Acetylcysteine soluble powder and application thereof

Abstract

The invention relates to the technical field of veterinary drugs, and provides acetylcysteine soluble powder and application thereof. Research finds that acetylcysteine can improve the sensitivity of chicken mycoplasma to antibiotics, so that the acetylcysteine soluble powder is provided and comprises the following components: 10%-50% of acetylcysteine, 1%-10% of a metal complexing agent, 2%-12% of a cosolvent, 0.001%-0.01% of an antioxidant, 0.1%-2% of a flavoring agent, 0.001%-0.1% of a preservative and the balance of a filler. The acetylcysteine soluble powder not only can significantly improve the treatment effect of antibiotics on chicken mycoplasmosis, but also can reduce the occurrence of drug resistance of pathogenic bacteria.

Description

Technical Field

[0001] The present application relates to the technical field of veterinary drugs and provides an acetylcysteine ​​soluble powder and an application thereof. Background Art

[0002] Mycoplasma disease in chickens is a contact infectious disease of chickens caused by mycoplasma infection. The main pathogens include Mycoplasma Galliscepticum and Mycoplasma synoviae. Among them, Mycoplasma Galliscepticum can cause chronic respiratory diseases, leading to slow growth, decreased egg production, reduced hatching rate of breeding eggs and decreased quality of chicken carcasses. And Mycoplasma synoviae mainly causes infectious synovitis, air sac inflammation, abnormal eggshell top, increased shell breakage, and slow decline in egg production. Mycoplasma disease in chickens occurs almost all over the world and has become an important problem for chickens, causing harm to the health of chickens and causing great losses to the development of the poultry industry. The methods commonly used to control mycoplasma disease in chickens include quarantine and elimination of positive chickens for purification, antibiotic prevention and treatment, and vaccine immunization. Due to the high positive rate of breeder farms (generally 20-80%), the cost of elimination and purification is too high. In clinical practice, a combination of antibiotics and vaccine immunization is often used to prevent and treat mycoplasma disease in chickens. Commonly used antibiotics include enrofloxacin, tylosin, doxycycline and tylosin. However, with the increase in the use of antibiotics, the abuse and irrational use of antibiotics in farms have occurred from time to time, which has led to a gradual decrease in the sensitivity of livestock and poultry pathogens to antibiotics, and the antibiotic resistance has become increasingly serious. In an increasing number of farms, although mycoplasma specific drugs (such as enrofloxacin) have been used, the occurrence of chicken mycoplasma disease cannot be controlled. Therefore, how to improve the sensitivity of resistant strains to antibiotics has become an urgent issue. Summary of the invention

[0003] The purpose of the present application is to provide a new application of acetylcysteine ​​and to provide a soluble acetylcysteine ​​powder to alleviate the drug resistance problem of chicken mycoplasma in the prior art and to have a better clinical efficacy on chicken respiratory diseases caused by various other pathogens.

[0004] In order to achieve the above purpose, this application adopts the following technical solutions:

[0005] Application of acetylcysteine ​​in the preparation of drugs for improving the antibiotic sensitivity of chicken mycoplasma.

[0006] In some embodiments, the antibiotic comprises at least one of tiamulin, doxycycline, tylosin, or enrofloxacin.

[0007] In some embodiments, the content of acetylcysteine ​​is not less than 1 μg / ml, preferably (1-4) μg / ml.

[0008] In some embodiments, the Mycoplasma gallinarum includes Mycoplasma gallisepticum and Mycoplasma synoviae.

[0009] The invention discloses an acetylcysteine ​​soluble powder, which is composed of the following components by mass percentage: 10% to 50% of acetylcysteine, 1% to 10% of a metal complexing agent, 2% to 12% of a solubility aid, 0.001% to 0.01% of an antioxidant, 0.1% to 2% of a flavoring agent and 0.001% to 0.1% of a preservative, and the balance is a filler.

[0010] In some embodiments, the composition is composed of the following components, calculated by mass percentage: 15% to 30% acetylcysteine, 2% to 6% metal complexing agent, 3% to 8% co-solvent, 0.002% to 0.006% antioxidant, 0.2% to 1% flavoring agent and 0.005% to 0.05% preservative, and the balance is filler.

[0011] In some embodiments, the composition is composed of the following components by mass percentage: 20% acetylcysteine, 4% metal complexing agent, 5% co-solvent, 0.004% antioxidant, 0.5% flavoring agent and 0.01% preservative, and the balance is filler.

[0012] In some embodiments, the metal complexing agent is disodium ethylenediaminetetraacetate; and / or,

[0013] The cosolvent is at least one of anhydrous citric acid, polyethylene glycol, sodium xylene sulfonate, sodium lauryl sulfate, PEG6000, PVPK30, sodium citrate, and poloxamer 188, preferably sodium xylene sulfonate; and / or,

[0014] The antioxidant is L-ascorbic acid; and / or,

[0015] The flavoring agent is one of vanillin, cream flavor, and lemon flavor, preferably vanillin; and / or,

[0016] The preservative is sorbic acid; and / or,

[0017] The filler is one of anhydrous glucose, sucrose and anhydrous lactose, preferably anhydrous lactose.

[0018] A method for preparing acetylcysteine ​​soluble powder comprises the following steps:

[0019] Acetylcysteine, a metal complexing agent and a cosolvent are uniformly mixed to obtain an intermediate 1;

[0020] The antioxidant, the flavoring agent, the preservative and a part of the anhydrous lactose are premixed, and then the premix is ​​mixed evenly with the remaining anhydrous lactose by a doubling method to obtain an intermediate 2;

[0021] The intermediate 1 and the intermediate 2 are mixed evenly to obtain acetylcysteine ​​soluble powder.

[0022] Application of acetylcysteine ​​soluble powder in preparing medicine for auxiliary treatment of chicken mycoplasma disease.

[0023] Application of acetylcysteine ​​soluble powder in preparing medicine for auxiliary treatment of chicken respiratory diseases;

[0024] Respiratory diseases in chickens include infectious bronchitis, avian influenza or air sac disease.

[0025] Compared with the prior art, the technical effects of this application are:

[0026] Experiments have shown that acetylcysteine ​​can increase the sensitivity of chicken mycoplasma to antibiotics, and thus provide an acetylcysteine ​​soluble powder, which can not only significantly improve the therapeutic effect of antibiotics on chicken mycoplasma disease, but also reduce the occurrence of pathogen resistance, and at the same time can also reduce the viscosity of respiratory secretions, promote sputum discharge, and has a better clinical effect on chicken mycoplasma disease. In addition, the acetylcysteine ​​soluble powder can also be used as an auxiliary for chicken respiratory diseases caused by other causes, and has a better clinical effect on respiratory diseases caused by various pathogens. DETAILED DESCRIPTION

[0027] The technical solution of the present application will be described clearly and completely below in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in the field without creative work are within the scope of protection of the present application.

[0028] Acetylcysteine ​​is a thiol-containing antioxidant with expectorant and antioxidant effects. It has been used as a mucolytic for various respiratory diseases and is a traditional expectorant. It is used as a mucolytic for the treatment of various respiratory diseases and is widely used in clinical medicine for the treatment of respiratory infectious diseases. It reduces the viscosity of sputum, exerts an expectorant effect, and helps improve lung function. However, there is no prior art research on the use of acetylcysteine ​​to increase the sensitivity of Mycoplasma galli pathogens to antibiotics.

[0029] The present application proposes a new use of acetylcysteine, which can be used as a drug to increase the sensitivity of Mycoplasma gallinarum to antibiotics. Therefore, acetylcysteine ​​can be used in combination with antibiotics for treating Mycoplasma gallinarum to treat Mycoplasma gallinarum.

[0030] In some embodiments, the antibiotic can be an antibiotic commonly used to treat chicken mycoplasmosis, such as tylosin and its pharmaceutically acceptable salts (tylosin tartrate), tiamulin and its pharmaceutically acceptable salts (tiamulin fumarate), doxycycline and its pharmaceutically acceptable salts (doxycycline hydrochloride, etc.), enrofloxacin, etc.

[0031] In some embodiments, experiments show that when the content of acetylcysteine ​​is not less than 1 μg / ml, the sensitivity of Mycoplasma gallisepticum pathogen to antibiotics can be significantly improved. For example, the minimum inhibitory concentration of enrofloxacin to Mycoplasma gallisepticum is 0.25 μg / ml. When the content of acetylcysteine ​​is 1 μg / ml, the minimum inhibitory concentration can be reduced to 0.03 μg / ml. Preferably, the content of acetylcysteine ​​is (1-4) μg / ml.

[0032] In some embodiments, the Mycoplasma Galli includes Mycoplasma gallisepticum and Mycoplasma synoviae.

[0033] The present application also provides an acetylcysteine ​​soluble powder, which is composed of the following components in percentage by mass: 10% to 50% acetylcysteine, 1% to 10% metal chelating agent, 2% to 12% co-solvent, 0.001% to 0.01% antioxidant, 0.1% to 2% flavoring agent and 0.001% to 0.1% preservative, with the remainder being filler.

[0034] The acetylcysteine ​​soluble powder has good stability and can be put into normal daily drinking water for administration. The content of acetylcysteine ​​can be, but is not limited to, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, The content of the metal complexing agent may be, but is not limited to, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10%, the content of the cosolvent may be, but is not limited to, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11% or 12%, the content of the antioxidant may be, but is not limited to, The content of the flavoring agent may be, but is not limited to, 0.1%, 0.3%, 0.5%, 0.7%, 0.9%, 1.1%, 1.3%, 1.5%, 1.7%, 1.9% or 2%. The content of the preservative may be, but is not limited to, 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009% or 0.01%. The content of the flavoring agent may be, but is not limited to, 0.1%, 0.3%, 0.5%, 0.7%, 0.9%, 1.1%, 1.3%, 1.5%, 1.7%, 1.9% or 2%. The amount may be, but is not limited to, 0.001%, 0.005%, 0.01%, 0.015%, 0.02%, 0.025%, 0.03%, 0.035%, 0.04%, 0.045%, 0.05%, 0.055%, 0.06%, 0.065%, 0.07%, 0.075%, 0.08%, 0.085%, 0.09%, 0.095% or 0.1%.

[0035] In some embodiments, the metal complexing agent is disodium ethylenediaminetetraacetate.

[0036] In some embodiments, the co-solvent is at least one of anhydrous citric acid, polyethylene glycol, sodium xylene sulfonate, sodium lauryl sulfate, PEG6000, PVPK30, sodium citrate, and poloxamer 188, preferably sodium xylene sulfonate.

[0037] In some embodiments, the antioxidant is L-ascorbic acid.

[0038] In some embodiments, the flavoring agent is one of vanillin, cream flavor, and lemon flavor, preferably vanillin.

[0039] In some embodiments, the preservative is sorbic acid.

[0040] In some embodiments, the filler is one of anhydrous glucose, sucrose, and anhydrous lactose, preferably anhydrous lactose.

[0041] The experimental results show that the combined effect of various components makes the product acetylcysteine ​​soluble powder stable and enables acetylcysteine ​​to effectively play an auxiliary therapeutic role. It can not only significantly improve the therapeutic effect of antibiotics on chicken mycoplasma disease, but also reduce the occurrence of pathogen resistance. It can also reduce the viscosity of respiratory secretions and promote sputum discharge, and has a better clinical effect on chicken mycoplasma disease. In addition, it can also be used as an auxiliary for chicken respiratory diseases caused by other causes, and has a better clinical effect on respiratory diseases caused by various pathogens.

[0042] Acetylcysteine ​​soluble powder is in the form of a powder, and the preparation method is as follows:

[0043] (1) Prepare the raw materials and auxiliary materials according to the following weight proportions: per 100 g of soluble powder, 10-50 g of acetylcysteine, 1-10 g of metal complexing agent, 2-12 g of cosolvent, 0.1-2 g of antioxidant, 0.1-2 g of flavoring agent, 0.01-0.1 g of preservative, and the rest is anhydrous lactose.

[0044] (2) adding acetylcysteine, a metal complexing agent, and a cosolvent in a mixer in sequence, stirring to mix them evenly, to obtain an intermediate 1;

[0045] (3) first premixing an antioxidant, a flavoring agent, a preservative and a portion of anhydrous lactose, and then mixing the premix with the remaining anhydrous lactose by a doubling method to obtain an intermediate 2;

[0046] (4) Add intermediate 2 into a mixer and mix for 30 minutes until uniform, thereby obtaining acetylcysteine ​​soluble powder.

[0047] The acetylcysteine ​​soluble powder of the present application can be used to prepare drugs for the auxiliary treatment of mycoplasma disease in chickens or drugs for the auxiliary treatment of respiratory diseases in chickens, including infectious bronchitis, avian influenza or air sac inflammation in chickens. Experiments show that the acetylcysteine ​​soluble powder of the present application can be used in combination with enrofloxacin or tylosin fumarate to effectively treat Mycoplasma gallisepticum infection, in combination with Shuanghuanglian oral liquid to effectively treat infectious bronchitis in chickens, in combination with tylosin tartrate to effectively treat Mycoplasma synoviae infection in chickens, in combination with Qingwen Baidu Powder to effectively treat avian influenza, and in combination with doxycycline hydrochloride to effectively treat air sac inflammation in chickens caused by Escherichia coli.

[0048] In some embodiments, the amount of acetylcysteine ​​soluble powder is 20 mg / kg body weight as acetylcysteine.

[0049] The present application is further described below by way of examples. Unless otherwise specified, the materials in the examples are prepared according to existing methods or purchased directly from the market.

[0050] Example 1 Test on the effect of acetylcysteine ​​on improving the sensitivity of Mycoplasma gallinarum to antibiotics

[0051] 1.1 Test of acetylcysteine ​​to improve the sensitivity of Mycoplasma gallisepticum to antibiotics

[0052] In order to investigate the effect of acetylcysteine ​​on the activity of antibiotics against Mycoplasma gallisepticum and verify whether it can improve the sensitivity of Mycoplasma gallisepticum to antibiotics, the present embodiment is carried out with reference to CLSIM43-A-2011, and Mycoplasma gallisepticum and common antibiotics (including enrofloxacin, doxycycline, tylosin, tylosin) are selected to carry out the verification of this embodiment. The Mycoplasma gallisepticum used in the experiment originated from the tracheal tissue of dead chickens in chicken farms with multiple outbreaks of Mycoplasma gallisepticum disease, and was identified as Mycoplasma gallisepticum by molecular biology after separation and culture. The specific method of MIC determination is as follows.

[0053] (1) Culture medium preparation: The test used Mycoplasma PPLO broth culture medium, which was prepared according to the product instructions.

[0054] (2) Preparation of drug plates: Calculate and weigh the appropriate amount of antibiotics, prepare a stock solution of 2560 μg / ml, dilute it 10 times with culture medium to obtain a 256 μg / ml drug solution, and then dilute it twice to 0.015625 μg / ml. Take 100 μl of the diluted drug solution and add it to wells 1-9 of a 96-well plate. The drug concentrations in each well are 4 μg / ml, 2 μg / ml, 1 μg / ml, 0.5 μg / ml, 0.25 μg / ml, 0.125 μg / ml, 0.0625 μg / ml, 0.03125 μg / ml, and 0.015625 μg / ml, respectively. Add 100 μl of culture medium to wells 10-11.

[0055] (3) Preparation of mycoplasma culture: Mycoplasma counts were performed using the color change unit method (CCU), and the revived mycoplasma culture was diluted to (10 4 -10 5 )CCU / ml for standby use.

[0056] (4) Pipette 100 μl of the prepared mycoplasma culture and add it to wells 1-10 of the drug plate respectively. Well 10 is used as a bacterial solution control, 100 μl of culture medium is added to well 11, and 200 μl of pH = 6.8 culture medium (containing phenol red) is added to well 12 as a color control.

[0057] (5) After mixing, the 96-well plate was placed in a 37°C incubator with 5% CO2 and cultured for 2-3 days. The color changes of the drug-containing wells and the control wells were compared, and the lowest drug concentration that could inhibit the growth of mycoplasma (color change) was defined as MIC.

[0058] The minimum inhibitory concentration (MIC) test of the above antibiotics against Mycoplasma gallisepticum was carried out using culture media containing 0 μg / ml, 0.5 μg / ml, 1 μg / ml, 2 μg / ml and 4 μg / ml of acetylcysteine, respectively. The results are shown in Tables 1-4.

[0059] Table 1 Experimental results of the effect of different concentrations of acetylcysteine ​​on the activity of enrofloxacin against Mycoplasma gallisepticum

[0060] Table 2 Experimental results of the effect of different concentrations of acetylcysteine ​​on the activity of doxycycline against Mycoplasma gallisepticum

[0061] Table 3 Experimental results of the effect of different concentrations of acetylcysteine ​​on the activity of tylosin against Mycoplasma gallisepticum

[0062] Table 4 Experimental results of the effect of different concentrations of acetylcysteine ​​on the activity of tylosin against Mycoplasma gallisepticum

[0063] The test results showed that when the culture medium contained 1 μg / ml of acetylcysteine, the antibacterial activity of enrofloxacin, doxycycline, tiamulin and tylosin against Mycoplasma gallisepticum began to increase. When the concentration of acetylcysteine ​​increased, the antibacterial activity of the four antibiotics also increased. This result indicates that the antibacterial activity of enrofloxacin, doxycycline, tiamulin and tylosin against Mycoplasma gallisepticum is dose-related with acetylcysteine. Therefore, acetylcysteine ​​can regulate the sensitivity of Mycoplasma gallisepticum to enrofloxacin, doxycycline, tiamulin and tylosin.

[0064] 1.2 Test of acetylcysteine ​​to improve the sensitivity of Mycoplasma synoviae to doxycycline

[0065] In order to investigate the effect of acetylcysteine ​​on the activity of antibiotics against Mycoplasma gallinarum and verify whether it can improve the sensitivity of Mycoplasma gallinarum to antibiotics, the present example was carried out with reference to CLSIM43-A-2011, and Mycoplasma gallinarum and common antibiotics (including enrofloxacin, doxycycline, tylosin, tylosin) were selected to carry out the verification of this example. The Mycoplasma gallinarum used in the experiment originated from the foot pads, joints and other tissues of dead chickens in chicken farms with multiple outbreaks of Mycoplasma synoviae disease, and was isolated and cultured and identified as Mycoplasma gallinarum by molecular biology. The specific method of MIC determination is as follows.

[0066] (1) Culture medium preparation: The test used Mycoplasma PPLO broth culture medium, which was prepared according to the product instructions.

[0067] (2) Preparation of drug plates: Calculate and weigh the appropriate amount of antibiotics, prepare a stock solution of 2560 μg / ml, dilute it 10 times with culture medium to obtain a 256 μg / ml drug solution, and then dilute it twice to 0.015625 μg / ml. Take 100 μl of the diluted drug solution and add it to wells 1-9 of a 96-well plate. The drug concentrations in each well are 4 μg / ml, 2 μg / ml, 1 μg / ml, 0.5 μg / ml, 0.25 μg / ml, 0.125 μg / ml, 0.0625 μg / ml, 0.03125 μg / ml, and 0.015625 μg / ml, respectively. Add 100 μl of culture medium to wells 10-11.

[0068] (3) Preparation of mycoplasma culture: Mycoplasma counts were performed using the color change unit method (CCU), and the revived mycoplasma culture was diluted to (10 4 -10 5 )CCU / ml for standby use.

[0069] (4) Pipette 100 μl of the prepared mycoplasma culture and add it to wells 1-10 of the drug plate respectively. Well 10 is used as a bacterial solution control, 100 μl of culture medium is added to well 11, and 200 μl of pH = 6.8 culture medium (containing phenol red) is added to well 12 as a color control.

[0070] (5) After mixing, the 96-well plate was placed in a 37°C incubator with 5% CO2 and cultured for 2-3 days. The color changes of the drug-containing wells and the control wells were compared, and the lowest drug concentration that could inhibit the growth of mycoplasma (color change) was defined as MIC.

[0071] The minimum inhibitory concentration (MIC) test of the above antibiotics against Mycoplasma synoviae was carried out using culture media containing 0 μg / ml, 0.5 μg / ml, 1 μg / ml, 2 μg / ml and 4 μg / ml of acetylcysteine, respectively. The results are shown in Tables 5-8.

[0072] Table 5 Experimental results of the effect of different concentrations of acetylcysteine ​​on the activity of enrofloxacin against Mycoplasma synoviae

[0073] Table 6 Experimental results of the effect of different concentrations of acetylcysteine ​​on the activity of doxycycline against Mycoplasma synoviae

[0074] Table 7 Experimental results of the effect of different concentrations of acetylcysteine ​​on the activity of tylosin against Mycoplasma synoviae

[0075] Table 8 Experimental results of the effect of different concentrations of acetylcysteine ​​on the activity of tylosin against Mycoplasma synoviae

[0076] The test results showed that when the culture medium contained 1 μg / ml of acetylcysteine, the antibacterial activity of enrofloxacin, doxycycline, tiamulin and tylosin against Mycoplasma synoviae began to increase. When the concentration of acetylcysteine ​​increased, the antibacterial activity of the four antibiotics also increased. This result indicates that the antibacterial activity of enrofloxacin, doxycycline, tiamulin and tylosin against Mycoplasma synoviae is dose-related with acetylcysteine. Therefore, acetylcysteine ​​can regulate the sensitivity of Mycoplasma synoviae to enrofloxacin, doxycycline, tiamulin and tylosin.

[0077] Example Preparation of diacetylcysteine ​​soluble powder

[0078] This example is to determine the components of each auxiliary material in acetylcysteine ​​soluble powder, and then determine the dosage of each component.

[0079] Each component was designed according to Table 2 and packaged in aluminum foil bags, 100 g / bag. Then, the packaged components were placed at 4500±500Lx, 40°C, and 25°C, relative humidity (RH) 75±5% for 10 days in accordance with the "Guidelines for Stability Studies of Veterinary Chemical Drugs". Samples were taken on the 5th and 10th days to test the content, related substances, properties, drying loss, pH and other indicators of each component. The influencing factors investigation tests are shown in Tables 9 to 12.

[0080] Table 9 Ratio of ingredients in acetylcysteine ​​soluble powder

[0081] Table 10 Test results of each component under high temperature conditions (40°C)

[0082] Table 11 Test results of each component under high humidity conditions (25°C, 75±5%)

[0083] Table 12 Test results of each component under strong light irradiation conditions (4500±500Lx)

[0084] Acetylcysteine ​​is easily oxidized, and adding antioxidants when preparing acetylcysteine-related preparations can increase product stability. From the test results of Tables 3-5, it can be seen that after the component 6 sample was placed under high temperature, light and high humidity conditions for 10 days, its related substance indicators all exceeded the limited range (<1.5%), and the properties changed significantly, from white to light yellow powder to yellow powder, and agglomeration occurred under the two test conditions of high temperature and high humidity. Except for component 6, the indicators of the remaining components were almost unchanged after being placed under high temperature, light and high humidity conditions for 10 days, and the content and related substances met the requirements of the "Guidelines for Stability Research of Veterinary Chemical Drugs". It can be seen that the addition of antioxidants can improve product stability.

[0085] Example 3: The auxiliary therapeutic effect of acetylcysteine ​​soluble powder on chicken respiratory disease

[0086] The acetylcysteine ​​soluble powder provided in this application is mainly used for the auxiliary treatment of chicken mycoplasma disease, and can also be used for the auxiliary treatment of chicken respiratory diseases caused by other causes. In view of the above application scenarios, clinical effect verification tests of the products obtained in this application were carried out in multiple broiler farms in Henan Province where chicken respiratory diseases broke out.

[0087] 3.1 Verification of the adjuvant therapeutic effect of acetylcysteine ​​soluble powder on Mycoplasma gallisepticum

[0088] Test drug: Component 2.

[0089] Experimental location: a broiler farm in Luoyang.

[0090] A chicken respiratory disease broke out in a broiler farm in Luoyang. There were 4 chicken houses in the farm, all of which had chicken respiratory disease, and the incidence was similar. The main clinical symptoms of the chickens were rales, nose shaking, coughing, mouth breathing, etc., accompanied by an increase in the number of dead chickens. The dead chickens and dying chickens were dissected and dissected, and symptoms such as bronchial obstruction, air sac turbidity, and tracheal hemorrhage appeared. The tracheal tissue was taken for molecular identification, and the result was positive for Mycoplasma gallisepticum. It was determined that the cause of the disease in the chicken farm was Mycoplasma gallisepticum infection. The three houses with the most similar incidence were selected for the experiment, and the centralized drinking water administration method was used for large-scale group administration. 60 chickens were selected from each house for individual observation. Among them, the first house was fed with enrofloxacin solution (10%, commercially available product) according to 0.75ml / L water chickens, once a day, and the administration was continued for 3 days. The second building was fed with enrofloxacin solution (0.45 ml / L water) and acetylcysteine ​​soluble powder (20 mg / kg body weight in terms of acetylcysteine), enrofloxacin solution once a day for 3 consecutive days, and acetylcysteine ​​soluble powder once a day for 5 consecutive days. 1,000 chickens were randomly selected in the third building and fed with acetylcysteine ​​soluble powder (20 mg / kg body weight in terms of acetylcysteine) once a day for 5 consecutive days. The number of deaths of chickens in the three chicken houses was recorded daily, and the selected chickens were observed one by one before administration, 3 days after administration, and 5 days after administration, and the clinical symptom scores of each chicken were recorded according to Table 13. The summary results of the deaths of chickens in the three chicken houses after the end of the experiment are shown in Table 14, and the clinical symptom scores of individual chickens are shown in Table 15.

[0091] Table 13 Clinical symptom scoring criteria grade symptom score 0 No abnormal clinical manifestations were found 0 points 1 Chickens show mild respiratory symptoms 1 point 2 Chickens showing moderate respiratory symptoms 2 points 3 Chickens show severe respiratory symptoms 3 points 4 Chickens develop severe respiratory disease 4 points

[0092] Table 14 Summary of the total mortality of chickens in the two chicken houses after the end of the experiment

[0093] Table 15 Individual observation of clinical symptom scores of chickens

[0094] From the results in Table 14, it can be seen that the mortality rate of the house using enrofloxacin solution was 5.55%, the mortality rate of the house using enrofloxacin solution + acetylcysteine ​​soluble powder was 2.19%, and the mortality rate of the house using acetylcysteine ​​soluble powder was 6.90%; the mortality rate of the house using acetylcysteine ​​soluble powder alone did not decrease; compared with the house using enrofloxacin solution alone, the mortality rate of the chicken flock in the house using enrofloxacin solution + acetylcysteine ​​soluble powder was significantly reduced; therefore, the use of acetylcysteine ​​soluble powder of component 2 in combination with enrofloxacin solution can significantly reduce the mortality rate of Mycoplasma gallisepticum disease.

[0095] From the results in Table 15, it can be seen that the use of acetylcysteine ​​soluble powder can significantly reduce the clinical symptom scores of diseased chickens, indicating that adjuvant treatment with acetylcysteine ​​soluble powder of component 2 while administering enrofloxacin solution can improve the clinical therapeutic effect of enrofloxacin on Mycoplasma gallisepticum.

[0096] 3.2 Verification of the auxiliary therapeutic effect of acetylcysteine ​​soluble powder on infectious bronchitis in chickens

[0097] Test drug: Component 1.

[0098] Experimental location: a broiler farm in Nanyang.

[0099] A broiler farm in Nanyang had an outbreak of chicken respiratory disease during the season transition. The farm had three chicken houses, all of which had chicken respiratory disease, and the incidence was similar. The main clinical symptoms of the chickens were rales, nose shaking, coughing, mouth breathing, strange cries, etc., accompanied by an increase in the number of dead chickens. The dead chickens and dying chickens were dissected and dissected, showing symptoms such as bronchial obstruction, air sac turbidity, and tracheal hemorrhage. Tracheal tissue was taken for molecular identification, and the results showed that the infectious bronchitis virus was positive, and the chicken farm was determined to be infectious bronchitis. At the same time, three chicken houses were selected for the experiment, and large-scale drug administration was carried out by centralized drinking water administration. 60 chickens were selected from each house for individual observation. Among them, the first house was given Shuanghuanglian oral liquid (commercially available product) at 0.5ml / kg body weight, once a day, for 5 consecutive days. The second building was fed with Shuanghuanglian oral liquid (0.3ml / kg) and acetylcysteine ​​soluble powder (20mg / kg body weight in terms of acetylcysteine) once a day for 5 consecutive days. 1,000 chickens were randomly selected in the third building and fed with acetylcysteine ​​soluble powder (20mg / kg body weight in terms of acetylcysteine) once a day for 5 consecutive days. The number of deaths of chickens in the three chicken houses was recorded daily, and the selected chickens were observed one by one before administration, 3 days after administration, and 5 days after administration, and the clinical symptom scores of each chicken were recorded according to Table 13. The summary results of the deaths of chickens in the three chicken houses after the end of the experiment are shown in Table 16, and the clinical symptom scores of individual chickens are shown in Table 17.

[0100] Table 16 Summary of the total mortality of chickens in the two chicken houses after the end of the experiment

[0101] Table 17 Individual observation of clinical symptom scores of chickens Group Before administration Drug administration 3d 5 days of medication Building 1 3.58±0.72 1.98±0.95 0.57±0.77 Building 2 3.50±0.79 0.82±1.18 0.45±0.83 Building 3 3.63±0.66 2.75±0.97 1.05±1.03

[0102] From the results in Table 16, it can be seen that the mortality rate of the house using Shuanghuanglian oral liquid was 4.84%, the mortality rate of the house using Shuanghuanglian oral liquid + acetylcysteine ​​soluble powder was 1.98%, and the mortality rate of the house using acetylcysteine ​​soluble powder was 7.10%; the mortality rate of the house using acetylcysteine ​​soluble powder alone did not decrease; compared with the house using Shuanghuanglian oral liquid alone, the mortality rate of the chicken flock using Shuanghuanglian oral liquid + acetylcysteine ​​soluble powder was significantly reduced; therefore, the use of acetylcysteine ​​soluble powder of component 1 in combination with Shuanghuanglian oral liquid can significantly reduce the mortality rate of infectious bronchitis in chickens.

[0103] From the results in Table 17, it can be seen that the use of acetylcysteine ​​soluble powder can significantly reduce the clinical symptom scores of diseased chickens, indicating that the auxiliary treatment of acetylcysteine ​​soluble powder of component 1 while administering Shuanghuanglian oral liquid can improve the clinical therapeutic effect of Shuanghuanglian oral liquid on infectious bronchitis in chickens.

[0104] 3.3 Verification of the auxiliary therapeutic effect of acetylcysteine ​​soluble powder on chicken synoviae mycoplasma disease

[0105] Test drug: Component 3.

[0106] Experimental location: an egg-laying chicken farm in Luoyang.

[0107] A laying hen farm in Luoyang has four chicken houses, each of which houses 25,000 chickens. At 56 days of age, the chickens in the four chicken houses showed clinical symptoms such as decreased appetite, poor development, low uniformity, and small body weight of sick chickens. Autopsy of dead chickens showed purulent secretions in swollen joints and paw pads. The joint secretions were taken for molecular identification, and the results showed that Mycoplasma synoviae was positive. Three of the chicken houses were selected for the experiment. The first house was given tylosin tartrate soluble powder (100g: 10g, commercially available product) at a dose of 5g / L water, once a day for 5 consecutive days. The second house was fed with tylosin tartrate soluble powder (3g / L water) and acetylcysteine ​​soluble powder (calculated as acetylcysteine, 20mg / kg body weight), once a day for 5 consecutive days. 1000 chickens were randomly selected from the third house and fed with acetylcysteine ​​soluble powder (20 mg / kg body weight as acetylcysteine) once a day for 5 consecutive days. The number of dead chickens and feed intake in the three chicken houses were observed from before administration to 5 days after administration, and 100 chickens were randomly selected from each of the three chicken houses before administration and 5 days after administration, and the mortality rate, feed intake and individual weight of chickens in the three chicken houses were compared. The specific results are shown in Tables 18-19.

[0108] Table 18 Summary of total mortality of chickens in the two chicken houses during the trial period

[0109] Table 19 Observation results of feed intake and body weight of chickens

[0110] From the results in Table 18, it can be seen that the mortality rate of the houses using tylosin tartrate soluble powder was 4.07%, the mortality rate of the houses using tylosin tartrate soluble powder + acetylcysteine ​​soluble powder was 1.01%, and the mortality rate of the houses using acetylcysteine ​​soluble powder was 6.50%; the mortality rate of the houses using acetylcysteine ​​soluble powder alone did not decrease; compared with the houses using tylosin tartrate soluble powder alone, the mortality rate of the chickens using tylosin tartrate soluble powder + acetylcysteine ​​soluble powder was significantly reduced, so the use of acetylcysteine ​​soluble powder of component 3 in combination with tylosin tartrate soluble powder can significantly reduce the mortality rate of infectious bronchitis in chickens.

[0111] From the results in Table 19, it can be seen that the use of acetylcysteine ​​soluble powder can significantly increase the feed intake and body weight of the chickens, indicating that the adjuvant treatment of acetylcysteine ​​soluble powder of component 3 while administering tylosin tartrate soluble powder can improve the clinical therapeutic effect of tylosin tartrate soluble powder on chicken synoviae mycoplasma disease.

[0112] 3.4 Verification of the adjuvant therapeutic effect of acetylcysteine ​​soluble powder on Mycoplasma gallisepticum

[0113] Test drug: Component 4.

[0114] Experimental location: a broiler farm in Kaifeng.

[0115] A chicken respiratory disease broke out in a broiler farm in Kaifeng. There were 3 chicken houses in the farm, all of which had chicken respiratory disease at 18 days of age, and the incidence was similar. The main clinical symptoms of the chickens were rales, nose shaking, coughing, mouth breathing, etc., accompanied by an increase in the number of dead chickens. The dead chickens and dying chickens were dissected and dissected, and symptoms such as bronchial obstruction, air sac turbidity, and tracheal hemorrhage appeared. The tracheal tissue was taken for molecular identification, and the result was positive for Mycoplasma gallisepticum. It was determined that the cause of the disease in the chicken farm was Mycoplasma gallisepticum infection. These 3 chicken houses were selected for the experiment, and the centralized drinking water administration method was used for large-scale group administration. 60 chickens were selected from each house for individual observation. Among them, the first house was fed with 1.25g / L water of tiamulin soluble powder (10%, commercially available), once a day, for 3 consecutive days. The second building was fed with tiamulin fumarate soluble powder (0.85g / L water) and acetylcysteine ​​soluble powder (20mg / kg body weight in terms of acetylcysteine). The tiamulin fumarate soluble powder was administered once a day for 3 consecutive days, and the acetylcysteine ​​soluble powder was administered once a day for 5 consecutive days. 1,000 chickens were randomly selected in the third building and fed with acetylcysteine ​​soluble powder (20mg / kg body weight in terms of acetylcysteine) once a day for 5 consecutive days. The number of deaths of chickens in the three chicken houses was recorded daily, and the selected chickens were observed one by one before administration, 3 days after administration, and 5 days after administration, and the clinical symptom scores of each chicken were recorded according to Table 13. The summary results of the deaths of chickens in the three chicken houses after the end of the experiment are shown in Table 20, and the clinical symptom scores of individual chickens are shown in Table 21.

[0116] Table 20 Summary of total chicken deaths in the two chicken houses after the end of the experiment

[0117] Table 21 Individual observation of clinical symptom scores of chickens Group Before administration Drug administration 3d 5 days of medication Building 1 3.25±0.89 1.98±1.05 0.62±0.80 Building 2 3.23±0.91 0.82±1.21 0.48±0.85 Building 3 3.17±0.91 2.60±1.28 0.70±0.94

[0118] From the results in Table 20, it can be seen that the mortality rate of the house using tiamulin fumarate soluble powder was 4.73%, the mortality rate of the house using tiamulin fumarate soluble powder + acetylcysteine ​​soluble powder was 1.47%, and the mortality rate of the house using acetylcysteine ​​soluble powder was 6.40%; the mortality rate of the house using acetylcysteine ​​soluble powder alone did not decrease; compared with the house using tiamulin fumarate soluble powder alone, the mortality rate of the chickens using tiamulin fumarate soluble powder + acetylcysteine ​​soluble powder was significantly reduced. Therefore, the combined use of acetylcysteine ​​soluble powder of component 4 and tiamulin fumarate soluble powder can significantly reduce the mortality rate of Mycoplasma gallisepticum disease.

[0119] From the results in Table 21, it can be seen that the use of acetylcysteine ​​soluble powder can significantly reduce the clinical symptom scores of diseased chickens, indicating that the administration of acetylcysteine ​​soluble powder of fumarate and component 4 for auxiliary treatment can improve the clinical therapeutic effect of acetylcysteine ​​soluble powder of fumarate on Mycoplasma gallisepticum.

[0120] 3.5 Verification of the auxiliary therapeutic effect of acetylcysteine ​​soluble powder on avian influenza

[0121] Test drug: Component 5.

[0122] Experimental location: an egg-laying chicken farm in Luoyang.

[0123] There are three chicken houses in a laying hen farm in Luoyang, each of which houses 30,000 chickens. The chickens in the three chicken houses showed depression, fever, loss or disappearance of appetite, decreased egg production, and moderate respiratory symptoms at 28 weeks of age. The sick chickens had swollen heads, coughing, shaking noses, tracheal rales, and a lot of tears. The autopsy of dead chickens showed mucus and cheese in the trachea, severe tracheal bleeding, multiple intestinal bleeding, and turbidity of abdominal air sacs. The trachea and lungs were taken for molecular identification, and the results showed that the avian influenza H9 was positive. These three chicken houses were selected for the experiment, and large groups were administered by centralized drinking water. Among them, the first house was given Qingwen Baidu San (commercially available product) at 2g / kg feed, once a day, for 5 consecutive days. The second house was fed Qingwen Baidu San (1g / kg feed) and acetylcysteine ​​soluble powder (20mg / kg body weight in terms of acetylcysteine), once a day, for 5 consecutive days. 1,000 chickens were randomly selected from the third house and fed with acetylcysteine ​​soluble powder (20 mg / kg body weight in terms of acetylcysteine) once a day for 5 consecutive days. The number of deaths and feed intake of chickens in the three houses were recorded daily. 30 sick chickens with typical symptoms were selected from each house, and their body temperatures were observed and measured before, 3 days after, and 5 days after administration. The summary results of the deaths of chickens in the three houses after the end of the experiment are shown in Table 22, the statistical results of feed intake and egg production are shown in Table 23, and the results of body temperature measurement are shown in Table 24.

[0124] Table 22 Summary of total chicken deaths in the two chicken houses after the end of the experiment

[0125] Table 23 Observation results of feed intake and body weight of chickens

[0126] Table 24 Observation results of feed intake and body weight of chickens

[0127] From the results in Table 22, it can be seen that the mortality rate of the house using Qingwen Baidu San was 4.60%, the mortality rate of the house using Qingwen Baidu San + acetylcysteine ​​soluble powder was 2.33%, and the mortality rate of the house using acetylcysteine ​​soluble powder was 6.30%; the mortality rate of the house using acetylcysteine ​​soluble powder alone did not decrease; compared with the house using Qingwen Baidu San alone, the mortality rate of the chicken flock using Qingwen Baidu San + acetylcysteine ​​soluble powder was significantly reduced. Therefore, the combined use of acetylcysteine ​​soluble powder of component 5 and Qingwen Baidu San can significantly reduce the mortality rate of avian influenza.

[0128] From the results in Tables 23 and 24, it can be seen that the use of acetylcysteine ​​soluble powder can significantly increase the feed intake and egg production rate of the chickens, and can significantly improve the fever symptoms of the chickens, indicating that giving the chickens Qingwen Baidu Powder and using acetylcysteine ​​soluble powder of component 5 for auxiliary treatment can improve the clinical treatment effect of Qingwen Baidu Powder on avian influenza.

[0129] 3.6 Verification of the auxiliary therapeutic effect of acetylcysteine ​​soluble powder on chicken air sacculitis

[0130] Test drug: Component 2.

[0131] Experimental location: a broiler farm in Luoyang.

[0132] A broiler farm in Luoyang has three chicken houses, each of which raises 20,000 chickens. The chickens in the three chicken houses showed clinical symptoms such as depression and fatigue at 20 days of age, accompanied by respiratory symptoms such as rales and coughs. Autopsy showed that the air sacs were turbid with yellow cheesy substances, and fibrinous exudates were seen on the heart and liver. The air sac, heart and liver tissues were taken for bacterial isolation and culture. The results showed that the chickens were infected with Escherichia coli. According to the results of the in vitro drug sensitivity test, the pathogenic bacteria were sensitive to doxycycline. At the same time, three chicken houses were selected for the experiment, and large-scale drug administration was carried out by centralized drinking water administration. Among them, the first house was given doxycycline hydrochloride soluble powder (20%, commercially available product) at 1.5g / L water, once a day, for 5 consecutive days. The second house was fed with doxycycline hydrochloride soluble powder (1.0g / L water) and acetylcysteine ​​soluble powder (calculated as acetylcysteine, 20mg / kg body weight), once a day, for 5 consecutive days. 1,000 chickens were randomly selected from the third house and fed with acetylcysteine ​​soluble powder (20 mg / kg body weight as acetylcysteine) once a day for 5 consecutive days. The number of deaths of chickens in the three houses was recorded daily. The summary results of the deaths of chickens in the three houses after the end of the experiment are shown in Table 25.

[0133] Table 25 Summary of the mortality of chickens in the two chicken houses after the end of the experiment

[0134] From the results in Table 25, it can be seen that the mortality rate of the house using doxycycline hydrochloride soluble powder was 4.84%, the mortality rate of the house using doxycycline hydrochloride soluble powder + acetylcysteine ​​soluble powder was 272%, and the mortality rate of the house using acetylcysteine ​​soluble powder was 6.50%; the mortality rate of the house using acetylcysteine ​​soluble powder alone did not decrease; compared with the house using doxycycline hydrochloride soluble powder alone, the mortality rate of the chickens using doxycycline hydrochloride soluble powder + acetylcysteine ​​soluble powder was significantly reduced, and the clinical symptoms were significantly improved. Therefore, the use of acetylcysteine ​​soluble powder of component 2 can significantly reduce the mortality rate of chicken Escherichia coli air sacculitis, indicating that the clinical treatment of doxycycline hydrochloride soluble powder on chicken infectious bronchitis can be improved by giving doxycycline hydrochloride soluble powder and using acetylcysteine ​​soluble powder of component 2 for auxiliary treatment.

[0135] Note that the above are only preferred embodiments of the present application and the technical principles used. Those skilled in the art will understand that the present application is not limited to the specific embodiments described herein, and that various obvious changes, readjustments and substitutions can be made by those skilled in the art without departing from the protection scope of the present application. Therefore, although the present application is described in more detail through the above embodiments, the present application is not limited to the above embodiments, and may also include more other equivalent embodiments without departing from the technical concept of the present application, all of which belong to the protection scope of the present application.

Claims

1. Application of acetylcysteine ​​in the preparation of drugs for improving the antibiotic sensitivity of chicken mycoplasma.

2. The use according to claim 1, characterized in that: The antibiotics include at least one of tiamulin, doxycycline, tylosin or enrofloxacin.

3. The use according to claim 2, characterized in that: The content of acetylcysteine ​​is not less than 1 μg / ml, preferably (1 to 4) μg / ml.

4. The use according to claim 1, characterized in that: Mycoplasma galli includes Mycoplasma gallisepticum and Mycoplasma synoviae.

5. An acetylcysteine ​​soluble powder, characterized in that, The invention is composed of the following components by mass percentage: 10% to 50% acetylcysteine, 1% to 10% metal complexing agent, 2% to 12% co-solvent, 0.001% to 0.01% antioxidant, 0.1% to 2% flavoring agent and 0.001% to 0.1% preservative, and the balance is filler.

6. Acetylcysteine ​​soluble powder according to claim 4, characterized in that, The composition is composed of the following components by mass percentage: 15% to 30% acetylcysteine, 2% to 6% metal complexing agent, 3% to 8% cosolvent, 0.002% to 0.006% antioxidant, 0.2% to 1% flavoring agent and 0.005% to 0.05% preservative, and the balance is filler; Preferably, the composition is composed of the following components by mass percentage: 20% acetylcysteine, 4% metal complexing agent, 5% cosolvent, 0.004% antioxidant, 0.5% flavoring agent and 0.01% preservative, and the balance is filler.

7. The acetylcysteine ​​soluble powder according to claim 5, characterized in that The metal complexing agent is disodium ethylenediaminetetraacetate; and / or, The cosolvent is at least one of anhydrous citric acid, polyethylene glycol, sodium xylene sulfonate, sodium lauryl sulfate, PEG6000, PVPK30, sodium citrate, and poloxamer 188, preferably sodium xylene sulfonate; and / or, The antioxidant is L-ascorbic acid; and / or, The flavoring agent is one of vanillin, cream flavor, and lemon flavor, preferably vanillin; and / or, The preservative is sorbic acid; and / or, The filler is one of anhydrous glucose, sucrose and anhydrous lactose, preferably anhydrous lactose.

8. The method for preparing the acetylcysteine ​​soluble powder according to any one of claims 5 to 7, characterized in that: The following steps are involved: Acetylcysteine, a metal complexing agent and a cosolvent are uniformly mixed to obtain an intermediate 1; The antioxidant, the flavoring agent, the preservative and a part of the anhydrous lactose are premixed, and then the premix is ​​mixed evenly with the remaining anhydrous lactose by a doubling method to obtain an intermediate 2; The intermediate 1 and the intermediate 2 are mixed evenly to obtain acetylcysteine ​​soluble powder.

9. Use of the acetylcysteine ​​soluble powder according to any one of claims 5 to 7 in the preparation of a medicament for the auxiliary treatment of chicken mycoplasma disease.

10. Use of the acetylcysteine ​​soluble powder according to any one of claims 5 to 7 in the preparation of a drug for auxiliary treatment of chicken respiratory diseases; Respiratory diseases in chickens include infectious bronchitis, avian influenza or air sac disease.

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