Feed additive for preventing and treating chicken infectious anemia and preparation method thereof

Abstract

The invention discloses a feed additive for preventing and treating chicken infectious anemia and a preparation method of the feed additive. The feed additive is prepared by concentrating fermentation liquor obtained by anaerobic culture of alpha-hemolytic streptococcus T33 # strains and preparing concentrated liquor serving as an active raw material. The fermentation liquor contains alpha-mannan, alpha-mannan oligosaccharide and alkaline polypeptide, wherein the content of the mannan reaches 70 to 120 mg / 100 mL. The traditional Chinese medicine composition directly acts on bone marrow to correct lymphatic atrophy, immunosuppression, anemia and other clinical symptoms caused by damage of infectious anemia virus to bone marrow, stimulates bone marrow hematopoietic stem cell proliferation in a targeted manner, quickly relieves immunosuppression caused by infectious anemia, and has the treatment effects of treating both symptoms and root causes and being accurate and efficient.

Description

Technical Field

[0001] The invention belongs to the technical field of feed additives and relates to a feed additive for preventing and treating chicken infectious anemia and a preparation method thereof. Background Art

[0002] Chicken infectious anemia (CIAV) is a disease caused by the Chicken Infectious Anemia Virus (CIAV). Also known as chicken anemia factor disease, blue wing disease, hemorrhagic syndrome, or anemic dermatitis syndrome, CIAV is an immunosuppressive disease that primarily affects chicks. Its main symptoms are aplastic anemia and systemic lymphoid tissue atrophy. It severely impacts poultry growth and egg production, posing a significant threat to the poultry industry.

[0003] Chickens are the sole reservoir for the infectious anemia virus. Chickens of all ages can be infected, with natural infection most commonly occurring in chicks aged 2 to 4 weeks. Symptoms typically develop 10 days after infection, with peak infection rates reaching 14 to 16 days. Chicks of all ages and breeds can be infected, with broilers being more susceptible than laying hens. The case fatality rate generally ranges from 5% to 60%.

[0004] Sick chickens appear depressed, weak, and slow in movement. Their beaks, wattles, facial skin, and visible mucous membranes are pale. They grow poorly, lose weight, and may also have diarrhea at the time of death. Sick chickens are anemic and emaciated, with pale muscles and internal organs and anemia. The liver and kidneys are enlarged, faded, or pale yellow, with thin blood and prolonged clotting time. The characteristic lesion at autopsy is bone marrow atrophy. The femoral bone marrow is fatty, pale yellow, or pink, and in some cases, dark red. The most common lesion is thymic atrophy, which is dark reddish-brown and may lead to complete degeneration. The bursa of Fabricius is not very atrophic and appears translucent. The glandular stomach mucosa sometimes bleeds, as do subcutaneous and muscular bleeding. Secondary bacterial infection may cause gangrenous dermatitis and mottled liver enlargement.

[0005] Infectious anemia can be transmitted both vertically and horizontally. Vertical transmission is considered the most important route of transmission, while horizontal transmission can cause infection through the oral, digestive, and respiratory tracts. The immunosuppression caused by chicken anemia virus increases the susceptibility of chickens to pathogens such as viruses, bacteria, fungi, mycoplasmas, and parasites, leading to increased losses following concurrent infection. The combined effects of exogenous and endogenous toxins exacerbate immunosuppression and increase mortality.

[0006] Infectious anemia is highly contagious and can occur year-round. It is a precursor to various disease outbreaks or multiple infections, often resulting in large numbers of chicken deaths, stunted growth, high culling rates, and low feed conversion rates among surviving chickens, bringing significant losses to my country's poultry industry.

[0007] Currently, the most commonly used treatments for infectious anemia are antiviral Chinese and Western medicines, supplemented by nutritional supplements. Clinically, administering Western medicine alone or in combination to control secondary infections after the onset of the disease can partially reduce mortality, but the treatment course is long, the efficacy is uncertain, and it falls short of therapeutic expectations. Western antiviral drugs are clinically banned. Antimicrobial drugs can only prevent secondary bacterial infections and are ineffective against viruses. They are also prohibited during the egg-laying period. Their clinical use and effectiveness are significantly restricted by the effects of drug residues and withdrawal periods before slaughter. Traditional Chinese medicines exhibit inconsistent efficacy due to differences in raw material quality, formulation processes, active ingredient content, and absorption and metabolic rates, making significant therapeutic effects difficult to achieve. Summary of the Invention

[0008] The technical problem solved by the present invention is to provide a feed additive for preventing and treating infectious anemia in chickens and a preparation method thereof. By regulating the chicken's own immunity, resisting viral infection, and reducing the body's susceptibility, the feed additive can promote the rapid recovery of sick chickens and reduce the mortality rate.

[0009] The present invention is achieved through the following technical solutions:

[0010] A feed additive for preventing and treating infectious anemia in chickens is prepared by concentrating the fermentation liquid obtained by anaerobic culture of α-hemolytic Streptococcus T33# strain (CGMCC No. 1082) and using the concentrated liquid as an active raw material.

[0011] The fermentation broth contains α-mannan, α-manno-oligosaccharide and basic polypeptide, wherein the content of mannan reaches 70-120 mg / 100 mL.

[0012] Furthermore, during the anaerobic culture, the culture medium used includes, by mass percentage, 5-15% glucose, 3-8% peptone, 3-7% sodium chloride, and the remainder is sterile water; pH 6.5-7.2;

[0013] The inoculation amount is 0.2-1%, and the culture is statically cultured at 30-37°C for 65-70 hours.

[0014] A method for preparing a feed additive for preventing and treating chicken infectious anemia comprises the following steps:

[0015] 1) α-hemolytic Streptococcus T33# strain (CGMCC No. 1082) was used as the fermentation strain;

[0016] 2) Inoculate the lyophilized strain into a test tube liquid culture medium and incubate at 28-35°C for 12-36 hours. Then inoculate the strain into a fresh broth medium test tube. Select strains with vigorous growth, strong vitality, and morphological characteristics that meet the requirements to prepare the test tube liquid culture.

[0017] 3) Aseptically inoculate the liquid culture in the test tube into a shake flask medium at a 0.2-1% inoculum volume and incubate at 37°C for 24 hours.

[0018] 4) Inoculate the shake flask medium into the fermentation medium under sterile conditions at a 0.2-1% inoculum level and incubate at 30-37°C for 65-70 hours. Incubate under a microscopic examination. When the reducing sugars and amino nitrogen are almost depleted, inactivate the cells in a water bath at 80-85°C for 30 minutes.

[0019] In terms of mass percentage, the fermentation medium comprises 5-15% glucose, 3-8% peptone, 3-7% sodium chloride, and the remainder is sterile water; pH 6.5-7.2;

[0020] 5) The inactivated fermentation broth was purified by nanofiltration with a maximum pressure of 1.1-1.4 MPa and a minimum pressure of 0.1-0.15 MPa;

[0021] 6) The filtrate is concentrated by double-effect vacuum to a concentration ratio of 20-60:1 to obtain the active raw material;

[0022] 7) The active raw materials are prepared into feed additives for preventing and treating chicken infectious anemia.

[0023] Furthermore, in parts by mass, the preparation is:

[0024] 1-10 parts of active raw materials are diluted with 90-99 parts of purified water, sterilized, and packaged to prepare a liquid additive;

[0025] Alternatively, 1-10 parts of the active raw material are spray-dried, and then 90-99 parts of rice husk as a carrier are added and mixed thoroughly to prepare a solid feed additive;

[0026] Alternatively, 1-10 parts of the active raw material are spray-dried, and then 90-99 parts of an inorganic salt as a carrier are added and fully mixed to prepare a solid feed additive.

[0027] The prepared feed additive for preventing and treating chicken infectious anemia is used in the preparation of feed additives or preparations for preventing and treating chicken infectious anemia.

[0028] Compared with the prior art, the present invention has the following beneficial technical effects:

[0029] The feed additive for preventing and treating infectious anemia in chickens provided by the present invention is obtained by deep culturing the α-hemolytic Streptococcus T33# strain (CGMCC No. 1082) obtained by space-borne mutagenesis to obtain an immunomodulatory substance with strong specificity and extremely high biological activity: a complex of α-mannan, α-mannose oligosaccharide, and basic polypeptide.

[0030] The feed additive for preventing and treating infectious anemia in chickens provided by the present invention can increase the weight of immune organs, increase the number and activity of immune cells, increase the number and activity of immune substances, comprehensively improve the body's immunity, resist infection of infectious anemia in chickens, and can prevent and treat both the symptoms and the root causes: accurately targeting pathogens, quickly controlling the spread of the disease, greatly reducing mortality losses, with stable and efficient efficacy, no toxic side effects, no residue, easy to use, high cost-effectiveness, comprehensively improving the prevention and control effect of infectious anemia in chickens, protecting the safety and health of the chicken flock, and ensuring a stable supply of meat and egg production.

[0031] Unlike existing technical solutions for controlling chicken infectious anemia with Western medicine or a combination of Chinese and Western medicine, the present invention aims to regulate the chicken's own immunity, resist viral infection, reduce the body's susceptibility, reduce mortality, and promote rapid recovery. The additives provided have clear targeting, stable effects, and rapid control. There is no drug residue and no drug resistance, which can overcome the defects and efficacy constraints of existing solutions.

[0032] The feed additive of the present invention has a strong immunomodulatory effect, which can enhance immunity in chickens after administration. The present invention can control the onset of clinical symptoms of chicken infectious anemia and reduce high mortality to normal levels within about 4-7 days, and has a precise, stable, and rapid therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1-1 to Figure 1-2 Comparison of the effects of the feed additive of the present invention on chicken intestinal mucosa;

[0034] Figure 2-1 to Figure 2-2 This is a comparison of the immune effects of the feed additive of the present invention on chicken thymus;

[0035] Figure 3-1 to Figure 3-2 This is a comparison of the effects of the feed additive of the present invention on chicken bursa of Fabricius;

[0036] Figure 4-1 to Figure 4-2 This is a comparison of the effects of the feed additive of the present invention on chicken spleen;

[0037] Figure 5 The following is a comparison of the therapeutic effects of the feed additive of the present invention;

[0038] Figure 6 This is the appearance of a chicken infected with chicken infectious anemia. DETAILED DESCRIPTION

[0039] The present invention is further described in detail below with reference to the embodiments, which are intended to explain the present invention rather than to limit it.

[0040] The present invention provides a feed additive for preventing and treating infectious anemia in chickens, which is obtained by concentrating the fermentation liquid obtained by anaerobic culture of α-hemolytic Streptococcus T33# strain (CGMCC No.1082), and preparing the feed additive using the concentrated liquid as the active raw material;

[0041] The fermentation broth contains α-mannan, α-manno-oligosaccharide and basic polypeptide, wherein the content of mannan reaches 70-120 mg / 100 mL.

[0042] During the anaerobic culture, the culture medium used includes, by mass percentage, 5-15% glucose, 3-8% peptone, 3-7% sodium chloride, and the remainder is sterile water; pH 6.5-7.2;

[0043] The inoculation amount is 0.2-1%, and the culture is statically cultured at 30-37°C for 65-70 hours.

[0044] The invention uses α-hemolytic Streptococcus T33# strain (CGMCC No. 1082, see CN1635110A) for deep anaerobic culture, purifies and concentrates its fermentation culture fluid, and obtains highly active metabolites including α-mannan, α-mannose oligosaccharide and basic polypeptide.

[0045] The preparation method of the feed additive for preventing and treating chicken infectious anemia provided by the present invention comprises the following steps:

[0046] 1. Bacterial strain: α-hemolytic Streptococcus T33# strain (CGMCC No. 1082).

[0047] 2. Inoculate the strains in the freeze-dried tube into a test tube liquid culture medium (broth culture medium), culture at 28-35°C for 12-36 hours, and then inoculate into a fresh broth culture medium test tube after passing the sterility test and morphological identification. Repeat the above conditions twice, select strains with vigorous growth, strong vitality, and morphological characteristics that meet the requirements, and store them in the refrigerator.

[0048] 3. Inoculate the excellent test tube liquid bacteria into the shake flask culture medium (broth culture medium) under sterile conditions at an inoculation rate of 0.2-1%, incubate at 37°C for 24 hours, and examine under a microscope to ensure good growth and no foreign bacteria.

[0049] 4. Aseptically inoculate the fermentation medium at a 0.2-1% inoculum volume and incubate at 30-37°C for 65-70 hours. Microscopically, when the staining begins to fade, the coiled ball chains begin to partially break, and the reducing sugars and amino nitrogen are nearly depleted, inactivate the bacteria in a water bath at 80°C for 30 minutes.

[0050] The fermentation medium is the most suitable medium for the strain to produce mannan, and comprises, by mass percentage, 5-15% glucose, 3-8% peptone, 3-7% sodium chloride, and the rest is sterile water; the pH value is 6.5-7.2.

[0051] With an inoculation amount of 0.2-1% and static culture at 37°C for 65-70 hours, the content of mannan in the fermentation broth can reach 70-120 mg / 100 mL, thereby increasing the yield of the mannan product and achieving the purpose of optimizing the culture medium.

[0052] 5. The inactivated fermentation liquid is filtered and purified by nanofiltration membrane, with the highest pressure of 1.1-1.4Mpa and the lowest pressure of 0.1-0.15Mpa, so that the transmittance and content meet the requirements.

[0053] 6. The filtrate is concentrated by double-effect vacuum to a concentration ratio of 20-60:1; a product with a higher concentration of active product is obtained, which is called active raw material.

[0054] 7. Prepare the following preparations in parts by mass:

[0055] [1] 1-10 parts of active ingredients are diluted with 90-99 parts of purified water, sterilized, and packaged to make a liquid additive for chicken drinking water.

[0056] [2] Alternatively, 1-10 parts of the active ingredient are spray-dried and then added with 90-99 parts of rice husk as a carrier. The mixture is thoroughly mixed to prepare a solid feed additive for use in chicken feed.

[0057] [3] Alternatively, 1-10 parts of the active raw material are spray-dried and then added with 90-99 parts of an inorganic salt (such as anhydrous sodium sulfate) as a carrier. The mixture is thoroughly mixed to prepare a solid feed additive for use in chicken feed or drinking water.

[0058] Solid feed additive (carrier rice husk / inorganic salt): 100g / 1000kg body weight, used continuously for 5-7 days.

[0059] Liquid feed additives were administered in drinking water at a dose of 200 ml / 1000 kg body weight for 5-7 consecutive days.

[0060] The feed additive provided by the present invention is a highly active immunomodulator that directly acts on the bone marrow, corrects the damage to the bone marrow caused by the infectious anemia virus, and causes clinical symptoms such as lymph atrophy, immunosuppression, and anemia. It also stimulates the proliferation of bone marrow hematopoietic stem cells in a targeted manner, and quickly relieves the immunosuppression caused by infectious anemia, achieving a precise and efficient therapeutic effect of treating both the symptoms and the root cause.

[0061] The invention also has powerful mucosal protection and intestinal conditioning effects, can protect the integrity of the mucosa, enhance its barrier function, promote the chickens to recover health as soon as possible, and reduce secondary infections or complications.

[0062] like Figure 1-1 、 1-2 As shown in the comparison, Figure 1-2 The control model (not using the product of the present invention) has a rough and cracked intestinal mucosa, a thin mucus layer, upright columnar intestinal villi, no differentiated branches or crossings, few intestinal glands in the lamina propria, a small number of immune cells, and a thin inner circular muscle layer. Figure 1-1 After using the feed additive of the present invention, the intestinal mucosa is intact and smooth, the mucus layer is thick, the intestinal villi are fully developed, branched and crossed, the intestinal glands in the propria are well developed, the number of immune cells is large, and the inner circular muscle layer is thick.

[0063] Comparative studies on chickens revealed the immunomodulatory mechanism of the feed additive of the present invention, and its immunomodulatory effects include:

[0064] Central immune regulation - the effect of the thymus:

[0065] like Figure 2-1 、 Figure 2-2 As shown in the comparison, Figure 2-2 For the control group (not using the product of the present invention, HE staining): the chicken thymus structure is not clear enough, the cortex (blue) and medulla (red) have loose structures and unclear boundaries, the lymphocyte density is low, the thymus structure is poorly intact, and the development is not sufficient.

[0066] After using the product of the present invention Figure 2-1 As shown, in the experimental group (HE staining): the chicken thymus has a clear structure, the cortex (blue) and medulla (red) are densely structured, with clear boundaries, a high density of lymphocytes, good thymus structural integrity, and full development.

[0067] Central immune regulation - the effect of the bursa of Fabricius:

[0068] like Figure 3-1 、 Figure 3-2 As shown in the comparison, Figure 3-2 Control group (HE staining): The chicken bursa of Fabricius has narrow and thin folds, loose arrangement, loose structure of lymphoid follicles (polygonal), unclear boundary between cortex and medulla, low density of lymphocytes, poor structural integrity of bursa of Fabricius, and insufficient development. Figure 3-1 For the test group using the present invention (HE staining): the chicken bursa of Fabricius has thick and dense folds, densely arranged lymphoid follicles (polygonal) structure, clear boundaries between the cortex and medulla, high density of lymphocytes, good structural integrity of the bursa of Fabricius, and sufficient development.

[0069] Peripheral immune regulation - effects on the spleen:

[0070] like Figure 4-1 、 Figure 4-2 As shown in the comparison, Figure 4-2 For the control group (HE staining): the structure of the chicken spleen is not clear enough, the structure is loose, the boundary between the red pulp (light color) and the white pulp (dark color) is not clear, the density of lymphocytes is low, and the spleen structure integrity is poor. Figure 4-1 For the experimental group (HE staining): the chicken spleen has a clear and dense structure, a clear boundary between the red pulp (light color) and the white pulp (dark color), a high density of lymphocytes, and good spleen structural integrity.

[0071] In addition, the feed additive of the present invention has significant effects on protecting the liver and kidneys, has a strong detoxification effect, can reduce the harm of internal and external toxins to chickens, reduce the incidence of diseases, and reduce losses.

[0072] The feed additive of the present invention also has a strong anti-stress effect, can reduce the harm of various stresses to chickens, and enable the chickens to grow and produce healthily.

[0073] Specific prevention and control examples are given below.

[0074] 1. A breeding group in South China in April 2020

[0075] Infectious anemia problems began to appear in a certain branch farm in April. The chickens showed symptoms such as white combs and feet, listlessness, shrunken heads, drooping wings, and yellow bone marrow during clinical dissection.

[0076] The feed additive of the present invention was used for treatment, and the treated group and the untreated group were compared in parallel. The treatment effect was shown in FIG. Figure 5 , where both treatment groups 1 and 2 were administered with the feed additive of the present invention, with the untreated group serving as the control. After 8 consecutive days of administration, compared to the untreated group, when the mortality rate in the untreated group was significantly increased, the control effect in the treatment group was significant; and after 33 days, the present invention still had a control effect.

[0077] 2. April 2024, a group in North China

[0078] Broiler farmers’ chickens are sick (the appearance of sick chickens is as follows Figure 6 The liquid preparation of the present invention was used as a single treatment, with concentrated drinking water every day for 8 consecutive days, and the whole drinking time was 2 hours.

[0079] Drink water according to weight ratio.

[0080] Treatment results: The daily mortality rates at the onset of disease were 0.718% and 0.471% (two groups of chicken sheds), and after treatment, the daily mortality rates were 0.102% and 0.059% respectively;

[0081] After treatment, the daily mortality rate was reduced by 7 and 8 times respectively. Chicken infectious anemia was effectively controlled, the symptoms gradually disappeared, and the chickens' spirits, feeding and growth recovered.

[0082] 3. June 2024: A farmer

[0083] A flock of 120,000 broiler chickens fell ill and was diagnosed as chicken infectious anemia by a laboratory. The chickens were treated with the product of the present invention by a centralized drinking water method, and the whole chickens were drunk within 2 hours for 5 consecutive days.

[0084] The daily mortality rates before and after treatment were 0.123% and 0.0175% respectively. After treatment, the daily mortality rate dropped by 7 times. The chickens recovered, and their feeding and growth returned to normal. The market performance was: average weight 2.61 kg / bird, and the European index was 412.

[0085] In summary, the feed additive for preventing and treating chicken infectious anemia provided by the present invention has good prevention and treatment effects.

[0086] The present invention also proposes the use of the prepared feed additive for preventing and treating chicken infectious anemia in the preparation of feed additives or preparations

[0087] The above embodiments are preferred examples for implementing the present invention, and the present invention is not limited to the above embodiments. Any non-essential additions or replacements made by those skilled in the art based on the technical features of the present invention fall within the scope of protection of the present invention.

Claims

1. A feed additive for preventing and treating chicken infectious anemia, characterized in that: This is a feed additive obtained by concentrating the fermentation liquid obtained by anaerobic culture of α-hemolytic Streptococcus T33# strain (CGMCC No.1082), and using the concentrated liquid as the active raw material; The fermentation broth contains α-mannan, α-manno-oligosaccharide and basic polypeptide, wherein the content of mannan reaches 70-120 mg / 100 mL.

2. The feed additive for preventing and treating chicken infectious anemia according to claim 1, characterized in that: During the anaerobic culture, the culture medium used includes, by mass percentage, 5-15% glucose, 3-8% peptone, 3-7% sodium chloride, and the remainder is sterile water; pH 6.5-7.2; The inoculation amount is 0.2-1%, and the culture is statically cultured at 30-37°C for 65-70 hours.

3. The feed additive for preventing and treating chicken infectious anemia according to claim 1, wherein: In parts by mass, the preparation is: 1-10 parts of active raw materials are diluted with 90-99 parts of purified water, sterilized, and packaged to prepare a liquid additive; Alternatively, 1-10 parts of the active raw material are spray-dried, and then 90-99 parts of rice husk / inorganic salt as a carrier are added and fully mixed to prepare a solid feed additive.

4. A method for preparing a feed additive for preventing and treating chicken infectious anemia, characterized in that: The following operations are included: 1) α-hemolytic Streptococcus T33# strain (CGMCC No. 1082) was used as the fermentation strain; 2) Inoculate the lyophilized strain into a test tube liquid culture medium and incubate at 28-35°C for 12-36 hours. Then inoculate the strain into a fresh broth medium test tube. Select strains with vigorous growth, strong vitality, and morphological characteristics that meet the requirements to prepare the test tube liquid culture. 3) Aseptically inoculate the liquid culture in the test tube into a shake flask medium at a 0.2-1% inoculum volume and incubate at 37°C for 24 hours. 4) Inoculate the shake flask medium into the fermentation medium under sterile conditions at a 0.2-1% inoculum level and incubate at 30-37°C for 65-70 hours. Incubate under a microscopic examination. When the reducing sugars and amino nitrogen are almost depleted, inactivate the cells in a water bath at 80-85°C for 30 minutes. In terms of mass percentage, the fermentation medium comprises 5-15% glucose, 3-8% peptone, 3-7% sodium chloride, and the remainder is sterile water; pH 6.5-7.2; 5) The inactivated fermentation broth was purified by nanofiltration with a maximum pressure of 1.1-1.4 MPa and a minimum pressure of 0.1-0.15 MPa; 6) The filtrate is concentrated by double-effect vacuum to a concentration ratio of 20-60:1 to obtain the active raw material; 7) The active raw materials are prepared into feed additives for preventing and treating chicken infectious anemia.

5. The method for preparing the feed additive for preventing and treating chicken infectious anemia according to claim 4, wherein: The content of mannan in the fermentation broth reaches 70-120 mg / 100 mL.

6. The method for preparing the feed additive for preventing and treating chicken infectious anemia according to claim 4, wherein: In parts by mass, the preparation is: 1-10 parts of active raw materials are diluted with 90-99 parts of purified water, sterilized, and packaged to prepare a liquid additive; Alternatively, 1-10 parts of the active raw material are spray-dried, and then 90-99 parts of rice husk as a carrier are added and mixed thoroughly to prepare a solid feed additive; Alternatively, 1-10 parts of the active raw material are spray-dried, and then 90-99 parts of an inorganic salt as a carrier are added and fully mixed to prepare a solid feed additive.

7. Use of the feed additive for preventing and treating chicken infectious anemia prepared according to claim 4 in the preparation of a feed additive or preparation for preventing and treating chicken infectious anemia.

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