Biological preparation for treating chicken proventriculitis and preparation method thereof

Abstract

The invention belongs to the field of biology, and discloses a preparation method of a biological agent for treating chicken proventriculitis, which comprises the following steps: carrying out anaerobic fermentation on a traditional Chinese medicine preparation by adopting enterococcus faecium to obtain the biological agent; the traditional Chinese medicine preparation consists of the following components in parts by weight: 10-20 parts of codonopsis pilosula; 5-15 parts by weight of Radix Astragali; 5-15 parts by weight of bighead atractylodes rhizome; 5-15 parts by weight of Poria cocos; 5-15 parts by weight of licorice; 5-15 parts by weight of dried orange peel; 5-15 parts by weight of mangnolia officinalis; 5-15 parts by weight of dried ginger; and 5-15 parts by weight of hawthorn. According to the method, after enterococcus faecium is adopted for fermenting the traditional Chinese medicine preparation for treating chicken proventriculitis, the prevention and treatment effect on chicken proventriculitis can be remarkably improved. Meanwhile, the invention further provides the biological preparation prepared on the basis of the preparation method.

Description

Technical Field

[0001] The invention relates to the biological field, and in particular to a biological preparation for treating chicken glandular gastritis and a preparation method thereof. Background Art

[0002] Chicken proventriculus, also known as infectious proventriculus, is a newly emerging infectious disease characterized by enlarged proventriculus. The disease is manifested by depression, reduced diet, and gradual weight loss in sick chickens. In the middle and late stages of the disease, sick chickens become extremely emaciated and pale, and eventually die of exhaustion.

[0003] The product Changweikang I sold by our company is a traditional Chinese medicine preparation specifically for poultry digestive tract diseases, and it also has a good effect on chicken glandular gastritis.

[0004] At the same time, in this field, there are also many traditional Chinese medicine prescriptions for the treatment of chicken glandular gastritis, such as publication number CN106214736A, the subject of a patent application for a traditional Chinese medicine composition for treating chicken glandular gastritis, its preparation method and application, using Elsholtzia ciliata, Isatis indigotica, Codonopsis pilosula, and Licorice to treat the disease; publication number CN105663546A, the subject of a patent application for a traditional Chinese medicine composition for treating chicken glandular gastritis, using Codonopsis pilosula, cinnamon bark, Astragalus membranaceus, Atractylodes macrocephala, chicken gizzard lining, Areca catechu, montmorillonite, Cuttlefish sheath, Cortex Moutan, and Licorice to treat the disease.

[0005] The technical problem solved in this case is: how to further improve the therapeutic effect on chicken glandular gastritis. Summary of the invention

[0006] The purpose of the present invention is to provide a method for preparing a biological preparation for treating chicken glandular gastritis. After the traditional Chinese medicine preparation for treating chicken glandular gastritis is fermented by Enterococcus faecium, the preventive and therapeutic effect on chicken glandular gastritis can be significantly improved.

[0007] At the same time, the present invention also provides a biological preparation prepared based on the preparation method.

[0008] To achieve the above objectives, this application discloses: A method for preparing a biological preparation for treating chicken glandular gastritis, wherein the biological preparation is obtained by anaerobic fermentation of a traditional Chinese medicine preparation by Enterococcus faecium; the traditional Chinese medicine preparation is composed of the following components: Codonopsis pilosula 10-20 parts by weight; 5-15 parts by weight of Astragalus; Atractylodes macrocephala 5-15 parts by weight; Poria 5-15 parts by weight; 5-15 parts by weight of liquorice; Tangerine peel 5-15 parts by weight; 5-15 parts by weight of Magnolia officinalis; 5-15 parts by weight of dried ginger; Hawthorn 5-15 parts by weight; The classification of Enterococcus faecium is named Enterococcus faecium , the deposit number is GDMCC NO: 65031; the deposit date is August 20, 2024, and it is deposited in Guangdong Provincial Microbiological Culture Collection Center, and the deposit address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0009] In the above-mentioned preparation method, the method is specifically: Step 1: Activate Enterococcus faecium to obtain a viable count of (1~10)*10 8 CFU / ml of bacterial solution; Step 2: Inoculate the bacterial liquid into the culture medium containing the traditional Chinese medicine preparation, and in the absence of oxygen, control the fermentation temperature at 30-35°C for 10-15 days to obtain the biological preparation; The culture medium is a solid culture medium, which is composed of a crushed traditional Chinese medicine preparation, a carbon source, a nitrogen source, and an inorganic salt; and the water content of the solid culture medium is controlled at 40-60wt%.

[0010] In the above preparation method, the solid culture medium comprises 100 parts by weight of the traditional Chinese medicine preparation, 5 to 10 parts by weight of a carbon source, 3 to 5 parts by weight of a nitrogen source, and 0.5 to 1 part by weight of an inorganic salt.

[0011] In the above preparation method, the carbon source is one or more of bran, rice bran, glucose, and molasses; the nitrogen source is peptone; and the inorganic salt is one or more combinations of K2HPO4, KH2PO4, MgSO4, NaCl, KCl, and FeSO4.

[0012] In the above preparation method, the weight ratio of the bacterial liquid to the culture medium is 1-5:100.

[0013] In the above preparation method, the activation method in step 1 is: inoculating Enterococcus faecium into a malt wort liquid culture medium, and culturing on a shaker at 28-30° C. for 6-10 hours.

[0014] In addition, the present invention also discloses a biological preparation for treating chicken glandular gastritis, which is prepared by any of the above methods.

[0015] This application has at least the following beneficial effects: 1. The present invention uses Enterococcus faecium to ferment the company's Changweikang I, which can significantly improve the prevention and treatment effect of Changweikang I on chicken glandular gastritis.

[0016] 2. The Enterococcus faecium in the biological preparation for treating chicken glandular gastritis of the present invention is added to the feed in a live bacterial state, and as a probiotic, it can significantly improve the prevention and treatment effect of chicken glandular gastritis.

[0017] 3. The fermentation of traditional Chinese medicine preparations and the probiotic characteristics of Enterococcus faecium itself comprehensively improve the prevention and treatment effect of chicken glandular gastritis. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a photo of a healthy chicken's gizzard, which is smooth and has no ulcers; Figure 2 This is a photo of a gizzard ulcer caused by glandular gastritis in chickens; Figure 3 This is a photo of a gizzard ulcer with bleeding caused by glandular gastritis in a chicken. DETAILED DESCRIPTION

[0019] The present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. In the description of the present invention, it should be noted that, if no specific conditions are specified in the embodiments, the conditions are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0020] The strains used in the present invention are derived from: Candida tropicalis (GDMCC 65030): Classification and naming candida tropocalis , the deposit number is GDMCC NO: 65030; the deposit date is August 20, 2024, and it is deposited in Guangdong Provincial Microbiological Culture Collection Center, and the deposit address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou; Enterococcus faecium (GDMCC 65031): Classification and naming Enterococcus faecium , the deposit number is GDMCCNO: 65031; the deposit date is August 20, 2024, and it is deposited in Guangdong Provincial Microbiological Culture Collection Center, and the deposit address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou; Bacillus subtilis (GDMCC 65032): Classification and naming Bacillus subtilis , the deposit number is GDMCCNO: 65032; the deposit date is August 20, 2024, and it is deposited in Guangdong Provincial Microbiological Culture Collection Center, and the deposit address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0021] Example The bacterial liquid is inoculated into a sterilized solid culture medium, and the fermentation temperature is controlled at 30-35°C under oxygen isolation. The fermentation is carried out for 10 days. After the fermentation is completed, it is directly filled and stored at low temperature, and added to the chicken diet when used. The solid culture medium is made of 100 parts by weight of a traditional Chinese medicine preparation (crushed into 100 meshes), 5 parts by weight of glucose, 5 parts by weight of rice bran, 5 parts by weight of peptone, and 0.5 parts by weight of an inorganic salt, and water is added to a moisture content of 50wt%. The inorganic salt is sodium chloride and dipotassium hydrogen phosphate in a weight ratio of 1:1.

[0022] The composition of Chinese medicine preparations can be referred to in Table 1 below; Table 1 Formula of Chinese medicine preparations (unit: weight ratio) Formula 1 Formula 2 Formula 3 Codonopsis pilosula 20 10 10 Astragalus 10 15 10 Atractylodes macrocephala 10 15 10 Poria 10 10 15 Licorice 10 10 10 Tangerine peel 10 10 10 Magnolia officinalis 10 10 15 Dried ginger 10 10 10 hawthorn 10 10 10

[0023] 2 kg of bacterial liquid was added to every 100 kg of solid culture medium; the activation method of the above strains was as follows: each strain that grew well after activation was selected and inoculated into the malt juice liquid culture medium, wherein the culture temperature of Enterococcus faecium was 28-30°C and the culture condition was about 8 hours; the culture temperature of Candida tropicalis and Bacillus subtilis was 35-37°C and the culture time was about 12 hours; after measurement, the number of viable bacteria in the three bacterial liquids was 8*10 8 CFU / ml or so.

[0024] Finally, biological agents 1 to 5 were obtained, wherein: The fermentation strain of biological preparation 1 is Enterococcus faecium (GDMCC 65031), and the formula of the traditional Chinese medicine preparation is formulation 1; The fermentation strain of biological preparation 2 is Enterococcus faecium (GDMCC 65031), and the formula of the traditional Chinese medicine preparation is formulation 2; The fermentation strain of biological preparation 3 is Enterococcus faecium (GDMCC 65031), and the formula of the traditional Chinese medicine preparation is formulation 3; The fermentation strain of biological preparation 4 is Candida tropicalis (GDMCC 65030), and the formula of the traditional Chinese medicine preparation is formulation 1; The fermentation strain of biological preparation 5 is Bacillus subtilis (GDMCC 65032), and the formula of the traditional Chinese medicine preparation is formulation 1; In addition, a biological preparation 6 is provided, wherein the main body of the biological preparation is the composition 1, and 1 mg of Enterococcus faecium (GDMCC 65031) powder is additionally added to every 100 g of the composition 1.

[0025] Breeding experiment A chicken farm in Yulin, Guangxi, selected 16,000 Fengxiang Tugongyao chickens aged 34 days and randomly divided them into 8 groups. The control group was fed with a basic diet for 30 days. The proportion of biological agents added to the feed was 0.2wt%. 200 chickens were randomly grabbed and weighed in each group, and 20 were slaughtered.

[0026] The experimental groups are: Farming group 1: biological agent 1 was added to the chicken diet; Farming group 2: biological agent 2 was added to the chicken diet; Farming group 3: biological agent 3 was added to the chicken diet; Farming group 4: biological agent 4 was added to the chicken diet; Farming group 5: biological agent 5 was added to the chicken diet; Farming group 6: chickens were fed with biological agent 6; Farming group 7: Formula 1 was added to the chicken diet; Blank group: fed only with diet.

[0027] The experimental results are shown in Table 2 below; Table 2 Growth performance table Breeding Group 1 Breeding Group 2 Breeding Group 3 Breeding Group 4 Farming Group 5 Breeding Group 6 Farming Group 7 Blank group Initial weight (g) 418.13 423.19 415.08 420.52 425.74 422.33 425.25 426.16 Final weight (g) 2998.21 2919.54 2932.27 2857.31 2895.82 2902.71 2853.07 2736.11 Average daily weight gain (g) 86.00 83.21 83.91 81.23 82.34 82.68 80.93 77.00 Daily feed intake (g / day) 105.01 102.63 102.51 103.58 101.68 102.73 103.63 100.72 Feed to meat ratio 1.22 1.23 1.22 1.28 1.23 1.24 1.28 1.31 Incidence of chicken glandular gastritis (%) 15 15 20 30 30 25 30 45

[0028] Related cases of chicken glandular gastritis can be found in Figures 1 to 3 ; Figure 1 This is a photo of a healthy chicken's gizzard, which is smooth and has no ulcers; Figure 2 This is a photo of a gizzard ulcer caused by glandular gastritis in chickens; Figure 3 This is a photo of a gizzard ulcer with bleeding caused by glandular gastritis in a chicken.

[0029] Result analysis: 1. It can be seen from breeding groups 1 to 3 that there is no substantial difference in feed-to-meat ratio after different Chinese medicine formulas are fermented using the same fermentation method, which is mainly reflected in the difference in the incidence of myogenic gastritis in chickens; A comparison of breeding groups 1 to 3 with breeding groups 4 and 5 showed that fermentation with Enterococcus faecium had significant advantages in controlling the incidence of myogenic gastritis in chickens compared with fermentation with Candida tropicalis and Bacillus subtilis.

[0030] 2. By comparing the breeding groups 1 and 6, it can be seen that the unfermented Changweikang I and the probiotic Enterococcus faecium added to the feed have a certain effect of inhibiting chicken glandular gastritis. Combined with the breeding group 7, it can be seen that when the Enterococcus faecium obtained by the present invention is used as a probiotic, it can improve the feed-to-meat ratio and inhibit the occurrence of chicken glandular gastritis.

[0031] 3. Through the comparison of breeding group 4, breeding group 5 and blank group, it can be concluded that the use of our company's Changweikang I for fermentation can effectively improve the medicinal effect of Changweikang I.

[0032] From the above analysis, it can be seen that our company's Changweikang I is more easily absorbed by the chicken body after fermentation, which can improve the breeding efficiency and the effect of controlling chicken glandular gastritis; when the faecal Enterococcus screened by our company is used for fermentation or used in compound form in the form of probiotics, it can further improve the breeding efficiency and the effect of controlling chicken glandular gastritis.

[0033] The present invention illustrates the process method of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned process steps, and it does not mean that the present invention must rely on the above-mentioned process steps to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of the raw materials selected by the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A method for preparing a biological preparation for treating chicken glandular gastritis, characterized in that: The biological preparation is obtained by anaerobic fermentation of a traditional Chinese medicine preparation with Enterococcus faecium; the traditional Chinese medicine preparation is composed of the following components: Codonopsis pilosula 10-20 parts by weight; 5-15 parts by weight of Astragalus; Atractylodes macrocephala 5-15 parts by weight; Poria 5-15 parts by weight; 5-15 parts by weight of liquorice; Tangerine peel 5-15 parts by weight; 5-15 parts by weight of Magnolia officinalis; 5-15 parts by weight of dried ginger; Hawthorn 5-15 parts by weight; The classification of Enterococcus faecium is named Enterococcus faecium , the deposit number is GDMCC NO: 65031; the deposit date is August 20, 2024, and it is deposited in Guangdong Provincial Microbiological Culture Collection Center, and the deposit address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

2. The preparation method according to claim 1, characterized in that: The method is specifically as follows: Step 1: Activate Enterococcus faecium to obtain a viable count of (1~10)*10 8 CFU / ml of bacterial solution; Step 2: Inoculate the bacterial liquid into the culture medium containing the traditional Chinese medicine preparation, and in the absence of oxygen, control the fermentation temperature at 30-35°C for 10-15 days to obtain the biological preparation; The culture medium is a solid culture medium, which is composed of a crushed traditional Chinese medicine preparation, a carbon source, a nitrogen source, and an inorganic salt; and the water content of the solid culture medium is controlled at 40-60wt%.

3. The preparation method according to claim 2, characterized in that: The solid culture medium comprises 100 parts by weight of a traditional Chinese medicine preparation, 5 to 10 parts by weight of a carbon source, 3 to 5 parts by weight of a nitrogen source, and 0.5 to 1 part by weight of an inorganic salt.

4. The preparation method according to claim 3, characterized in that: The carbon source is one or more of bran, rice bran, glucose, and molasses; the nitrogen source is peptone; and the inorganic salt is one or more of K2HPO4, KH2PO4, MgSO4, NaCl, KCl, and FeSO4.

5. The preparation method according to claim 2, characterized in that: The weight ratio of the bacterial liquid to the culture medium is 1-5:

100.

6. The preparation method according to claim 2, characterized in that: The activation method in step 1 is: inoculating Enterococcus faecium into a malt juice liquid culture medium, and culturing on a shaker at 28-30° C. for 6-10 hours.

7. A biological preparation for treating chicken proventriculus, characterized in that: The method is prepared by any one of claims 1 to 6.

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