CN109453257B

CN109453257B - Traditional Chinese medicine composition and soft extract for preventing and treating poultry newcastle disease

Info

Publication number: CN109453257B
Application number: CN201811558691.1A
Authority: CN
Inventors: 汤法银; 樊克锋; 王坤丽; 王长林; 汤垚鑫
Original assignee: Henan University of Animal Husbandry and Economy
Current assignee: Henan University of Animal Husbandry and Economy
Priority date: 2018-12-19
Filing date: 2018-12-19
Publication date: 2021-07-20
Anticipated expiration: 2038-12-19
Also published as: CN109453257A

Abstract

The invention discloses a traditional Chinese medicine composition (pestilence and detoxication) and a decoction paste for preventing and treating poultry newcastle disease, which are prepared from the following raw material medicines in parts by weight: 8-15 parts of isatis root, 1-5 parts of honeysuckle, 3-7 parts of scutellaria baicalensis, 3-7 parts of gardenia, 3-7 parts of radix rehmanniae, 3-7 parts of pokeberry root, 5-15 parts of astragalus mongholicus and 3-7 parts of liquorice. The decoction ointment which has stable property, large drug-loading rate, easy industrial production, full ingredient retention and capability of being administrated by drinking water in groups is selected, and the pestilent toxin clearing decoction ointment disclosed by the invention is subjected to heat understanding, anti-inflammatory and immunity enhancement and curative effect tests on artificial infection of newcastle disease of chickens.

Description

Traditional Chinese medicine composition and soft extract for preventing and treating poultry newcastle disease

Technical Field

The invention belongs to the field of traditional Chinese veterinary medicines, and particularly relates to a traditional Chinese medicine composition (pestilence and toxin removing) and a decoction paste for treating poultry Newcastle disease.

Background

Newcastle disease is a highly contagious, lethal disease caused by Newcastle Disease Virus (NDV) that mainly attacks chickens, turkeys, wild birds, and ornamental birds, and is classified as a class a infectious disease that presents the greatest harm to animals by the international veterinary community. The new castle disease is popular in China for many years, is one of the most important virulent infectious diseases harming the poultry industry in China, brings huge loss to the poultry industry, and is a hot disease highly concerned by the veterinary community. The current prevention and treatment means for the Newcastle disease comprise vaccine prevention, western medicine treatment, traditional Chinese medicine treatment, yolk antibody treatment, hyperimmune serum treatment and the like. The traditional Chinese medicine treatment is a means for preventing and treating animal diseases under the guidance of the theory of Chinese veterinarian, has the advantages of naturality, pluripotency, low toxicity and side effects, difficult generation of harmful residues and drug resistance and the like compared with western medicines, and is widely applied to animal clinic.

At present, the main formulation of the traditional Chinese veterinary medicine preparation for treating the Newcastle disease in the market is big, thick and black powder, the using amount is large, the medicine is mainly taken by mixing materials, however, sick livestock and poultry usually like drinking water but do not take food, and even if the livestock and poultry take food, the bioavailability is extremely low because the active ingredients exist in plant cells; the powder is purified and prepared into a mixture (oral liquid), and the oral liquid has the problems of precipitation and the like due to instability to light and heat; in order to ensure the clarity, alcohol precipitation is mostly adopted in the preparation process, and some substances with pharmacodynamic activity, such as polysaccharide, protein, amino acid and the like, are removed by the alcohol precipitation, so that the efficacy is obviously reduced; other formulations such as tablets, injections, etc. are not suitable for mass administration.

In view of the above, the development of new veterinary drug preparations suitable for group administration, stable in property, high in bioavailability and convenient to use is urgent, and in addition, the existing veterinary drug preparations for treating newcastle diseases mostly have the problems of incomplete formula, unclear treatment mechanism and the like.

Disclosure of Invention

In order to solve the technical problems, the invention provides a traditional Chinese medicine composition and a soft extract for preventing and treating the poultry Newcastle disease, which are suitable for group administration, have high bioavailability, convenient use and obvious and durable curative effect.

The inventor carries out the debate of warm diseases, qi, ying and blood and the debate of eight principles aiming at the characteristics of the viral newcastle disease of livestock and poultry, clarifies the external manifestation, the symptom characteristics and the pathogenesis of the disease, and determines the total pathological mechanism: the disease is epidemic heat toxin, dual blaze qi and blood, the syndrome type in the early stage is excess heat of qi and blood, and the syndrome type in the later stage is deficiency of both qi and blood, which is the starting point, and combines a large amount of research experiences to establish the therapeutic principle as follows: the invention relates to a Chinese medicinal composition for treating poultry newcastle disease, which is prepared from the following raw materials in part by weight:

the traditional Chinese medicine composition for preventing and treating the poultry Newcastle disease is designed and comprises the following raw material medicines in parts by weight:

8-15 parts of isatis root, 1-5 parts of honeysuckle, 3-7 parts of scutellaria baicalensis, 3-7 parts of gardenia, 3-7 parts of radix rehmanniae, 3-7 parts of pokeberry root, 5-15 parts of astragalus mongholicus and 3-7 parts of liquorice.

Preferably, the traditional Chinese medicine composition for preventing and treating the poultry newcastle disease comprises the following raw material medicines in parts by weight:

8-12 parts of isatis root, 1-3 parts of honeysuckle, 3-5 parts of scutellaria baicalensis, 3-5 parts of gardenia, 3-5 parts of radix rehmanniae, 3-5 parts of pokeberry root, 5-10 parts of astragalus mongholicus and 3-5 parts of liquorice.

12-15 parts of isatis root, 3-5 parts of honeysuckle, 5-7 parts of scutellaria baicalensis, 5-7 parts of gardenia, 5-7 parts of radix rehmanniae, 5-7 parts of pokeberry root, 10-15 parts of astragalus mongholicus and 5-7 parts of liquorice.

12 parts of isatis root, 3 parts of honeysuckle, 5 parts of scutellaria baicalensis, 5 parts of gardenia, 5 parts of radix rehmanniae, 5 parts of pokeberry root, 10 parts of astragalus membranaceus and 5 parts of liquorice.

The preparation method of the traditional Chinese medicine composition for preventing and treating the poultry newcastle disease, which is prepared into the soft extract, comprises the following steps:

(1) weighing the raw material medicines according to the proportion;

(2) uniformly mixing the raw materials, adding water of which the weight is 6-10 times of the total weight of the raw materials, and soaking for 1-2 hours; then decocting the raw materials in water for 1-4 times, combining decoctions, and concentrating;

(3) and (3) melting borneol, adding the melted borneol into the step (2), and concentrating.

Preferably, in the preparation method of the soft extract for preventing and treating the newcastle disease of the poultry, in the step (2), water which is 8 times of the total weight of the medicinal materials is added and soaked for 1.5 hours; then decocting with water for 3 times.

Preferably, in the step (3), 3g of the decocted liquid is prepared for every 1mL of the decocted liquid.

In addition, the invention researches the selection of the dosage form of the traditional Chinese medicine composition (pestilence and detoxication) for preventing and treating the newcastle disease of the poultry.

The existing veterinary drug formulations in clinical application mainly comprise: powder, tablets, drinking water agents (oral liquid, soluble powder and decoction paste), injection and the like, and intensive culture needs to be mainly used for group control in a prevention and treatment mode, so the dosage form should be mainly selected by mixing materials or drinking water. Because the epidemic febrile disease livestock such as sick chicken show the characteristic of preferring drinking and anorexia after suffering from the disease, and the active ingredients of most medicines in the prescription of the invention are water-soluble, the preparation formulation is positioned in a drinking water agent. The traditional Chinese veterinary medicine drinking agent mainly comprises oral liquid, but the oral liquid is often unstable to light and heat, and is easy to precipitate, and the like, and meanwhile, the oral liquid needs to be subjected to alcohol precipitation for removing impurities, namely, components such as protein, polysaccharide and the like are removed, but main active components of tonifying medicines in the formula are amino acids and polysaccharides, such as astragalus polysaccharide, rehmannia polysaccharide and the like, which need to be reserved, so that the influence of the traditional Chinese medicine composition for preventing and treating the poultry newcastle disease on the antibody titer of the poultry newcastle disease before and after alcohol precipitation is researched.

Alcohol precipitation and impurity removal are carried out, and then the preparation of the toxin-warming and toxin-clearing decocted extract is carried out: taking a proper amount of the above Wen DU QING decocted extract, adding 60% ethanol by volume ratio, mixing uniformly, standing overnight, and filtering to obtain supernatant; adding 60% ethanol into the residue, filtering to obtain supernatant, mixing the two supernatants, and recovering ethanol until the volume of the supernatant is equal to that of the original Wenxuqing decoction.

150 test chickens of 60 days old are randomly divided into 5 groups, one group is a control group, the other 4 groups are test groups, each chicken of the test groups I, II and III respectively takes water drinking and drug feeding of 0.5g, 1.0g and 1.5g of the warm-toxin decoction cream, each chicken of the test groups IV respectively takes water drinking and drug feeding, alcohol precipitation and impurity removal of 1.0mL of the warm-toxin decoction cream, and on the basis of the original immunity, 0.5mL of ND-Lasota series vaccine is injected. Blood was collected at the corresponding time, and the titer of anti-newcastle disease antibody was measured by Hemagglutination Inhibition (HI) assay, and ND antibody was measured after 1 week, 2 weeks, and 3 weeks of immunization in test groups I, II, III, and IV, respectively, and the specific results are shown in Table 1.

TABLE 1 comparison of the effects of pestilence virus-clearing on the antibody titer of newcastle disease in chickens

Note: indicates significant difference (P <0.05) and indicates very significant difference (P < 0.01).

SAS statistical software is adopted for statistical analysis, t-test analysis is carried out on experimental data, and it can be seen from measured values that the antibody level reaches the peak value 2 weeks after immunization, and each test group has obvious difference compared with a control group after 2 weeks, which shows that the specific antibody level can be obviously improved before and after the mild toxicity cleaning decoction is subjected to alcohol precipitation; the difference between the test groups II and III and the control group is very obvious (P is less than 0.01), which shows that the level of the specific antibody can be obviously improved by the Wen Du Qing decocted extract, and the level of the specific antibody is increased along with the increase of the dosage of the Wen Du Qing decocted extract; compared with the test group IV, the test groups II and III have obvious difference (P is less than 0.05), which shows that the effect of improving the level of the specific immune antibody by the warm toxin removing after alcohol precipitation and impurity removal is reduced.

The research result proves that: the temperature-clearing decoction paste has the effect of remarkably improving the valence level of the newcastle disease antibody before and after alcohol precipitation, and the immune enhancement effect without alcohol precipitation and impurity removal in the preparation process is more prominent. Therefore, the soft extract which has stable property, large drug-loading rate, easy industrialized production and full ingredient retention and can be administrated by drinking water in groups is selected as a final preparation form and is named as the warm toxin clearing soft extract. And the preparation formulation further conforms to the overall syndrome differentiation treatment principle of multiple components, multiple ways, multiple targets and multiple functions of Chinese veterinary medicines.

The pharmacological and medicinal effects and the compatibility mechanism of the traditional Chinese medicine are as follows:

based on the pathogenesis characteristics of poultry Newcastle disease ' warm epidemic heat toxin, dual blaze qi and blood ', the invention has the advantages of clearing heat and detoxicating, purging fire and cooling blood, and strengthening body resistance and eliminating evil ', and the invention has the following components:

in the formula, the radix isatidis is used as a monarch drug for directly clearing excess heat in qi system and having the function of protecting body fluid, and the honeysuckle is added for lightly clearing and ventilating, dispelling heat and externally reaching the effect of clearing heat and ventilating heat toxin in exterior and interior;

the auxiliary materials are scutellaria baicalensis, gardenia and the like, the excess fire of the triple energizer is released, and the monarch and ministerial drugs are combined, so that the effect of clearing heat in the qi system is achieved, and the triple energizer is released;

the rehmannia root and the pokeberry root are used together as an adjuvant, and are specially used for cooling blood and removing toxicity, nourishing yin and removing blood stasis to clear heat of blood, and the cape jasmine and the pokeberry root are used together to clear heart and induce diuresis, and conduct heat downward;

radix astragali, radix rehmanniae and liquorice are used for replenishing qi and blood, so that the healthy qi is sufficient and the evil cannot dry, so as to improve the immunity of the organism and enhance the antibacterial and antiviral effects of the organism, and the liquorice is matched for clearing and moistening the throat and treating sore throat;

in addition, the compound preservative such as borneol is favorable for relieving exterior syndrome, refreshing mind, inducing resuscitation and relieving sore throat due to the cool property, and has good natural preservative effect.

The medicines are compatible and combined, the formula mainly takes the heat clearing, detoxifying, dampness eliminating and heat clearing away from the triple warmer of the white tiger decoction and the coptis chinensis detoxification decoction, and the traditional Chinese medicine has a strong inhibiting effect on the Newcastle disease virus; taking the horn and rehmannia decoction for soothing liver and calming endogenous wind, cooling blood and promoting fluid production as auxiliary materials, nourishing yin and removing blood stasis to clear heat and clear qi and blood; the astragalus and the liquorice are used for assisting in invigorating spleen and tonifying qi and harmonizing drug properties, and can obviously improve the immune function of the organism, and the medicines are combined, so that the traditional Chinese medicine composition can effectively inhibit virus proliferation, can also adjust the yin-yang balance of the organism, enhances the immune capability, and achieves the aim of treating both symptoms and root causes.

Compared with the prior art, the invention has the beneficial technical effects that:

1. the pestilence and toxin clearing decoction paste has good antipyretic and anti-inflammatory effects and immunity enhancing effect, and can be used for preventing and treating newcastle disease infectious diseases of poultry.

2. The pestilence toxin clearing decoction ointment is suitable for group administration, stable in property, large in drug loading rate, easy for industrial production, full-component retention, uniform and stable.

3. The pestilent toxin clearing decoction ointment has the advantages of ingenious formula, reasonable compatibility of medicines, convenience in use and lasting and obvious curative effect.

Detailed Description

The invention is further illustrated by the following specific examples in order to enable those skilled in the art to better understand the invention, but without thereby restricting it.

Example 1

A traditional Chinese medicine composition (pestilence and toxin removing soft extract) for preventing and treating poultry Newcastle disease is prepared from the following raw materials in parts by weight:

12 parts of isatis root, 3 parts of honeysuckle, 5 parts of scutellaria baicalensis, 5 parts of gardenia, 5 parts of radix rehmanniae, 5 parts of pokeberry root, 10 parts of astragalus mongholicus and 5 parts of liquorice, and the specific preparation method is shown in example 7.

Example 2

8 parts of isatis root, 1 part of honeysuckle, 3 parts of scutellaria baicalensis, 3 parts of gardenia, 3 parts of radix rehmanniae, 3 parts of pokeberry root, 5 parts of astragalus mongholicus and 3 parts of liquorice, and the specific preparation method is shown in example 7.

Example 3

15 parts of isatis root, 5 parts of honeysuckle, 7 parts of scutellaria baicalensis, 7 parts of gardenia, 7 parts of radix rehmanniae, 7 parts of pokeberry root, 15 parts of astragalus mongholicus and 7 parts of liquorice, and the specific preparation method is shown in example 7.

Example 4

15 parts of isatis root, 3 parts of honeysuckle, 7 parts of scutellaria baicalensis, 5 parts of gardenia, 7 parts of radix rehmanniae, 5 parts of pokeberry root, 15 parts of astragalus membranaceus and 5 parts of liquorice, and the specific preparation method is shown in example 7.

Example 5

isatis root 10 parts, honeysuckle flower 5 parts, baikal skullcap root 5 parts, cape jasmine 7 parts, dried rehmannia root 5 parts, pokeberry root 7 parts, astragalus root 10 parts, licorice root 7 parts, the concrete preparation method is referred to example 7.

Example 6

isatis root 10 parts, honeysuckle flower 1 part, baikal skullcap root 5 parts, cape jasmine 3 parts, dried rehmannia root 5 parts, pokeberry root 3 parts, astragalus root 10 parts, licorice root 3 parts, the concrete preparation method is referred to example 7.

Example 7

The traditional Chinese medicine composition for preventing and treating poultry newcastle disease in the embodiment 1-6 is prepared into a decoction (pestilence toxin removing decoction), and comprises the following steps:

(1) preparing the raw material medicines according to the proportion;

(2) uniformly mixing the raw materials, adding water of which the weight is 6-10 times of the total weight of the raw materials, and soaking for 1-2 hours; then decocting the raw materials in water for 2-4 times, combining decoctions, and concentrating;

(3) and (3) melting borneol, adding the borneol into the step (2), concentrating at low temperature under negative pressure, and preparing 3g of decocted liquid per 1 mL.

A great number of experiments prove that the optimal process of the traditional Chinese medicine composition (pestilence and toxicity removing decoction paste) for preventing and treating the poultry Newcastle disease is to add 8 times of water for 1.5 hours and extract the mixture by decocting for 3 times.

The pharmacological effects of the pestilent toxin clearing decoction paste of the invention are further explained below.

1. Test for antipyretic action

1.1 Effect of Wen Du Qing decocted extract on rat fever caused by endotoxin of Escherichia coli

15 rats were randomly divided into 3 groups, namely a blank control group, a treatment group and a positive control group, and each group contains 5 rats. Performing intragastric administration on the blank control group according to 10mL of distilled water per kilogram of body weight; the treatment group is administrated by stomach irrigation according to 3mL of the warm toxin clearing decoction cream per kilogram of body weight; the positive control group was given distilled water as the first group. Rectal thermometry was used before dosing, and 2d normal body temperature was measured for each group, and the average value was taken to discard rats with body temperatures outside the normal range. Each group is administrated for 3 days, each group is injected with 75 mug/kg of escherichia coli endotoxin in the abdominal cavity 1 hour after the administration on the third day, a positive control group is injected with 2mL/kg of antongding immediately after the escherichia coli endotoxin is injected with the same dose, the body temperature of a rat is measured 1, 3, 5, 7 and 24 hours after the endotoxin is given, the data analysis adopts SAS statistical analysis software to carry out statistical analysis results to carry out the inter-group difference t test, and the test results are shown in table 1.1.

The results in table 1.1 show that the Wen Du Qing decocted extract has obvious inhibition effect on the body temperature rise of rats caused by escherichia coli endotoxin at 3h and 24h after administration; the temperature-toxin-clearing decoction paste can slowly reduce the temperature rise of rats caused by the endotoxin of escherichia coli.

TABLE 1.1 Effect of Wen Du Qing decocted extract on the increase of body temperature of rats caused by endotoxin in E.coli

Note: + represents a significant difference (p ≦ 0.05) from the control group, and ++ represents a very significant difference (p <0.01) from the control group

1.2 Effect of Wen Du Qing decoction on the body temperature of rats induced by peptone

15 white rats were randomly divided into three groups of 5 rats each with the same administration route, administration dose and time of 1.1, 10% peptone was injected into each group 1h after the last administration, the body temperature of the rats was measured at different times after peptone administration, data analysis was performed by statistical analysis using SAS statistical software, and the results of t-test for differences between groups were shown in Table 1.2.

1.2 Effect of Wen Du Qing decoction on peptone-induced increase in rat body temperature

Note: (+ means significant difference from the control group (p ≦ 0.05); + + means significant difference from the control group (p <0.01)

The results in Table 1.2 show that the Wen Du Qing decoction has obvious inhibition effect on the body temperature rise of rats caused by peptone at 3 hours and 5 hours after the administration for 3 days; the result shows that the warm toxin clearing decoction has slow inhibition effect on the body temperature rise of rats caused by peptone.

2. Test for anti-inflammatory action

2.1 the effect of the pestilent toxin-clearing decoction paste on the granuloma of a rat cotton ball

Healthy wistar rats with body weights of 140-l to 60g and males are randomly divided into 6 groups. Blank control group: distilled water 10mL/kg d; cortisone oxide group: 30 mg (6 mL)/kg. d; shuanghuanglian oral liquid group: 6 mL/kg. d: high dose group of warm toxin decoction: 3 g/kg. d; the traditional Chinese medicine composition comprises the following components in dosage group: 2 g/kg. d; wen du qing decocted extract low dose group: 1 g/kg. d. Animals of each group were anesthetized with ketamine hydrochloride (0.1 g/kg, jp), hair cut at left and right groves, sterilized with iodine, deiodinated with 75% alcohol, each cut a small opening, subcutaneous tissue was expanded with vascular clamps, a sterile cotton ball (20 mg, penicillin 160U, streptomycin 0.1mg) was embedded in each side, and the skin was sutured. From the day of operation, hydrocortisone groups were administered by subcutaneous injection, the other groups were administered by intragastric administration, 1 time per day for 7 days continuously, cervical dislocation was sacrificed on day 8, cotton balls were taken out, adipose tissues were removed, dried in an oven at 60 ℃, weighed, and the weight of original cotton balls was subtracted to obtain the granuloma weight, and the granuloma index (mg/100g body weight) was calculated. Data analysis SAS statistical software is adopted to carry out statistical analysis results to carry out inter-group difference t test.

2.1 Effect of Wen Du Qing decocted extract on rat Cotton ball granuloma hyperplasia

Group of	n	Dosage form	Granuloma index (mg/100)
				Blank control group	12		28.46±11.70
Hydrocortisone group	11	0.03g	17.46±5.52
				Shuanghuanglian oral liquid group	10	10mL	20.73±8.32
High-dose group of Wen Du Qing decocted extract	11	6g	18.81±6.02*
				Middle dose group of mild poison decoction paste	11	4	18.23±5.69*
Low-dose group of Wen Du Qing decocted extract	11	2	26.23±6.37

Note: p <0.05, p <0.01 compared to the blank control.

The results are shown in table 2.1, the Shuanghuanglian oral liquid group, the Wenxuqing decoction paste with high and medium dosage and hydrocortisone have obvious inhibition effect on the granuloma gain of the cotton balls of rats, and have significant difference compared with a blank control group (P <0.05 and P < 0.01). The low dose group has a tendency of inhibiting the cotton ball granuloma hyperplasia of the rat, but has no statistical significance (P is more than 0.05').

2.2 Effect of the pestilence toxin-clearing decoction on the Permeability of capillary vessels in abdominal cavity of mice caused by acetic acid

Healthy mice are taken, the weight of the mice is 18-20 g, and the mice are divided into 5 groups randomly. Blank control group: distilled water 10mL/kg d; shuanghuanglian oral liquid group: 10 mL/kg. d; high dose group of warm toxin decoction: 6 g/kg. d; the traditional Chinese medicine composition comprises the following components in dosage group: 4 g/kg. d; wen du qing decocted extract low dose group: 2 g/kg. d. The preparation is administered by intragastric administration, 1 time per day, 7 days continuously, 30min after the last administration, 0.5% Evans blue normal saline (0.1mL/10g) is injected into tail vein, 0.2 mL/one of 0.6% acetic acid normal saline is injected into abdominal cavity, 20min later, mice are dislocated and killed after cervical vertebra, abdominal cavity skin muscle is cut, 6mL normal saline is used for washing abdominal cavity for several times, washing liquid is sucked out by a suction tube, normal saline is added to l0mL after combination, 3000rpm is centrifuged for 15min, supernatant is taken, and absorbance (A value) is measured at 590nm wavelength of a spectrophotometer. Data analysis SAS statistical software is adopted to carry out statistical analysis results to carry out inter-group difference t test.

The results are shown in table 2.2, and the high, medium and low doses of the Wen Du Qing decoction paste and the Shuanghuanglian oral liquid have obvious inhibition effects on the increase of the permeability of the capillary vessels in the abdominal cavity of the mouse caused by acetic acid, and have obvious statistical differences compared with a blank control group.

TABLE 2.2 Effect of pestilent toxin-clearing decoction on the increase of permeability of capillary vessels in abdominal cavity of mice caused by acetic acid

Group of	n	Dosage form	A value (. times.100)
				Blank control group	12		22.63±10.62
Shuanghuanglian oral liquid group	10	10mL	8.73±3.12**
				Medicine for warming and removing toxic matterHigh dose decoction of ointment	11	6g	8.81±2.02**
Middle dose group of mild poison decoction paste	10	4	8.73±2.66**
				Low-dose group of Wen Du Qing decocted extract	11	2	8.13±2.57**

Note: p <0.01 compared to the blank control.

The above results show that: the pestilence toxin clearing decoction has good antipyretic and anti-inflammatory effects and immunity enhancing effect, and can be used for preventing and treating infectious diseases such as Newcastle disease. The cooling effect is as follows: the pestilence toxin clearing decoction has slow and lasting inhibition effect on two fever models, namely an escherichia coli endotoxin fever model and a peptone fever model; the anti-inflammatory effect is as follows: the pestilence toxin clearing decoction paste has obvious inhibition effect on rat cotton ball granuloma hyperplasia and mouse abdominal cavity capillary permeability enhancement caused by acetic acid, and has obvious difference compared with a blank control.

3. Research on influence of pestilence toxin clearing decoction paste on immune function of broiler chicken

Selecting 400 feathers of 1-day-old Avermectins broiler chicken, and providing the feathers by a breeding farm of southern river agricultural university. Test chickens were randomly divided into four groups, three groups of test groups, a low dose group, a medium dose group, a high dose group and a control group, each group had 25 feathers, and each group had four replicates. The test broiler chickens were fed according to normal standards and were fed with free water. Three warm-toxin-clearing decoction pastes with different doses are respectively added into drinking water of broilers in a test group, namely a high-dose group (1500 g/T), a medium-dose group (1000 g/T) and a low-dose group (500 g/T); the control group had water normally. Each group was vaccinated at 7d with newcastle disease vaccine (purchased from midrange stock gmbh) and at 14d for a second immunization. Selecting 5 feather test chickens from each group of 10d, 20d, 30d and 40d, aseptically taking blood, weighing, killing, weighing bursa of fabricius, and measuring immune organ index: weighing the bursa of Fabricius, spleen and thymus of the slaughtered test chickens at 1d, 10d, 20d, 30d and 40d after being dried by filter paper, calculating immune organ indexes, wherein the immune organ indexes (g/kg) = organ weight/body weight, performing statistical analysis on data by using SAS statistical software, and expressing the data as the average +/-standard error (X +/-sE).

The bursa of fabricius is a specific central organ of humoral immunity of poultry, and is a main place for the differentiation and development of B cells. After the embryo at later stage or the hatching chick is cut off the upper bursa of the cavity, the corresponding immune function is reduced or disappeared, the cells (secreting antibody) are reduced or disappeared, the specific antibody can not be generated after the antigen stimulation, and the humoral immunity is inhibited. The thymus is the central organ for avian cellular immunity and is the main site for T cell differentiation and development. If the thymus of a newborn animal is excised, the animal lacks T cells and the immune function is significantly reduced. The spleen is the largest peripheral immune organ of poultry, and the spleen of poultry is small in size and mainly involved in humoral immunity and cellular immunity. Therefore, the development of immune organs is closely related to the strength of the immune function of the body. Currently, thymus, spleen and bursal index are commonly used to reflect the immune function status of the body.

3.1 Effect of pestilence toxin-clearing decoction on bursa of Fabricius index of test chickens

The test results are shown in Table 3.1.

TABLE 3.1 Effect of pestilent toxicity-removing decoction on bursa of Fabricius index of test chickens

Group of	1d	10d	20d	30d	40d
						Control group	1.41±0.025^a	1.52±0.032^a	1.93±0.065^a	2.21±0.078^a	1.98±0.059^a
Low dose group	1.39±0.028^a	1.65±0.038^b	2.14±0.058^b	2.62±0.046^c	2.51±0.055^c
						Middle dose group	1.43±0.031^a	1.65±0.037^b	2.18±0.074^b	2.71±0.053^c	2.58±0.069^c
High dose group	1.35±0.032^a	1.68±0.040^b	2.23±0.046^c	2.69±0.087^c	2.59±0.043^c

Note: the adjacent ones with the shoulder mark letter in the same column indicate significant difference (P <0.05), the alternate ones with the letter indicate significant difference (P <0.01), and the ones with the same letter indicate insignificant difference (P > 0.05).

As can be seen from Table 3.1, the bursa of Fabricius index of the 10d tested chickens is remarkably lower than that of each test group (P <0.05), and the difference between each test group is not remarkable (P > 0.05); the bursa of Fabricius index of the chicken in the 20d test is remarkably lower than that of a low-dose group and a medium-dose group (P <0.05), is remarkably lower than that of a high-dose group (P <0.01), is not remarkably different from that of the low-dose group and the medium-dose group (P > 0.05), but is remarkably lower than that of the high-dose group (P < 0.05); the bursa of Fabricius index of chickens tested at 30d and 40d is remarkably lower than that of each experimental group (P <0.01), and the difference between each experimental group is not remarkable (P > 0.05).

3.2 Effect of Wen Du Qing decocted extract on thymus index of test chicken

The test results are shown in Table 3.2, and it can be seen that the breast gland index (g/kg) of the 10d test chicken is significantly lower than that of each test group (P <0.05), and the difference between each test group is not significant (P > 0.05); the thymus index of 20d, 30d and 40d test chickens is remarkably lower than that of each test group (P <0.01) in the control group, and the difference between the test groups is not remarkable (P > 0.05).

TABLE 3.2 comparison of the effect of the Wen Du Qing decoction on the thymus index of the tested chickens

Group of	1d	10d	20d	30d	40d
						Control group	1.33±0.042^a	1.58±0.054^a	3.95±0.106^a	3.88±0.125^a	2.42±0.096^a
Low dose group	1.36±0.047^a	1.74±0.066^b	4.60±0.132^c	4.62±0.148^c	3.65±0.125^c
						Middle dose group	1.32±0.037^a	1.76±0.049^b	4.54±0.156^c	4.72±0.109^c	3.66±0.113^c
High dose group	1.33±0.039^a	1.76±0.036^b	4.73±0.168^c	4.69±0.137^c	3.72±0.163^c

3.3 Effect of Wen Du Qing decoction on spleen index of test Chicken

The effect of the Wen DU QING decocted extract on the spleen index is shown in Table 3.3. It can be seen that the thymus index of 10d, 20d, 30d and 40d tested chickens is remarkably lower than that of each test group (P <0.01) in the control group, and the difference between the test groups is not remarkable (P > 0.05).

TABLE 3.3 comparison of the influence of the Wen Du Qing decoction on the thymus index of the tested chickens

Group of	1d	10d	20d	30d	40d
						Control group	0.44±0.018^a	0.56±0.033^a	1.34±0.049^a	1.75±0.061^a	1.94±0.088^a
Low dose group	0.47±0.016^a	0.69±0.056^c	1.81±0.095^c	2.34±0.095^c	2.41±0.102^c
						Middle dose group	0.45±0.015^a	0.77±0.025^c	1.87±0.047^c	2.40±0.063^c	2.52±0.057^c
High dose group	0.43±0.014^a	0.73±0.061^c	1.91±0.077^c	2.41±0.086^c	2.56±0.089^c

The test result shows that the immune organ index can be obviously or extremely obviously improved after the warm toxin removing decocted extract is drunk for 10 days and 20 days, and the immune organ index can be extremely obviously improved after the warm toxin removing decocted extract is drunk for 30 days and 40 days.

3.4 Effect of Wen Du Qing decoction on test Chicken lymphocyte transformation ratio

Lymphocyte transformation rate assay lymphocyte transformation rates were determined by the MTT method, canavalin (ConA) and MTT were purchased from Sigma, RP1640 medium from invit-rogen. The results of the determination of the conversion rate of the warm-toxin decoction paste to the test chicken lymphocytes are shown in Table 1.4.

As can be seen from table 3.4, the lymphocyte conversion rates of the broiler chickens tested at 10d and 20d were significantly lower in the control group than in each test group (P <0.01), significantly lower in the low dose group than in the medium dose group and the high dose group (P <0.05), and not significantly different between the medium dose group and the high dose group (P > 0.05); the lymphocyte conversion rate of the broiler chickens in the test of 30 days is remarkably lower than that of a low-dose group (P <0.05), is remarkably lower than that of a medium-dose group and a high-dose group (P <0.01), and has no remarkable difference between the medium-dose group and the high-dose group (P > 0.05); the lymphocyte conversion rate of the test broiler chickens in 40 days is remarkably lower than that of a low-dose group (P <0.05), is extremely remarkably lower than that of a medium-dose group and that of a high-dose group (P <0.01), and has remarkable difference between the test groups (P < 0.05).

The level of lymphocyte transformation is often used as a measure of the level of cellular immune function in the body. High lymphocyte transformation rate indicates that the cellular immune function of the organism is strong, and conversely, indicates that the cellular immune function of the organism is weakened. The research shows that the low-dose group can obviously or extremely obviously improve the 10d, 20d, 30d and 40d lymphocyte transformation rates, and the medium-dose group and the high-dose group can extremely and obviously improve the lymphocyte transformation rates of all ages of days.

TABLE 3.4 comparison of the Effect of Wen Du Qing decoction on the lymphocyte transformation efficiency of test chickens

Group of	1d	10d	20d	30d	40d
						Control group	19.4±1.2^a	19.4±1.2^a	28.3±3.7^a	39.2±6.1^a	43.8±4.6^a
Low dose group	19.1±1.4^a	36.1±3.4^c	45.5±3.8^c	52.6±5.5^b	59.2±4.2^b
						Middle dose group	19.4±1.2^a	49.4±2.0^d	60.9±4.2^d	71.4±3.3^c	72.1±2.5^c
High dose group	19.9±1.8^a	51.9±3.8^d	60.1±5.1^d	69.0±5.6^c	86.2±2.4^d

3.5 detection of antibody levels in serum

The antibody level in serum is determined by semi-micro hemagglutination inhibition assay (HI) with HI antibody titer in Log₂ ^XAnd (4) showing. The ND antibody titer in serum was tested by a semi-microscale hemagglutination inhibition assay, and the results are shown in Table 3.5. As can be seen from Table 1.5, the antibody titer between groups has no obvious difference (P > 0.05) when the chickens are tested for 10 days; 20. at 30 days and 40 days, the difference between the control group and the test group is very obvious (P <0.01), and the difference between the test groups is not obvious (P > 0.05).

The level of antibody production after the body has been injected with the vaccine may reflect the humoral immune function of the body. The research proves that the antibody level of the body injected with ND vaccine for a period of time can be remarkably improved after drinking warm-toxin-clearing decoction paste of each dosage group.

TABLE 3.5 Effect of Wen Du Qing decoction on the antibody titer of ND in serum

Group of	1d	10d	20d	30d	40d
						Control group	5.6±0.54^a	3.9±0.50^a	6.5±0.73^a	6.6±0.75^a	6.1±0.82^a
Low dose group	5.1±0.38^a	4.1±0.34^a	8.3±0.32^c	8.5±0.29^c	8.3±0.44^c
						Middle dose group	5.3±0.33^a	4.0±0.20^a	8.2±0.28^c	8.6±0.17^c	8.4±0.38^c
High dose group	5.8±0.35^a	3.8±0.38^a	8.2±0.34^c	8.5±0.47^c	8.5±0.25^c

The above research results show that: compared with the control group, the three indexes of immune organ index, lymphocyte transformation rate and antibody level in serum of each experimental group are obviously different (P <0.05), the medium dosage (1000 g/T) is the most appropriate addition amount, the immune organ index can be obviously or extremely obviously improved by drinking the temperature toxin clearing decoction cream for 10 days and 20 days, and the immune organ index can be extremely obviously improved by drinking the temperature toxin clearing decoction cream for 30 days and 40 days.

4. The curative effect test of the pestilence virus clearing decoction paste on Newcastle Disease Virus (NDV) artificially infected chickens:

200 test chickens of 1 day old are taken and normally bred, blood is collected once every 3 days, serum is separated, the titer (HI) of the Newcastle disease antibody in the serum is measured, weak chicks are removed when the HI is changed to 0, and the rest chickens are divided into 6 groups, namely a low-dose group, a medium-dose group, a high-dose group, a drug control group, a positive control group and a negative control group, wherein each group comprises 30 chickens. The pestilence and toxicity removing decoction cream is added into clean drinking water according to the dosage of 0.5 (low dosage), 1.0 (medium dosage) and 1.5 g/patient (high dosage) respectively for treatment group I, treatment group II and treatment group III, the administration is divided into 2 times, and the medicine is continuously taken for 5 days in a morning and evening free drinking water mode; drug control group, each administered with amantadine 0.2 mg/day, for the same time and method as the previous three groups; the positive control group only attacks poison and does not take medicine; the negative control group was not administered for toxicity. The chick is raised for 1 week and then tested, namely, the newcastle disease virus is inoculated at the age of 21 days, the NDV wild strain is diluted by 50 times, 0.1ml of nose drops are spotted on eyes per virus diluent, and the virus disease symptoms are mainly: depression, gradual decline of appetite, rough and lusterless feathers, lethargy, yellow-green loose stool discharge, crop expansion, outflow of a large amount of mucus from the oral cavity of a sick chicken, mouth opening respiration and appearance of nervous symptoms of a few sick chickens.

After attacking, the clinical symptoms and pathological changes of all groups of chickens are observed, the severity of the diseases is compared, the number of diseases, recovery and death is recorded, the time (namely the incubation period) of the symptoms appearing after attacking is observed until 15 days later, and the observation is finished.

4.1 minimum lethality assay

The minimum lethal dose measurement is carried out by using the allantoic fluid of the Newcastle disease virus rejuvenated by the chick embryos, the test result shows that the minimum lethal dose is 0.1ml per eye drop nose after being diluted by 50 times of the allantoic fluid, and the test result is shown in a table 4.1.

TABLE 4.1 comparison of the results of the efficacy tests on the newcastle disease infected chickens

Group of	Low dose group	Middle dose group	High dose group	Drug control group	Positive control group	Negative control group
							Chicken (only)	30	30	30	30	30	30
Number of sick chickens (only)	20	14	15	21	30	0
							Number of dead chicken (only)	5	2	2	6	28	0
Number of recovery chickens (only)	11	12	13	12	—	—
							Incidence (%)	66.7^a	46.7^b	50.0^b	70.0^a	100^c	—
Mortality (%)	25.0^a	14.3^b	13.3^b	28..6^c	93.3^d	—
							Recovery rate (%)	55.5^a	85.7^b	86.7^b	57.1^a	—	—

Note: in the table, the same letters in the superscript letters between the data in the same row indicate that the difference is not significant (P > 0.05), and the different letters indicate that the difference is significant (P < 0.05).

As can be seen from Table 4.1, all the chickens in the negative control group were normal and did not die, and all the chickens in the negative control group died after the infection of the negative control group for 4 days after the challenge and all the chickens in the negative control group died after 9 days. The incidence of the chickens in each treatment group is remarkably lower than that of the positive control group, the difference is very remarkable (P is less than 0.01), the lowest (46.7%) of the medium-dose group (1.0 g/chicken) is obtained, and the difference between the high-dose group and the medium-dose group is not remarkable (P is more than 0.05%) of the high-dose group, the low-dose group and the drug control group (70.0%) are obtained; compared with the cure rate and the death rate of the treatment groups, the treatment groups are very different from a positive control group (P is less than 0.01), wherein the difference between the dose group and the high dose group is not significant (P is more than 0.05), and the difference is significant (P is less than 0.05) compared with the low dose group and the drug control group, and the results show that the warm-toxicity cleaning decoction paste has obvious treatment effect on sick chickens artificially infected with the newcastle disease, and the treatment effect of the medium dose group (1.0 g/chicken) is the best.

4.2 clinical manifestations and pathological anatomy changes

All dead chickens can be seen after the dissection, mucus adheres to the oral cavity and throat, the mucous membrane of the throat is congested, and part of the chickens have bleeding; the howline cyst wall is edematous, a layer of white exudate is arranged in the howline cyst, and the howline cyst is filled with acid and odor liquid and gas; bleeding from the papilla and mucous membranes of the glandular stomach is evident, bleeding and ulceration of the muscular stomach, bleeding from the mucous membranes and serosa of the intestinal tract is evident, especially in the duodenum. Enlargement of the cecum tonsil, hemorrhage and necrosis. The low, medium and high dose groups all had lighter lesions than the positive control group.

4.3 determination of antibody levels in serum

The antibody level in serum is determined by semi-micro hemagglutination inhibition assay (HI) with HI antibody titer in Log₂ ^XAnd (4) showing. The antibody titer determination result is shown in table 4.2, the HI titer of the treatment group is significantly higher than that of the positive control group, wherein the medium dose group (2 lg 2) and the high dose group (1.7 lg 2) are the highest, which indicates that the warm toxin decoction cream has an obvious effect of inhibiting the proliferation of the newcastle disease virus in vivo.

TABLE 4.2 comparison of the results of the inhibition of the Wen Du Qing decoction on the proliferation of Newcastle disease virus

Note: p <0.05, P <0.01 in comparison with the infected group

It can be seen from tables 4.1 and 4.2 that the medium and high dose groups of the pestilent and venomous decoction cream are superior to amantadine in both experimental newcastle disease treatment effect and in vivo newcastle disease virus proliferation inhibition effect, and the difference between the medium and high dose groups is not significant. The traditional Chinese medicine formula has good prevention and treatment effects on the Newcastle disease virus infection, has good curative effects on other viruses, and can be widely popularized and applied clinically.

The results of experiments on the heat-clearing effect, the anti-inflammatory effect and the immunity enhancement effect of the pestilence toxin clearing decoction paste and the curative effect of the chicken infected with newcastle disease artificially show that the heat-clearing and toxicity-clearing decoction paste has better antipyretic and anti-inflammatory effects and immunity enhancement effects and can be used for preventing and treating infectious diseases such as newcastle disease and the like.

Although the present invention has been described in detail with reference to the embodiments, it will be understood by those skilled in the art that various changes in the specific parameters of the embodiments may be made without departing from the spirit of the present invention, and a plurality of specific embodiments are formed, which are common variations of the present invention, and will not be described in detail herein.

Claims

1. The traditional Chinese medicine composition for preventing and treating the poultry Newcastle disease is characterized by comprising the following raw material medicines in parts by weight:

2. The traditional Chinese medicine composition for preventing and treating poultry newcastle disease according to claim 1, is characterized by comprising the following raw material medicines in parts by weight:

3. The traditional Chinese medicine composition for preventing and treating poultry newcastle disease according to claim 1, is characterized by comprising the following raw material medicines in parts by weight:

4. The traditional Chinese medicine composition for preventing and treating poultry newcastle disease according to claim 1, is characterized by comprising the following raw material medicines in parts by weight:

5. A preparation method of a soft extract for preventing and treating poultry newcastle disease is characterized by comprising the following steps:

(1) weighing the raw material medicines according to the proportion of claim 1;

6. The method for preparing a soft extract for preventing and treating poultry newcastle disease according to claim 5, wherein in the step (2), water with the weight 8 times of the total weight of the medicinal materials is added for soaking for 1.5 hours; then decocting with water for 3 times.

7. The method for preparing a soft extract for preventing and treating poultry newcastle disease according to claim 5, wherein in the step (3), 3g of soft extract is prepared for every 1mL of decoction.