine.
Example 2. Isolation and Identification of Aquatic Fungi
Aquatic fungi were isolated from the larvae of aquaculture farms, and three to five pure separation processes were performed. After aquatic fungi were selected, they were identified through sequencing of the internal transcript spacer (ITS), and finally Saprolegnia. parasitica was used.
Example 3. Antifungal Experiment
For antifungal experiments, formalin (formalin), a chemical used in aquaculture farms, and three medicinal plant extracts according to the present invention (Ginseng, Rhubarb, and Bokbunja extracts) were used as test materials. Specific antifungal experiments are as follows, the experimental schematic is shown in FIG.
A 0.8 mm stainless hole was placed on a GY (Yeast extract, 2.5 g; 10 g / L glucose) plate, and a hole was formed by pouring GY soft medium. Then, 50 μl of 10% medicinal plant extracts were applied to the formed holes, and 1 cm 2 aquatic fungus ( Saprolegnia) in the center of the medium. parasitica ) blocks were raised and incubated at 20 ° C. for 3 days. After incubation was completed, the size of the lowland was measured. The measurement results are shown in FIG. 2.
As shown in Figure 2a, as a result of treatment of aquatic fungi with formalin, the size of the lowland was 2 cm at 100 ppm diluted with formalin (37%), 100 cm at 0.2 ppm, and at 100 ppm for control (instead of formalin). It was found that the growth of aquatic fungi could not be prevented. On the other hand, as shown in Figure 2b and Table 1, when treating the medicinal plant extract according to the present invention was confirmed to significantly inhibit the growth of aquatic fungi, in particular, ginseng, rhubarb, bokbunja extract extracted with 70% methanol In the case of treatment of the low zones of 2.6, 2.95, and 2.9 cm, respectively, it was confirmed that the growth of aquatic fungus more effectively than when formalin was treated with 10,000 ppm.
[Table 1. Antifungal Effects of Medicinal Plant Extract Treatments]
Example 4. Antifungal Synergy Effect of Medicinal Plant Extracts
Through the antifungal experiment of Example 3, it was confirmed that the antifungal effect by mixing high ginseng, rhubarb, bokbunja extract (all 70% methanol solvent extract) showed high efficacy. The mixed extract samples were classified into four types, and were prepared by mixing the same weight ratios of red ginseng + rhubarb extract, red ginseng + bokbunja extract, rhubarb + bokbunja extract and red ginseng + rhubarb + bokbunja extract. Aquatic fungi used in antifungal experiments are the most common onset of aquaculture in Saprolegnia. parasitica was used. The antifungal test method was performed in the same manner as described in Example 3, and 50 μl of 10% mixed extract was applied to each sample. After completion of the final culture, the size of the lowland was measured. The measurement results are shown in Table 2 below.
As a result, it can be seen that the treatment of the mixed extract inhibits the growth of aquatic fungi more efficiently than the treatment of a single medicinal plant extract, and in particular, all three medicinal plant extracts mixed with red ginseng, rhubarb and bokbunja are treated. The lowest antifungal effect was found at 3.4 cm.
[Table 2. Antifungal Effects of Medicinal Plant Mixture Extract Treatments]
Example 5. Having antifungal efficacy Gosam , rhubarb Antimicrobial experiment using fish disease bacteria
As confirmed in Examples 3 and 4, in order to determine whether the extract of red ginseng, rhubarb, and bokbunja having high antifungal efficacy is also effective in antibacterial, antibacterial experiment was performed using the fish disease bacteria described in Table 3 below. At this time, Fish Pathology, Vibrio bacteria, which accounts for most of the bacterial strains used fish diseases (Vibrio anguillarum), Streptococcus strain (Streptococcus iniae ) and Edwardsiella tarda were selected first and used for antimicrobial experiments. The strain of each of these bacteria is shown in Table 3 below.
[Table 3. Fish Bacteria Used in Antimicrobial Experiments]
The antimicrobial test method is as follows. ST medium (NaCl 1.5%, Tryptic Soy Broth 30 g and agar 1.5% / 1 L), which is the medium of each strain, was poured into a plate (petri-dish) and hardened, and then a stainless hole having a diameter of 0.8 cm was hardened. It was placed on a plate. Then, in the ST medium composition, 10 mL of ST agar medium (ST soft-agar medium) containing agar content of 0.75% in the ST medium composition was adjusted to the absorbance (OD 600 ) of each strain to 0.1 100 After addition of μl, the medium containing the strain is poured and solidified onto the medium on which stainless holes are erected. Thereafter, after removing the stainless hole, 50 μL of the medicinal plant extract (Ginseng, rhubarb or bokbunja extract) according to the present invention was added to the resulting hole, followed by culturing for 18 hours in a 27 ° C. incubator, whether the experimental strains were grown around the hole. It was confirmed. If the effect is greater, a clear zone will appear around the hole, and in the case of a sample without antimicrobial activity, the strains grow around the hole so that the low zone is not visible. The diameter size of these lowlands indicates the antimicrobial activity of the extract. The measurement results are shown in Tables 4 to 6 below.
Antimicrobial effect against fish disease bacteria according to the medicinal plant extract of the present invention, 70% ethanol and methanol extract S. It had antimicrobial effect against iniae , and all extracts were treated with S. The antimicrobial activity was about iniae, when processing a bokbunja extract is 70% ethanol and methanol extracts from V. anguillarum and S. It was confirmed that there is an antimicrobial effect on iniae .
[Table 4. Antimicrobial Effects of Ginseng Extract Treatment]
[Table 5. Antibacterial Effect of Rhubarb Extract Treatment]
[Table 6. Antimicrobial Effect According to Bokbunja Extract Treatment]
Claims (9)
b) mixing the one or more extracts; And
c) A method for preparing an antifungal and antimicrobial composition of fish comprising mixing and formulating the mixture with a veterinary acceptable excipient and diluent.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103142711A (en) * | 2013-03-22 | 2013-06-12 | 中山火炬职业技术学院 | Anti-cancer pharmaceutical composition, and preparation method as well as application thereof |
| CN105213500A (en) * | 2015-11-11 | 2016-01-06 | 郑州后羿制药有限公司 | A kind of control fresh-water fishes mycotic Chinese herbal medicine fermented extracted liquid and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103142711A (en) * | 2013-03-22 | 2013-06-12 | 中山火炬职业技术学院 | Anti-cancer pharmaceutical composition, and preparation method as well as application thereof |
| CN105213500A (en) * | 2015-11-11 | 2016-01-06 | 郑州后羿制药有限公司 | A kind of control fresh-water fishes mycotic Chinese herbal medicine fermented extracted liquid and preparation method thereof |
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