CN113599432B

CN113599432B - Traditional Chinese medicine for preventing bacterial enteritis of grass carp and preparation method thereof

Info

Publication number: CN113599432B
Application number: CN202111071446.XA
Authority: CN
Inventors: 齐琼; 魏树阁; 吕慧源; 丁曦媛; 石凯亓; 王志明; 李明; 江厚生; 王秀敏
Original assignee: Beijing Centre Biology Co ltd; Edison Beijing Biotechnology Co ltd
Current assignee: Beijing Centre Biology Co ltd; Edison Beijing Biotechnology Co ltd
Priority date: 2021-09-13
Filing date: 2021-09-13
Publication date: 2023-04-25
Anticipated expiration: 2041-09-13
Also published as: CN113599432A

Abstract

A Chinese herbal medicine composition for treating enteritis of grass carp caused by bacteria and application effect are disclosed. The Chinese herbal medicine formula comprises: eucommia ulmoides, dogwood, astragalus membranaceus, pulsatilla chinensis, phellodendron, rheum officinale, coptis chinensis, 15-35 parts of eucommia ulmoides, 10-20 parts of dogwood, 20-40 parts of astragalus membranaceus, 10-30 parts of pulsatilla chinensis, 10-30 parts of phellodendron, 10-20 parts of rheum officinale and 15-25 parts of coptis chinensis. The compound Chinese herbal medicine preparation provided by the invention can effectively improve enteritis of grass carp caused by aeromonas hydrophila, increase survival rate of grass carp, improve body surface and in vivo inflammation conditions, reduce inflammatory factors HIF-1 alpha and NF- κB of intestinal tract, and provide reference for enteritis caused by other similar pathogenic bacteria and other fishes when the addition amount of the compound Chinese herbal medicine preparation is 2%o. The compound Chinese herbal medicine preparation reasonably combines traditional Chinese medicine raw materials, has the functions of clearing heat, eliminating dampness, astringing, relieving diarrhea, inhibiting pathogenic microorganisms, improving intestinal health, has no toxic or side effect, obvious curative effect, low cost, convenient use, simple production process and wide market prospect, and provides a Chinese herbal medicine formula product for treating fish bacterial enteritis for aquaculture industry.

Description

Traditional Chinese medicine for preventing bacterial enteritis of grass carp and preparation method thereof

Technical Field

The invention belongs to the technical field of animals, and particularly relates to a Chinese herbal medicine formula for preventing bacterial enteritis of grass carp and application thereof.

Background

Grass carp is one of four traditional freshwater aquaculture fishes in China, is popular with wide consumers and fishermen, and is widely popularized and cultured. In recent years, the demand of the market for aquatic products rises year by year, the demand of grass carp also shows the trend of increasing year by year, the price is stable and medium, and the cost of green feed for cultivating grass carp is low, and the benefit is high. However, due to the lag of people's cultivation concept and the lag of aquatic disease research, and the poor disease resistance of grass carp, various bacterial diseases such as bacterial gill rot disease, saprolegniasis, red skin disease, enteritis and the like become main factors which seriously influence the healthy cultivation and restrict the improvement of the yield and economic benefit.

In production practice, bacterial diseases are one of the main diseases in the grass carp breeding process, especially bacterial enteritis of grass carp. In most of the culture areas in China, bacterial enteritis generally occurs in 4-9 months, and is extremely harmful to grass carp. Bacterial enteritis fish is characterized by red and swelling anus, protruding outwards when serious, redness of fin, redness of abdomen, inflammation of intestinal canal, purplish red erosion, poor elasticity of intestinal canal, easy tearing of intestinal canal, bloody pus caused by mixing of abdominal distention, blood and ulcerated mucous membrane bacteria, and the intestinal canal is filled with the fish. Some intestinal tracts of the diseased fish only show cream yellow mucus, and other viscera such as intestinal membranes, livers, kidneys and the like of the diseased fish are also engorged with blood and inflamed, which is the most easily dead fish in enteritis, and most of the feces are purulent and white feces and drags feces.

According to the prior art, CN201811109235 discloses a preparation for treating grass carp five diseases, which comprises the following raw materials: radix et rhizoma Rhei, cortex Phellodendri, radix Scutellariae, radix astragali, and rhizoma Coptidis; the preparation method comprises the following steps: pulverizing the above materials, decocting in water for 2 times, filtering, cooling, mixing filtrates, and mixing with water. The mixture has effects of clearing heat and detoxicating, promoting blood circulation for detumescence, resisting bacteria, astringing, purgation, removing blood stasis, increasing blood platelet, and promoting blood coagulation, and can be used for treating grass carp gill rot, enteritis, hemorrhagic disease, white head disease, white mouth disease, red swelling and pain (abstract). CN200610014333 discloses a Chinese medicinal composition for fowl, which comprises radix Pulsatillae, coptidis rhizoma, cortex Phellodendri, radix et rhizoma Rhei and radix Sophorae Flavescentis. The composition has antibacterial, organism defense enhancing, phagocyte phagocytosis enhancing, lymphocyte transformation promoting effects, is used for treating enteritis of fowl (example 1). In addition, CN201010206634 also discloses medicines such as rhubarb, phellodendron bark, etc. However, it has been found through analysis that the treatment of bacterial diseases in grass carp (or other animals) by the above-mentioned patent application (in particular CN 201811109235) is limited to clearing away heat and toxic materials, antibacterial and anti-inflammatory, but neglects the regulation of its own dysfunctions. Therefore, the symptoms and root causes are treated, the diseases are easy to relapse, and the cure rate is low.

The invention combines the modern traditional Chinese medicine theory, and the preparation prepared by screening, extracting and matching the single traditional Chinese medicine components has the advantages of small side effect, high safety, and unique self-body, and is a good substitute for antibiotics. The compatibility of the Chinese herbal medicines is targeted for different fish enteritis, so that the application technology of the Chinese herbal medicines in the formula in grass carp feed is necessary to be explored and improved.

Disclosure of Invention

The invention aims to provide a Chinese herbal medicine formula for treating grass carp bacterial enteritis, which has a good inhibition effect on pathogenic bacteria such as aeromonas hydrophila, aeromonas punctata, aeromonas guinea, and the like, and can effectively prevent and treat grass carp enteritis caused by bacteria.

The invention also aims to provide an application effect study of the Chinese herbal medicine composition in grass carp culture, and provides a reference basis for practical production and application.

The technical experimental scheme of the invention is as follows: chinese herbal medicine for treating bacterial enteritis of grass carp, which comprises the following components in parts by weight: 20-35 parts of eucommia ulmoides, 15-25 parts of dogwood, 20-30 parts of astragalus membranaceus, 10-30 parts of pulsatilla chinensis, 10-30 parts of phellodendron amurense, 10-20 parts of rheum officinale and 15-25 parts of coptis chinensis.

Further, the Chinese herbal medicine comprises the following components in parts by weight: 25-30 parts of eucommia ulmoides, 15-20 parts of dogwood, 20-25 parts of astragalus membranaceus, 25-30 parts of pulsatilla chinensis, 23-25 parts of phellodendron amurense, 15-20 parts of rheum officinale and 20-25 parts of coptis chinensis.

Further, the Chinese herbal medicine comprises the following components in parts by weight: 25 parts of eucommia ulmoides, 20 parts of dogwood, 25 parts of astragalus membranaceus, 25 parts of pulsatilla chinensis, 25 parts of phellodendron amurense, 15 parts of rheum officinale and 25 parts of coptis chinensis.

Further, the Chinese herbal medicine comprises the following components in parts by weight: 26 parts of eucommia ulmoides, 19 parts of dogwood, 24 parts of astragalus membranaceus, 26 parts of pulsatilla chinensis, 25 parts of phellodendron, 16 parts of rheum officinale and 24 parts of coptis chinensis.

Further, the Chinese herbal medicine comprises the following components in parts by weight: 28 parts of eucommia ulmoides, 17 parts of dogwood, 22 parts of astragalus membranaceus, 30 parts of pulsatilla chinensis, 23 parts of phellodendron, 17 parts of rheum officinale and 23 parts of coptis chinensis.

Further, the Chinese herbal medicine comprises the following components in parts by weight: 30 parts of eucommia ulmoides, 15 parts of dogwood, 20 parts of astragalus membranaceus, 30 parts of pulsatilla chinensis, 25 parts of phellodendron, 18 parts of rheum officinale and 22 parts of coptis chinensis.

The characteristics and functions of the components of the invention are shown in table 1.

Furthermore, the preparation method of the Chinese herbal medicine formula medicament comprises the following steps:

(1) The Chinese herbal medicine raw materials are subjected to superfine grinding and sieving,

(2) Weighing eucommia bark, dogwood, astragalus root, pulsatilla root, phellodendron bark, rhubarb and coptis root according to parts by weight respectively,

(3) Mixing, soaking in water, decocting in water, cooling,

(4) And (5) filtering after standing, and concentrating the filtrate.

Further, the screen is a 100 mesh screen.

Further, the water is 5-20 times, preferably 10 times of the weight of the traditional Chinese medicine.

Further, the soaking time is 0.5 to 24 hours, preferably 1 hour.

Further, the water decoction condition is boiling for 1.5-2 hours with slow fire.

Further, the cooling condition is that the mixture is kept stand at room temperatureCooling for 2-8 hours, preferably 4 hours。

Further, the filtration adoptsGauze or filter membrane.

Further, the concentration density is1.20±0.02。

Further, the mixture can be stored in a refrigerator at 4 ℃.

Table 1 Chinese herbal medicine composition characteristics and Functions

The traditional Chinese medicines in the formula are all natural medicines, are recorded in the two parts of the pharmacopoeia of the people's republic of China, and are legal medicines. The traditional Chinese medicine of the formula is a common traditional Chinese medicine, and can be widely used without drug residues or little residues, so that the traditional Chinese medicine can not affect people and the environment.

The composition of the feed for this experiment is shown in Table 2: according to the experiment, 2 per mill of the experimental scheme, the Chinese herbal medicine disclosed by the invention is added into basic feed, so that the dosage of rice bran serving as a feed raw material is reduced, and the composition ratio of other feed raw materials is unchanged.

TABLE 2 composition of feed materials tested

Under the condition of feeding each group of feed, the compound Chinese herbal medicine provided by the invention has obvious effect on bacterial enteritis of grass carp, treats both principal and secondary aspect of disease, has rapid effect and has no recurrence.

The preparation process of the compound Chinese herbal medicine is simple, the raw materials are low in cost, and the compound Chinese herbal medicine is suitable for large-scale industrial production popularization and application.

Drawings

FIG. 1 grass carp disease Activity index scoring

FIG. 2 HIF-1 alpha detection of intestinal health index

FIG. 3 detection of intestinal health index NF- κB

Detailed Description

The following description of the specific embodiments further illustrates and describes the technical solution of the present invention.

Example 1

(1) Respectively weighing Eucommiae cortex, corni fructus, radix astragali, radix Pulsatillae, cortex Phellodendri, radix et rhizoma Rhei, and Coptidis rhizoma whisker, and sieving with 100 mesh sieve.

(2) The crushed components are fully mixed according to 25 parts of eucommia bark, 20 parts of dogwood, 25 parts of astragalus, 25 parts of pulsatilla chinensis, 25 parts of phellodendron, 15 parts of rheum officinale and 25 parts of coptis chinensis.

(3) Mixing the weighed medicines, adding water 10 times of the weight of the medicines, and soaking for 1 hour, wherein the extraction conditions are as follows: boiling with slow fire for 1.5-2 hours; and standing and cooling for 4 hours at room temperature.

(4) And (5) concentrating the supernatant after standing until the density is 1.20+/-0.02, thus obtaining the product.

(5) The using method comprises the following steps: 2g of the product is added into 1kg of feed raw material according to the addition amount of 2 per mill, and the experimental feed is obtained by granulating after uniform mixing, wherein the diameter of the feed granule is about 5mm. Young fish 540 were selected, weighing around 50g, and randomly grouped into 6 groups of 3 replicates each, 30 fish replicates each. After 1 week of temporary culture, normal feed is fed. After the experiment is started, feeding is carried out according to the configuration feed of each group of experiments, the feeding amount is 3% -4% of the weight of the grass carp, the feeding amount is adjusted according to the feeding condition, after 1 week, the anus swab method is adopted to attack toxin, the toxin-attacking strain is aeromonas hydrophila, the death rate of each group is recorded 24 hours after the toxin attack, the dissecting observation score is sampled, the grass carp remained after the sampling is normally fed, the feeding amount is 1% -2% of the weight of the grass carp, the number of dead grass carp in each cylinder is observed and counted after the experiment is carried out for 1 week, and the death rate, the infection rate and the health amount of each group are recorded on the 1 st day. 5 fish were taken from each of the surviving fish on days 1, 3, and 7, respectively, and subjected to body surface observation, section observation, scoring, and disease activity index scoring (disease activity index, DAI) was calculated, with the scoring criteria shown in table 3. The sewage is discharged every day in the experimental period, 1 water is changed every 2 days, and the water change amount is about 1/3. The water temperature is 25-30 ℃, the ammonia nitrogen concentration is less than 2mg/L, the nitrite concentration is less than 0.01mg/L, the dissolved oxygen content is more than 5.0mg/L, the pH is 7.5-8.0, and the feeding test is 14d.

TABLE 3 Fish disease Activity index scoring criteria

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Example 2

The eucommia ulmoides, the dogwood, the astragalus membranaceus, the pulsatilla chinensis and the phellodendron amurense are respectively weighed, sieved by a 100-mesh sieve, and the crushed components are fully mixed according to 26 parts of eucommia ulmoides, 19 parts of dogwood, 24 parts of astragalus membranaceus, 26 parts of pulsatilla chinensis, 25 parts of phellodendron amurense, 16 parts of rheum officinale and 24 parts of coptis chinensis.

The preparation and use methods are the same as in example 1.

Example 3

The eucommia ulmoides, the dogwood, the astragalus membranaceus, the pulsatilla chinensis and the phellodendron amurense are respectively weighed, sieved by a 100-mesh sieve, and the crushed components are fully mixed according to 28 parts of eucommia ulmoides, 17 parts of dogwood, 22 parts of astragalus membranaceus, 30 parts of pulsatilla chinensis, 23 parts of phellodendron amurense, 17 parts of rheum officinale and 23 parts of coptis chinensis.

The preparation and use methods are the same as in example 1.

Example 4

Respectively weighing eucommia ulmoides, dogwood, astragalus mongholicus, pulsatilla chinensis and phellodendron amurense, sieving with a 100-mesh sieve, and mixing the crushed components according to eucommia ulmoides: 30 parts of dogwood: 15 parts of astragalus root: 20 parts of Chinese pulsatilla root: 30 parts of amur corktree bark 25 parts, rhubarb 18 parts and coptis root 22 parts are fully mixed.

The preparation and use methods are the same as in example 1.

Example 5

In contrast, 25 parts of astragalus, 25 parts of phellodendron, 15 parts of rheum officinale and 25 parts of coptis chinensis are respectively weighed.

The preparation and use methods are the same as in example 1.

Example 6

In contrast, 25 parts of pulsatilla chinensis, 25 parts of phellodendron, 15 parts of rheum officinale and 25 parts of coptis chinensis are respectively weighed.

The preparation and use methods are the same as in example 1.

Example 7

In contrast, 25 parts of eucommia ulmoides, 20 parts of dogwood, 25 parts of phellodendron, 15 parts of rheum officinale and 25 parts of coptis chinensis are respectively weighed.

The preparation and use methods are the same as in example 1.

Example 8

In contrast, no drug was added to the feed.

The preparation and use methods are the same as in example 1.

Example 9

In contrast, no drug was added to the feed.

The preparation method and the application method are the same as in example 1, but the aeromonas hydrophila is not used for detoxification, and the physiological saline is equal to the bacterial liquid.

The results are shown in Table 4. The result shows that the mortality rate of the negative control group of the example 8 is 63.33 and the infection rate is 36.67 when the Chinese herbal medicine formula is formed by adding different proportions in the grass carp raising process; example 9 the blank group had no death, the infection rate was 30%, and the infection was likely due to trauma to anus during injection of physiological saline or intestinal damage caused by a large amount of physiological saline entering the intestinal tract, but had little effect on overall health of fish. The Chinese herbal medicine composition of the invention has remarkable effects on preventing and treating bacterial enteritis in examples 1-4, and has remarkable effects on preventing and treating bacterial enteritis in groups compared with mortality and infection rate in examples 8 (negative) < example 6< example 5< example 7< example 3< example 2< example 1< example 4< example 9 (control), wherein the mortality rate of example 4 is 10%, the infection rate is 65.33%, the mortality rate is reduced by 53.33% compared with the negative control group, the infection rate is reduced by 18.34%, and the Chinese herbal medicine composition of the invention has good effects on treating and preventing bacterial enteritis in grass carp in groups compared with examples 5-7 in the control group.

TABLE 4 prevention and treatment effects of the Chinese herbal medicine formula of the invention on bacterial enteritis of grass carp

Figure 1 shows grass carp disease activity index scores. The results in FIG. 1 show that the disease activity indexes are scored at different periods after the challenge, and are sampled at 1, 3 and 7 days, and at 1d, the disease activity indexes of all groups have no significant difference, the disease activity indexes of examples 1-8 are all >11, and the disease activity index of example 9 (blank) is 9.2; at 3d, the disease activity index was reduced for examples 1-4 relative to examples 5-7, but there was no significant difference between the groups of examples 1-7, with example 4 being significantly reduced compared to example 8; at 7d, examples 1-3 were significantly less than example 4, and significantly less than example 8 (blank), wherein example 4 was significantly less than example 9, and examples 5-7 were significantly less than example 8. The Chinese herbal medicine formula disclosed by the invention has better prevention and treatment effects on bacterial enteritis of grass carp, wherein the Chinese herbal medicine formula in the embodiment 4 has the most obvious effect.

As can be seen from fig. 2, the results show that each of the chinese herbal medicine additive groups with intestinal related inflammatory factor HIF-1 a was significantly lower than the group of example 8 (negative), and the HIF-1 a inflammatory factor level of the group of example 4 was relatively lower than that of the other chinese herbal medicine additive groups, and significantly reduced relative to the group of example 3, and exhibited significantly reduced compared to the groups of examples 5 to 7 and example 8 (negative); as can be seen from fig. 3, the results show that the various additive groups of herbal medicines had no significant difference in the enteric inflammatory factor HIF-1 κ of examples 1-4, but significantly lower than examples 5-7, and each group was significantly lower than the group of example 8 (negative), wherein the inflammatory factor HIF-1 κ of example 4 was the lowest in expression level, and no significant difference from the group of example 9 (blank).

Fig. 3 also shows a similar trend as fig. 2.

In conclusion, the Chinese herbal medicine formula has a good control effect on enteritis caused by bacteria, and can play a role in treating both symptoms and root causes.

The above description is only a preferred implementation combination of the present invention, and the application scope of the present invention is not limited thereby, and under the principle of the present invention, the formulation may be modified in various ways, and the modifications are all within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine for preventing bacterial enteritis of grass carp is characterized in that: the traditional Chinese medicine is prepared from the following raw materials in parts by weight: 25-30 parts of eucommia bark, 15-20 parts of dogwood, 20-25 parts of astragalus, 25-30 parts of pulsatilla root, 23-25 parts of amur corktree bark, 15-20 parts of rhubarb, 20-25 parts of coptis root,

the preparation method of the traditional Chinese medicine comprises the following steps:

(3) Mixing, soaking in water, decocting in water, cooling,

(4) And (5) filtering after standing, and concentrating the filtrate.

2. The traditional Chinese medicine of claim 1, wherein: the raw material medicines comprise the following components in parts by weight: 25 parts of eucommia ulmoides, 20 parts of dogwood, 25 parts of astragalus membranaceus, 25 parts of pulsatilla chinensis, 25 parts of phellodendron amurense, 15 parts of rheum officinale and 25 parts of coptis chinensis.

3. The traditional Chinese medicine of claim 1, wherein: the raw material medicines comprise the following components in parts by weight: 26 parts of eucommia ulmoides, 19 parts of dogwood, 24 parts of astragalus membranaceus, 26 parts of pulsatilla chinensis, 25 parts of phellodendron, 16 parts of rheum officinale and 24 parts of coptis chinensis.

4. The traditional Chinese medicine of claim 1, wherein: the raw material medicines comprise the following components in parts by weight: 28 parts of eucommia ulmoides, 17 parts of dogwood, 22 parts of astragalus membranaceus, 30 parts of pulsatilla chinensis, 23 parts of phellodendron, 17 parts of rheum officinale and 23 parts of coptis chinensis.

5. The traditional Chinese medicine of claim 1, wherein: the raw material medicines comprise the following components in parts by weight: 30 parts of eucommia ulmoides, 15 parts of dogwood, 20 parts of astragalus membranaceus, 30 parts of pulsatilla chinensis, 25 parts of phellodendron, 18 parts of rheum officinale and 22 parts of coptis chinensis.

6. The method for preparing the traditional Chinese medicine according to any one of claims 1 to 5, which is characterized in that: the preparation method comprises the following steps:

(3) Mixing, soaking in water, decocting in water, cooling,

(4) And (5) filtering after standing, and concentrating the filtrate.

7. The method of manufacturing according to claim 6, wherein: the water in the step (3) is 5-20 times of the weight of the traditional Chinese medicine, the soaking time is 0.5-24 hours, and the water decocting condition is that the water is boiled for 1.5-2 hours with slow fire.