A traditional Chinese medicine composition for treating avian colibacillosis and a preparation method thereof

Abstract

The application provides a traditional Chinese medicine composition for treating avian colibacillosis and a preparation method thereof, and belongs to the technical field of traditional Chinese medicine preparations.The preparation method is as follows: 12-15 parts of Huangqin, 9-11 parts of Huanglian, 3.5-4.5 parts of Jinyinhua, 5-7 parts of Chuanxinlian and 5-7 parts of Pugongying are mixed, crushed, 8-10 times of the mass of an acidic ethanol solution is added, extracted, filtered, and a composite liquid one is obtained; 11-13 parts of Gegen, 10-12 parts of Baotouweng, 4-5 parts of Cebaiye, 6-7 parts of Wuhezi and 7-9 parts of Zixingujia are mixed, crushed, 8-10 times of the mass of an ethanol aqueous solution is added, extracted, filtered, and a composite liquid two is obtained; the composite liquid one and the composite liquid two are respectively vacuum rotary evaporated to a relative density of 1.09-1.13, mixed, and spray dried, and the traditional Chinese medicine composition is obtained.The traditional Chinese medicine composition prepared by the application has a fast effect speed and a remarkable curative effect for treating avian colibacillosis of chicks.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of traditional Chinese medicine preparation, and particularly relates to a traditional Chinese medicine composition for treating avian colibacillosis and a preparation method thereof. BACKGROUND

[0002] Avian colibacillosis is a bacterial infectious disease caused by avian pathogenic Escherichia coli infection, and all age groups of poultry are susceptible. The morbidity rate of young poultry (such as chickens, ducks and geese) and 2-6 week-old meat poultry is high. The transmission routes include vertical transmission (contamination of eggs), respiratory tract / digestive tract contact transmission, and environmental media (feces, bedding, air). The typical symptoms are depression, anorexia, abdominal distension, and yellow-greenish diarrhea. The morbidity and mortality rates are high, and the direct and indirect economic losses are huge, which seriously affects the development of the poultry industry.

[0003] At present, the main treatment for avian colibacillosis is mainly antibiotic treatment; the commonly used antibiotics are florfenicol, enrofloxacin, norfloxacin, etc. However, on the one hand, pathogenic Escherichia coli has various serotypes and is prone to mutation, and the long-term abuse of antibiotics leads to the accelerated production of drug-resistant strains, and the sensitivity of commonly used drugs such as florfenicol and enrofloxacin decreases year by year. On the other hand, the residues of antibiotics in poultry meat and eggs may enter the human body through the food chain, trigger allergic reactions, or induce drug resistance of human pathogenic bacteria, and the international standards for antibiotic residues in poultry products are becoming more stringent, and the problem of exceeding the standard directly affects the export trade.

[0004] The problems of antibiotic resistance and residues force the industry to seek alternative solutions. Traditional Chinese medicine compositions can reduce the risk of drug resistance through the synergistic effect of multiple components, and have ecological safety and immune regulation functions, and have become an important research direction for the prevention and control of poultry diseases. SUMMARY

[0005] To solve the problems in the background art, the present application provides a traditional Chinese medicine composition for treating avian colibacillosis and a preparation method thereof, which ensures that the prepared traditional Chinese medicine composition has a fast effect speed and a significant treatment effect for treating avian colibacillosis in young chickens.

[0006] To achieve the above-mentioned purpose, in a first aspect, the present application provides a preparation method of a traditional Chinese medicine composition for treating avian colibacillosis, comprising the following steps:

[0007] S1, according to the weight fraction, 12-15 parts of Huangqin, 9-11 parts of Huanglian, 3.5-4.5 parts of Jinyinhua, 5-7 parts of Chuanxinlian and 5-7 parts of Pugongying are mixed, crushed to 40-60 meshes, 8-10 times of acid ethanol solution by mass is added, ultrasonic extraction is carried out first, then microwave assisted extraction is carried out, filtration is carried out, and a composite liquid one is obtained.

[0008] The direct antibacterial (acting on bacteria) effects of each traditional Chinese medicine component are as follows:

[0009] Baicalin and Baicalein are contained in Huangqin. Baicalin can enhance the antibacterial effect by destroying the integrity of bacterial cell membrane. Baicalin can interfere with the process of bacterial DNA synthesis, indirectly affecting the function of DNA gyrase. Baicalein can reduce bacterial drug resistance through multi-target action, and can enhance the inhibitory effect on drug-resistant E. coli when combined with Berberine (Berberine).

[0010] Berberine is contained in Huanglian. Berberine can change the membrane permeability by binding to bacterial membrane phospholipids, leading to the leakage of contents. Berberine can also antagonize the activity of bacterial aldehyde synthase and inhibit the expression of quorum sensing and virulence factors, thereby weakening the metabolic adaptability of drug-resistant bacteria.

[0011] Chlorogenic acid is contained in Jinyinhua. Chlorogenic acid has antibacterial activity and can enhance the inhibitory effect on avian E. coli when combined with baicalin and Huanglian alkaloids.

[0012] Andrographolide and Neoadrographolide are contained in Chuanxinlian. Andrographolide can interfere with the bacterial quorum sensing system, reduce bacterial communication and polysaccharide matrix secretion, thereby destroying the structural stability of the biofilm. Andrographolide can also inhibit the expression of drug resistance genes and reduce the efflux of drugs by bacteria. Neoadrographolide can enhance the antibacterial activity of andrographolide by interfering with the arrangement of bacterial membrane phospholipid layer and destroying the three-dimensional structure of the biofilm.

[0013] Taraxinic acid is contained in Pugongying. Taraxinic acid can reduce bacterial colonization in host cells by inhibiting the expression of bacterial adhesion factors such as pilin. Taraxinic acid can also down-regulate the multidrug resistance genes of E. coli (such as acrAB-tolC), restoring the drug sensitivity of drug-resistant bacteria.

[0014] S2, according to the weight ratio, 11-13 parts of Gegen, 10-12 parts of Baishouweng, 4-5 parts of Cibaishan, 6-7 parts of Wuji, and 7-9 parts of Zixingjiao are mixed, crushed to 20-40 mesh, and then 8-10 times the mass of ethanol aqueous solution is added, soaked, heated and refluxed, filtered, and a compound liquid two is obtained.

[0015] Among them, the auxiliary antibacterial effects (acting on itself) of each traditional Chinese medicine component are as follows:

[0016] Puerarin is contained in Gegen. Puerarin can reduce the release of pro-inflammatory factors (such as IL-6 and TNF-α) by inhibiting the NF-kB pathway, thereby reducing intestinal inflammatory response.

[0017] Saponins, triterpenoid saponins (A3, B, B4), and carrot glycosides are active components contained in Baishouweng, which have anti-inflammatory effects.

[0018] Platycladi Cacumen contains quercitrin and tannin; it can shorten the clotting time and protect the integrity of intestinal mucosal barrier.

[0019] Galla Chinensis is rich in tannic acid; it can quickly coagulate proteins to form a protective film, and is suitable for local hemostasis and ulcer repair.

[0020] Echinacea purpurea contains polysaccharides, alkyl amides, and chicoric acid, etc., which can activate macrophages and T lymphocytes through multiple targets, and significantly enhance the non-specific immune response of chicks.

[0021] S3, vacuum rotary evaporation of the composite liquid one obtained in S1 and the composite liquid two obtained in S2 to a relative density of 1.09-1.13 (hot measurement at about 60℃) to obtain extract one and extract two.

[0022] S4, mixing the extract one and the extract two obtained in S3 to obtain a mixed extract, and spray drying, to obtain the traditional Chinese medicine composition for treating avian colibacillosis.

[0023] Further, in S1, the preparation method of the acidic ethanol solution is: adding dilute hydrochloric acid to an ethanol aqueous solution with a volume fraction of 68-72%, and adjusting the pH to 4-4.5 to obtain the acidic ethanol solution; the mass fraction of the dilute hydrochloric acid is 0.1-0.3%.

[0024] Further, in S1, the specific operation of ultrasonic extraction is: ultrasonic frequency of 40-50KHz, temperature of 60±2℃, and time of 50-60min.

[0025] Further, in S1, the specific operation of microwave assistance is: microwave power of 380-410W, temperature of 68±2℃, and time of 4-6min.

[0026] Further, in S2, the volume fraction of the ethanol aqueous solution is 50-55%.

[0027] Further, in S2, the soaking time is 5-8h, and the specific operation of heating and refluxing is: heating to 85-90℃, and refluxing for 2-2.5h.

[0028] Further, in S3, the specific operation of vacuum rotary evaporation is: rotary evaporation at 60±2℃ under a vacuum degree of ≤-0.08MPa.

[0029] Further, in S4, after adding 0.6-0.9% of silicon dioxide powder by mass in the mixed extract, spray drying is performed.

[0030] Further, the parameters of spray drying are: inlet air temperature of 110-120℃, and outlet air temperature of 65-75℃.

[0031] In a second aspect, the present application provides a traditional Chinese medicine composition for treating avian colibacillosis, which is prepared by the above method.

[0032] The present application has the following beneficial effects:

[0033] 1. In the present application, the direct antibacterial components (Scutellaria, Coptis, honeysuckle, and Andrographis) and the auxiliary antibacterial components (Pueraria, Anemone, Platycladus, and Eupatorium) are extracted separately, which can effectively avoid the loss of activity caused by cross-reaction and retain the optimal efficacy of each component through precise process; then the direct antibacterial components and the auxiliary antibacterial components are mixed for pharmaceutical use, which ensures that the final traditional Chinese medicine composition can better combine the advantages of the direct antibacterial components (fast sterilization) and the auxiliary antibacterial components (immune synergy).

[0034] 2. In the extraction process of the direct antibacterial components (Scutellaria, Coptis, honeysuckle, and Andrographis), acid ethanol solution is used; on the one hand, the acid ethanol solution can better meet the acid dissolution requirement of scutellarein in Scutellaria and the salification reaction of alkaloids in Coptis, and better meet the polarity and liposolubility requirements of flavonoid components in other medicinal materials, reducing the degradation of flavonoid components, thereby facilitating the extraction of (direct antibacterial) active ingredients;

[0035] On the other hand, the tannic acid contained in Coptis and the scutellarein contained in Scutellaria form macromolecular complexes through hydrogen bonds, hydrophobic interactions, and charge neutralization in acid ethanol solution (if it is in a non-acid environment, it is difficult to form a complex due to charge repulsion), and the molecular weight exceeds the solvent solubility, which ultimately leads to precipitation and a decrease in dissolution rate, thereby adversely affecting the extraction of (direct antibacterial) active ingredients;

[0036] The green acid contained in honeysuckle can compete with scutellarein to bind tannic acid, preferentially forming a soluble green acid-tannic acid complex, effectively reducing the loss of scutellarein, thereby effectively weakening the adverse effects; at the same time, green acid itself also has antibacterial activity, which can enhance the inhibitory effect on avian colibacillosis when combined with scutellarein and alkaloids in Coptis; this offsetting and synergistic effect can improve the sterilization speed (speed of effect) and treatment effect of the final traditional Chinese medicine composition. BRIEF DESCRIPTION OF DRAWINGS

[0037] Figure 1 In the test example of the present application, the mortality trend chart of the chicks in each group (blank control group, infection control group, example 1-3, and comparative example 1-4) after 24 hours of infection;

[0038] Figure 2 In the test example of the present application, the mortality trend chart of the chicks in each group (blank control group, infection control group, example 1-3, and comparative example 1-4) after 24 hours of infection;

[0039] Figure 3 Figure 1 is a trend chart of the cure rate of each group (Example 1-Example 3 groups and Comparative Example 1-Comparative Example 4 groups) of the test example of the present application after 7 days of infection of the chicks.

[0040] Figure 4 Figure 2 is a trend chart of the number of cures of each group (Example 1-Example 3 groups and Comparative Example 1-Comparative Example 4 groups) of the test example of the present application after 7 days of infection of the chicks. DETAILED DESCRIPTION

[0041] The technical solutions in the embodiments of the present application will be described clearly and completely below. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0042] The raw materials of the embodiments and comparative examples of the present application are all ordinary commercial products, except for special indications. Example 1

[0043] The present embodiment provides a preparation method of a traditional Chinese medicine composition for treating avian colibacillosis, comprising the following steps:

[0044] S1, according to the weight fraction, 14 parts of Huangqi, 10 parts of Huanglian, 4 parts of Jinyinhua, 6 parts of Chuanxinlian and 6 parts of Pugongying are mixed, crushed to 40-60 meshes, and a mixture one is obtained. A 9-fold mass of an acidic ethanol solution is added to the mixture one. The specific preparation method of the acidic ethanol solution is as follows: a dilute hydrochloric acid is added to an ethanol aqueous solution with a volume fraction of 70%, the mass fraction of the dilute hydrochloric acid is 0.2%, and the pH is adjusted to about 4.2, and then the acidic ethanol solution is obtained. The dilute hydrochloric acid can quickly adjust the pH to about 3.5, and has good compatibility with ethanol, without forming a precipitate or decomposition products; the hydrochloride has strong saltation effect, which is beneficial to the dissolution and stability of alkaloids (such as berberine in Huanglian); and it has less volatile residue, which does not affect the subsequent concentration / drying process. Then, it is stirred at 180 r / min for 5 min to mix evenly.

[0045] First, ultrasonic extraction is performed, and the specific operation is as follows: the ultrasonic frequency is 45 KHz, the temperature is heated to about 60℃, and the time is 55 min. Then, microwave-assisted extraction is performed, and the specific operation is as follows: the microwave power is 390 W, the temperature is heated to about 68℃, and the time is 5 min. Then, double-layer 300-mesh nylon filter cloth (wetted with pure water before use and treated with 121℃ high-pressure sterilization for 20 min) is used for filtration, and a filtrate is obtained, which is a compound solution one.

[0046] S2, by weight fraction, 12 parts of Pueraria, 11 parts of Pulsatillae Radix, 4.5 parts of Biota orientalis, 6.5 parts of Rhus and 8 parts of Echinacea purpurea are mixed, and are pulverized to 20-40 mesh to obtain a mixture two. To the mixture two, 9 times of the mass of an ethanol aqueous solution is added, and the volume fraction of the ethanol aqueous solution is 52%. Then, 180 r / min stirring is performed for 5 min to mix.

[0047] After being soaked for 7 h, heating is performed to about 88℃, and reflux is performed for 2.2 h. Then, double-layer 300 mesh nylon filter cloth (wetted with pure water before use and treated with 121℃ high-pressure sterilization for 20 min) is used for filtration to obtain a filtrate, which is a compound liquid two.

[0048] S3, the compound liquid one obtained in S1 and the compound liquid two obtained in S2 are vacuum rotary evaporated respectively, and the specific operation is as follows: rotary evaporation is performed at about 60℃ under the condition of vacuum degree-0.08 MPa. Both are vacuum rotary evaporated to a relative density of about 1.11 (hot measurement at about 60℃), and two kinds of extract are obtained respectively.

[0049] S4, the two kinds of extract obtained in S3 are mixed to obtain a mixed extract. After adding 0.8% of the mass of silicon dioxide powder to the mixed extract and mixing, spray drying is performed, and the specific operation is as follows: a centrifugal atomizing disc is used, the rotating speed is 18000 rpm, the particle size D50=80-100 μm, the inlet air temperature is about 115℃, the outlet air temperature is about 70℃, the feeding speed is 2.2 L / h (controlled by a peristaltic pump), the drying air flow is 35 m 3 / h (maintaining negative pressure-50 Pa), and a traditional Chinese medicine composition for treating avian colibacillosis is obtained.

[0050] The silicon dioxide powder is pharmaceutical grade silicon dioxide (AEROSIL ® 200 Pharma), which is purchased from Yinkang Special Chemical (Shanghai) Co., Ltd. Example 2

[0051] The difference between this example and example 1 is that this example provides a preparation method of a traditional Chinese medicine composition for treating avian colibacillosis, which comprises the following steps:

[0052] S1, by weight fraction, Scutellaria 12 parts, 9 parts of Rhizoma Coptidis, honeysuckle 3.5 parts, 5 parts of Andrographis paniculata and 5 parts of dandelion mixed, crushed to 40-60 mesh, to get the mixture one. To the mixture one is added 8 times its mass of acidic ethanol solution, the acidic ethanol solution is prepared by adding 0.1% dilute hydrochloric acid to the volume fraction of 68% ethanol aqueous solution to adjust the pH to about 4.2. Stirring at 180 r / min for 5 min to mix. First ultrasonic extraction, the specific operation is: ultrasonic frequency is 40KHz, the temperature is about 60℃, the time is 60 min. Then microwave assisted, the specific operation is: microwave power is 380W, the temperature is about 68℃, the time is 6 min. Then double 300 mesh filter cloth filtration, get the filtrate, for the composite liquid one.

[0053] S2, by weight fraction, 11 parts of Radix Puerariae, 10 parts of Radix Pulsatillae, 4 parts of Platycladus orientalis, 6 parts of gallnut and 7 parts of Eupatorium fortunei mixed, crushed to 20-40 mesh, to get the mixture two. To the mixture two is added 8 times its mass of ethanol aqueous solution, the volume fraction of ethanol aqueous solution is 50%. Stirring at 180 r / min for 5 min to mix. After soaking for 5 h, heat to 90℃, reflux for 2 h. Then double 300 mesh filter cloth filtration, get the filtrate, for the composite liquid two.

[0054] S3, the composite liquid one obtained in S1 and the composite liquid two obtained in S2 are vacuum rotary evaporated respectively, the specific operation is: under the condition of vacuum degree-0.08MPa, 60℃ rotary evaporation. Vacuum rotary evaporation to the relative density of 1.11 or so (60℃ hot measurement), respectively, to get the extract one and the extract two.

[0055] S4, the extract one and the extract two obtained in S3 are mixed to get the mixed extract. After adding 0.8% of silicon dioxide powder to the mixed extract, spray drying is carried out, to get the traditional Chinese medicine composition for treating avian colibacillosis. Example 3

[0056] The difference between this embodiment and example 1 is that this embodiment provides a preparation method of a traditional Chinese medicine composition for treating avian colibacillosis, comprising the following steps:

[0057] S1, by weight parts, Huangqi 13 parts, Baishu 12 parts, Caibaiye 5 parts, Wupeizi 7 parts and Zixiangju 9 parts are mixed, and are crushed to 20-40 meshes to obtain mixture two. 10 times the mass of ethanol solution is added to the mixture two, and the volume fraction of the ethanol solution is 55%. Stirring at 180 r / min for 5 min to mix. After soaking for 8 h, heat to 85℃, and reflux for 2.5 h. Then double 300 mesh filter cloth is filtered to obtain the filtrate, which is a composite liquid two.

[0058] S2, by weight parts, Huangqi 13 parts, Baishu 12 parts, Caibaiye 5 parts, Wupeizi 7 parts and Zixiangju 9 parts are mixed, and are crushed to 20-40 meshes to obtain mixture two. 10 times the mass of ethanol solution is added to the mixture two, and the volume fraction of the ethanol solution is 55%. Stirring at 180 r / min for 5 min to mix. After soaking for 8 h, heat to 85℃, and reflux for 2.5 h. Then double 300 mesh filter cloth is filtered to obtain the filtrate, which is a composite liquid two.

[0059] S3, the composite liquid one obtained in S1 and the composite liquid two obtained in S2 are vacuum rotary evaporated respectively, and the specific operation is as follows: under the condition of vacuum degree-0.08 MPa, rotary evaporation at about 60℃. The relative density is about 1.11 (hot measurement at about 60℃), and the extract one and the extract two are obtained respectively.

[0060] S4, the extract one and the extract two obtained in S3 are mixed to obtain a mixed extract. After adding 0.8% of silicon dioxide powder to the mixed extract, spray drying is carried out, and a traditional Chinese medicine composition for treating avian colibacillosis is obtained.

[0061] Comparative Example 1:

[0062] The difference between this comparative example and Example 1 is only that in S1, the acidic ethanol solution is replaced by a 70% ethanol aqueous solution by volume fraction; and the Honeysuckle is deleted.

[0063] Comparative Example 2:

[0064] The difference between this comparative example and Example 1 is only that in S1, the Honeysuckle is deleted.

[0065] Comparative Example 3:

[0066] The difference between this comparative example and Example 1 is only that in S1, the acidic ethanol solution is replaced by a 70% ethanol aqueous solution by volume fraction.

[0067] Comparative Example 4:

[0068] The difference between the present comparative example and Example 1 is that the gallnut in S2 is replaced by extraction in S1.

[0069] Specifically, the present comparative example provides a preparation method of a traditional Chinese medicine composition for treating avian colibacillosis, comprising the following steps:

[0070] S1, by weight fraction, Scutellaria 14 parts, Coptis 10 parts, honeysuckle 4 parts, gallnut 6.5 parts, Andrographis 6 parts and dandelion 6 parts are mixed, crushed to 40-60 mesh, to obtain mixture one. Add 9 times the mass of the acid ethanol solution to the mixture one. Then mix evenly by stirring at 180 r / min for 5 min. First ultrasonic extraction, then microwave assisted, then double 300 mesh nylon filter cloth filtration, to obtain the filtrate, which is the composite liquid one.

[0071] S2, by weight fraction, Pueraria 12 parts, Anemone 11 parts, Platycladus 4.5 parts and Echinacea 8 parts are mixed, crushed to 20-40 mesh, to obtain mixture two. Add 9 times the mass of the ethanol aqueous solution to the mixture two, and the volume fraction of the ethanol aqueous solution is 52%. Then mix evenly by stirring at 180 r / min for 5 min. First soak, then heat reflux, then double 300 mesh nylon filter cloth filtration, to obtain the filtrate, which is the composite liquid two.

[0072] S3, the composite liquid one obtained in S1 and the composite liquid two obtained in S2 are vacuum rotary evaporated respectively, and both are vacuum rotary evaporated to a relative density of about 1.11 (about 60°C hot measurement), to obtain extract one and extract two respectively.

[0073] S4, the extract one and the extract two obtained in S3 are mixed to obtain a mixed extract. After adding 0.8% of silicon dioxide powder to the mixed extract and mixing, spray drying is performed, to obtain the traditional Chinese medicine composition for treating avian colibacillosis.

[0074] Test example:

[0075] Test object: the traditional Chinese medicine composition for treating avian colibacillosis prepared in Example 1-Example 3 and Comparative Example 1-Comparative Example 4.

[0076] Test method: test grouping: 270 chicks (4-day-old Hyline laying hens) were randomly divided into 9 groups, which were blank control group, infection control group, Example 1-Example 3 group and Comparative Example 1-Comparative Example 4 group, 30 in each group.

[0077] Test procedure: in turn, infection, treatment and data statistics, as follows: ① Infection: except the blank control group, the rest 8 groups of chicks were intraperitoneally injected with avian pathogenic E. coli strain 1.0×10 8 CFU / one (injection volume is 0.2 mL), and the blank control group was injected with the same amount of normal saline.

[0078] 2. Treatment: After 6h of infection, the groups of Example 1-Example 3 and the groups of Comparative Example 1-Comparative Example 4 were respectively orally administered with the corresponding traditional Chinese medicine composition 0.03g / bird (dissolved in normal saline, the administration volume was 0.25mL), the blank control group and the infection control group were orally administered with normal saline 0.25mL / bird, and each group was continuously administered for 5 days (once a day).

[0079] 3. Data statistics of morbidity after 6h of infection, mortality after 24h of infection, and cure rate after 7d of infection: the symptoms of the chicks included listlessness, drooping wings, loose and messy feathers, neck retraction, walking after being driven, immobility after being driven, lying on the ground, water-like feces, feces contamination of the anus and surrounding feathers, etc., which were determined as morbidity. The disappearance of clinical symptoms and no obvious pathological changes in dissection were determined as cure.

[0080] After 6h of infection, the morbidity after 6h of infection was determined by the ratio of the number of chicks with morbidity in each group to the total number of chicks in the group; that is, the morbidity after 6h of infection = the number of chicks with morbidity after 6h of infection / the total number of chicks in the group x 100%.

[0081] After 24h of infection, the mortality after 24h of infection was determined by the ratio of the number of dead chicks in each group to the total number of chicks in the group; that is, the mortality after 24h of infection (%) = the number of dead chicks after 24h of infection / the total number of chicks in the group x 100%.

[0082] After 7d of infection, the cure rate after 7d of infection was determined by the ratio of the number of cured chicks in each group to the total number of chicks in the group; that is, the cure rate after 7d of infection (%) = the number of cured chicks after 7d of infection / the total number of chicks in the group x 100%.

[0083] Test results: see Table 1.

[0084] Table 1. Test data of test examples:

[0085]

[0086] Result analysis: analyzing the blank control, the infection control, and Example 1-Example 3, and combining the data in Table 1 and Figures 1-4 It can be seen that the traditional Chinese medicine composition prepared by the present application (Example 1-Example 3) can reduce the mortality of chicks after 24h of infection to below 13.3%, and the cure rate after 7d of infection is as high as 86.7%, and all the chicks that do not die in the early stage (within 24h) of infection are cured. It shows that the traditional Chinese medicine composition prepared by the present application (Example 1-Example 3) has a fast effect and a significant effect for treating colibacillosis in chicks.

[0087] Analyzing Example 1 and Comparative Example 1-Comparative Example 4, and combining the data in Table 1 and Figures 1-4Compared with the comparative example 1 and the comparative example 2, it can be seen that, compared with the comparative example 1 (not containing honeysuckle), the extraction solution of the direct antibacterial component in step S1 is replaced by the acidic ethanol solution of the present application (i.e. the pH of the ethanol aqueous solution is adjusted to about 4.5 by using dilute hydrochloric acid) in the comparative example 2; the mortality of the chicks after 24 hours of infection is increased from 30% (comparative example 1) to 36.7% (comparative example 2), and the cure rate of the chicks after 7 days of infection is decreased from 60% (comparative example 1) to 46.7% (comparative example 2). It is shown that, when the extraction solution of the direct antibacterial component (not containing honeysuckle) in step S1 is replaced by the acidic ethanol solution of the present application, the prepared traditional Chinese medicine composition for treating avian colibacillosis of chicks has a decreased effective speed and final curative effect.

[0088] This is because, in the extraction process of the direct antibacterial component (scutellaria, coptis, and andrographis), the acidic ethanol solution is used; on the one hand (favorable effect), the acidic ethanol solution can better take into account the acidic dissolution requirement of baicalin in scutellaria and the salification reaction of alkaloids in coptis, and better take into account the polarity and fat solubility requirement of other medicinal materials flavonoids, and reduce the degradation of flavonoids, thereby being beneficial to the extraction of effective components (direct antibacterial). On the other hand (unfavorable effect), the tannic acid contained in coptis and the baicalin contained in scutellaria form macromolecular complexes through hydrogen bonds, hydrophobic interaction and charge neutralization in the acidic ethanol solution (if it is in a non-acidic environment, it is difficult to form a complex due to charge repulsion), and the molecular weight exceeds the solvent solubility, which eventually leads to precipitation and a decrease in dissolution rate, thereby being unfavorable to the extraction of effective components (direct antibacterial). The degree of unfavorable effect is greater than that of favorable effect, and finally causes negative effects on the performance (effective speed and curative effect) of the prepared traditional Chinese medicine composition.

[0089] Compared with the comparative example 1 and the comparative example 3, it can be seen that, compared with the comparative example 1, the honeysuckle is added to the direct antibacterial component in the comparative example 3, and the mortality of the chicks after 24 hours of infection is decreased from 30% (comparative example 1) to 26.7% (comparative example 3), and the cure rate of the chicks after 7 days of infection is increased from 60% (comparative example 1) to 66.7% (comparative example 3). It is shown that, when the extraction solution is the ethanol aqueous solution, the addition of honeysuckle to the direct antibacterial component can improve the effective speed and final curative effect of the prepared traditional Chinese medicine composition for treating avian colibacillosis of chicks.

[0090] In combination with the example 1, it can be seen that, the replacement of the extraction solution of the direct antibacterial component by the acidic ethanol solution of the present application and the addition of honeysuckle to the direct antibacterial component can produce a synergistic effect, and synergistically improve the effective speed and final curative effect of the prepared traditional Chinese medicine composition for treating avian colibacillosis of chicks.

[0091] This is because the greenish acid contained in honeysuckle can compete with baicalin to bind to tannic acid in Huanglian, and preferentially form a soluble greenish acid-tannic acid complex, avoiding the formation of a macromolecular complex of tannic acid and baicalin in acidic ethanol solution, effectively reducing the loss of baicalin, retaining the beneficial effects of acidic ethanol solution while effectively weakening the adverse effects; at the same time, greenish acid itself also has antibacterial activity, and can enhance the inhibitory effect on avian E. coli when combined with baicalin and alkaloids in Huanglian, thereby synergistically improving the speed of effect and the final curative effect of the prepared traditional Chinese medicine composition for treating avian E. coli disease in chicks.

[0092] By comparing Example 1 and Comparative Example 4, it can be seen that, compared with Example 1, in Comparative Example 4, the gallnut in the auxiliary antibacterial component in step S2 is replaced by being extracted together with the direct antibacterial component in step S1, and the mortality rate of chicks after 24h of infection increases from 13.3% (Example 1) to 20% (Comparative Example 4), and the cure rate of chicks after 7d of infection decreases from 86.7% (Example 1) to 73.3% (Comparative Example 4). This shows that replacing the gallnut in the auxiliary antibacterial component with the direct antibacterial component will result in a decrease in both the speed of effect and the final curative effect of the prepared traditional Chinese medicine composition for treating avian E. coli disease in chicks. The inventors speculate that this is because the high tannic acid content of gallnut causes the negative effects of the formation of a macromolecular complex of tannic acid and baicalin in acidic ethanol solution to be difficult to be effectively weakened by honeysuckle (greenish acid).

[0093] In addition, it should be noted that each specific technical feature described in the above specific embodiments can be combined in any appropriate manner without contradiction, and in order to avoid unnecessary repetition, the present application will not further describe various possible combinations.

[0094] In addition, various different embodiments of the present application can also be combined in any appropriate manner, as long as they do not deviate from the inventive concept of the present application, and they should also be considered as disclosed by the present application.

Claims

1. A method for preparing a traditional Chinese medicine composition for treating avian colibacillosis, characterized in that, It comprises the following steps: S1, by weight parts, Scutellaria 12-15 parts, 9-11 parts of Huanglian, 3.5-4.5 parts of honeysuckle, 5-7 parts of Andrographis, and 5-7 parts of dandelion are mixed, crushed, 8-10 times the mass of acid ethanol solution is added, ultrasonic extraction is carried out first, then microwave assisted, filtered, and composite liquid one is obtained; S2, by weight parts, 11-13 parts of Radix Puerariae, 10-12 parts of Anemone, 4-5 parts of Platycladus orientalis, 6-7 parts of gallnut and 7-9 parts of Echinacea purpurea are mixed, crushed, 8-10 times the mass of ethanol aqueous solution is added, soaked, heated to reflux, filtered, and composite liquid two is obtained; S3, the composite liquid one obtained in S1 and the composite liquid two obtained in S2 are vacuum rotary evaporated to relative density of 1.09-1.13, and extract one and extract two are obtained; S4, the extract one and the extract two obtained in S3 are mixed to obtain mixed extract, and spray drying is carried out, thereby obtaining the traditional Chinese medicine composition for treating avian colibacillosis; In S1, the preparation method of the acid ethanol solution is as follows: adding dilute hydrochloric acid to ethanol aqueous solution with a volume fraction of 68-72%, and adjusting the pH to 4-4.5, thereby obtaining the acid ethanol solution; the mass fraction of the dilute hydrochloric acid is 0.1-0.3%; In S2, the volume fraction of the ethanol aqueous solution is 50-55%; In S2, the soaking time is 5-8h, and the specific operation of heating to reflux is as follows: heating to 85-90℃, and refluxing for 2-2.5h.

2. The method of claim 1, wherein the Chinese medicine composition for treating avian colibacillosis is prepared by mixing the following ingredients: In S1, the specific operation of ultrasonic extraction is as follows: ultrasonic frequency is 40-50KHz, temperature is 60±2℃, and time is 50-60min. ​ 3. The method for preparing the traditional Chinese medicine composition for treating avian colibacillosis according to claim 1, characterized in that, In S1, the specific operation of microwave assisted is as follows: microwave power is 380-410W, temperature is 68±2℃, and time is 4-6min.

4. The method for preparing the traditional Chinese medicine composition for treating avian colibacillosis according to claim 1, characterized in that, In S3, the specific operation of vacuum rotary evaporation is as follows: rotary evaporation is carried out at 60±2℃ under the condition of vacuum degree≤-0.08MPa.

5. The method for preparing the traditional Chinese medicine composition for treating avian colibacillosis according to claim 1, characterized in that, In S4, after adding 0.6-0.9% of silicon dioxide powder to the mixed extract, spray drying is carried out.

6. The method for preparing the traditional Chinese medicine composition for treating avian colibacillosis according to claim 1 or 5, characterized in that, The parameters of spray drying are as follows: inlet air temperature of spray drying is 110-120℃, and outlet air temperature is 65-75℃.

7. A traditional Chinese medicine composition for treating avian colibacillosis, characterized in that, The preparation method is prepared by using any one of claims 1-6. The preparation method is prepared by using any one of claims 1-6.

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