Application of Taxifolin in the preparation of drugs against Mycoplasma gallisepticum and Mycoplasma synoviae

Abstract

The present invention belongs to the technical field of veterinary drugs, and specifically relates to the application of taxifolin in the preparation of drugs against Mycoplasma gallisepticum and Mycoplasma synoviae. The applicant has found that taxifolin has the effects of inhibiting the growth of Mycoplasma gallisepticum and Mycoplasma synoviae and killing Mycoplasma gallisepticum and Mycoplasma synoviae. The minimum inhibitory concentration for Mycoplasma gallisepticum is 125 μg / mL and the minimum bactericidal concentration is 500 μg / mL; the minimum inhibitory concentration for Mycoplasma synoviae is 125 μg / mL and the minimum bactericidal concentration is 500 μg / mL. The technical solution of the present invention overcomes the toxic and side effects of vaccines, chemical drugs, and antibiotics and the hidden dangers of food safety, and has broad application prospects and development value in the fields of food processing, medicine, and aquaculture.

Description

Technical Field

[0001] The invention belongs to the technical field of veterinary drugs, and particularly relates to application of taxifolin in preparing drugs against Mycoplasma gallisepticum and Mycoplasma synoviae. Background Art

[0002] Poultry mycoplasmosis is mainly caused by Mycoplasma gallisepticum and Mycoplasma synoviae, which can be transmitted both horizontally and vertically. Infection with Mycoplasma gallisepticum can cause respiratory diseases, decreased egg production, reduced hatching rate, and abnormal eggshells in poultry. In addition to the above symptoms, infection with Mycoplasma synoviae can also cause infectious synovitis in poultry, causing huge economic losses to the poultry industry (Anneke Feberwee, Sjaak de Wit, Remco Dijkman, Clinical expression, epidemiology, and monitoring of Mycoplasma gallisepticum and Mycoplasma synoviae: an update, Avian Pathol. 2022 Feb; 51(1): 2-18.).

[0003] Mycoplasmas lack cell walls and are naturally resistant to antimicrobial drugs that target peptidoglycan, such as β-lactams, glycopeptides, and fosfomycins (Ken B Waites, Li Xiao, Yang Liu, et al. Mycoplasma pneumoniae from the Respiratory Tract and Beyond. Clin Microbiol Rev. 2017 Jul; 30(3): 747-809.). In addition, mycoplasmas lack the folate synthesis pathway and are resistant to sulfonamide antimicrobial drugs that target the folate synthesis pathway (Fabricio BM Arraes, Maria José A. de Carvalho, Andrea Q. etal.Differential metabolism of Mycoplasma species as revealed by theirgenomes.Genetics and Molecular Biology,30,1,182-189(2007);KK Schultz,E LStrait,BZ Erickson,et al.Optimization of an antibiotic sensitivity assay forMycoplasma hyosynoviae and susceptibility profiles of field isolates from1997to 2011.Vet Microbiol. 2012Jul 6;158(1-2):104-8.). In addition to the above drugs, polymyxin, nalidixic acid, rifampicin and some oxazolidinone drugs, such as linezolid, are also ineffective against mycoplasma (Ken B Waites, Li Xiao, Yang Liu, et al. Mycoplasma pneumoniae from the Respiratory Tract and Beyond. Clin Microbiol Rev. 2017 Jul; 30 (3): 747-809.). In summary, compared with bacteria, mycoplasma lacks some biosynthetic functions, and many drugs reported to be useful for bacteria or fungi are ineffective against mycoplasma, resulting in relatively few drugs that can be used to treat mycoplasma infections.

[0004] There are large differences in the drug resistance of mycoplasmas from different sources. For example, Mycoplasma synoviae from animals is seriously resistant to macrolides and fluoroquinolones (Zhang Huiru, Xiao Xia, Yang Tong, et al. Study on the resistance of Mycoplasma synoviae to commonly used antibiotics [J]. China Poultry. 2021, 43(04)); Genital Mycoplasmas from humans are resistant to quinolones and aminoglycosides to varying degrees, but are highly sensitive to macrolides and tetracyclines (Wei Jingwen, Liu Enling, Liu Zhihui, et al. Analysis of 1006 cases of female genital mycoplasma infection and drug resistance [J]. Journal of North China University of Technology (Medical Edition). 2020, 22(02)).

[0005] Antibiotic treatment is one of the most effective strategies for controlling avian mycoplasma infection. However, there are relatively few drugs that can be used to treat mycoplasma infection and mycoplasmas from different sources have varying resistance to antimicrobial drugs. Therefore, it is an urgent need for the current poultry industry to find safe / efficient antibiotic alternatives to strengthen the prevention and control of poultry mycoplasma disease. Chinese herbal medicine not only has a conditioning effect on the body, but also has an anti-infection effect on pathogenic microorganisms, and is an important source for the research and development of antibiotic alternatives.

[0006] Taxifolin is a bioflavonoid extract extracted from plants such as larch, Chinese wolfberry leaves, and Chinese smilax. Studies have shown that taxifolin has a variety of biological activities such as prevention and treatment of gouty arthritis, human circulatory system diseases, and diabetes, and has low toxic side effects. Taxifolin (molecular weight: 304.25, molecular formula: C 15 H 12 O 7 ) is as follows:

[0007]

[0008] So far, there is no report on the research of Taxol in resisting mycoplasma. Summary of the invention

[0009] The technical problem to be solved by the present invention is to overcome the serious drug resistance of Mycoplasma galliscepticum and Mycoplasma synoviae, and to provide the use of dithyracin in the preparation of drugs against Mycoplasma galliscepticum and Mycoplasma synoviae. Dithyracin has good antibacterial and bactericidal effects on both mycoplasmas, and has good application prospects in the prevention and treatment of avian mycoplasma diseases.

[0010] To achieve the above object, the present invention adopts the following technical solutions:

[0011] The invention discloses an application of taxifolin in the preparation of drugs against Mycoplasma gallisepticum and Mycoplasma synoviae, including the application of taxifolin in the preparation of drugs for treating or preventing Mycoplasma gallisepticum and Mycoplasma synoviae infection, including using the substance to inhibit the growth of Mycoplasma gallisepticum and Mycoplasma synoviae in vitro, or directly killing Mycoplasma gallisepticum and Mycoplasma synoviae; or preparing a preparation for drugs for diseases infected with Mycoplasma gallisepticum and Mycoplasma synoviae.

[0012] Compared with the prior art, the present invention has the following advantages and effects:

[0013] The present invention discovers for the first time that taxifolin can be used to fight Mycoplasma gallisepticum and Mycoplasma synoviae. The present invention studies and verifies through various methods that taxifolin has a good antibacterial effect on Mycoplasma gallisepticum and Mycoplasma synoviae, laying a theoretical foundation for the development of new drugs against avian mycoplasma disease, and even providing candidate substances for the discovery of antibiotic substitutes, and has a good application prospect in the prevention and treatment of avian mycoplasma disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 This is a dynamic analysis of the growth curve of taxifolin against the growth of Mycoplasma gallisepticum.

[0015] Figure 2 This is a dynamic analysis of the growth curve of taxifolin against the growth of chicken synoviae Mycoplasma. DETAILED DESCRIPTION

[0016] The present invention is further described below in conjunction with the accompanying drawings and specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0017] Unless otherwise specified, the reagents and materials used in the following examples are commercially available. The present invention uses Mycoplasma gallisepticum and Mycoplasma synoviae in avian mycoplasmas as examples to illustrate that Taxifolin may also have inhibitory and killing effects on other avian mycoplasmas; the structural formula of Taxifolin is as follows:

[0018]

[0019] Statistical analysis of the following examples of the present invention: All experiments were repeated at least 3 times independently.

[0020] Embodiment 1:

[0021] Determination of the minimum inhibitory concentration and minimum bactericidal concentration of taxifolin against Mycoplasma gallisepticum and Mycoplasma synoviae:

[0022] 1. Materials

[0023] Modified Frey's medium is used for drug sensitivity experiments to detect the antibacterial efficacy of drugs.

[0024] 2. Test methods

[0025] The mycoplasma suspension cultured to the logarithmic growth phase was diluted to an appropriate concentration and added to sterile 96-well plates. 100 μL of the diluted mycoplasma suspension was added to wells 1 to 11 to make the final concentration of the mycoplasma suspension in each well 10 5 CCU / mL;

[0026] The Taxifolin solution was diluted in multiples with modified Frey's medium, and different concentrations of Taxifolin solution were added to the above 96-well plate. The diluted Taxifolin solution was added to wells 1 to 10, 100 μL per well, and 100 μL of modified Frey's medium was added to well 11, finally obtaining 200 μL of mycoplasma suspension as a growth control. Only 200 μL of modified Frey's medium was added to well 12 as a negative control. 37℃5% CO 2 Incubate in the incubator for 48 hours, observe the color change of the culture medium in the 96-well plate, and determine the MIC value. Take the bacterial solution from each well and spread it on the modified Frey solid medium. If the number of colonies is less than 5, it is the MBC value.

[0027] 3. Results

[0028] As shown in Table 1, when the concentration of taxifolin is 125 μg / mL, it has an inhibitory effect on the growth of Mycoplasma gallisepticum and Mycoplasma synoviae. When the concentration of taxifolin is 500 μg / mL, it has a bactericidal effect on Mycoplasma gallisepticum and Mycoplasma synoviae.

[0029] Table 1 Minimum inhibitory concentration and minimum bactericidal concentration of taxifolin against Mycoplasma gallisepticum and Mycoplasma synoviae

[0030]

[0031] Embodiment 2:

[0032] Inhibitory effect of taxifolin on the growth of Mycoplasma gallisepticum and Mycoplasma synoviae:

[0033] The revived Mycoplasma gallisepticum and Mycoplasma synoviae that reached the third generation were inoculated into fresh modified Frey's medium at a ratio of 1:10.

[0034] Taxifolin solution was added to the above mycoplasma suspension to make the final concentrations 1 / 2MIC, 1MIC, 2MIC, 4MIC, and 8MIC, respectively; the mycoplasma suspension without Taxifolin was used as a positive control, and the modified Frey's medium alone was used as a negative control;

[0035] 37℃ 5% CO 2 The cells were cultured in an incubator, and the CCU of the mycoplasma suspension at each drug concentration was determined at 0, 12, 24, 36, and 48 h, respectively, to obtain the growth curves of Mycoplasma gallisepticum and Mycoplasma synoviae at different Taxifolin concentrations.

[0036] Depend on Figure 1 , Figure 2It can be seen that when the final concentration of Taxifolin was 62.5 μg / mL, 125 μg / mL, and 250 μg / mL (i.e., 1 / 2MIC, 1MIC, and 2MIC), compared with the control group without Taxifolin, the growth of the two mycoplasmas was inhibited at each time point, and was dose-dependent with the concentration of the Taxifolin solution. When the final concentration of Taxifolin was greater than or equal to 500 μg / mL (i.e., 4MIC), both mycoplasmas were completely killed.

[0037] The above results show that taxifolin can not only inhibit the growth of Mycoplasma gallisepticum and Mycoplasma synoviae, but also kill Mycoplasma gallisepticum and Mycoplasma synoviae.

[0038] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Application of Taxifolin in the preparation of drugs against Mycoplasma gallisepticum and Mycoplasma synoviae.

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