A composite microbial feed additive and preparation method thereof

Abstract

The present application relates to the technical field of feed additives, and specifically discloses a composite microbial feed additive and a preparation method thereof. The composite microbial feed additive comprises the following components in parts by weight: 10-30 parts of sodium butyrate; 30-50 parts of silicon dioxide; 40-60 parts of bran; 60-80 parts of modified potato starch; 7-15 parts of Clostridium butyricum; 4-9 parts of Lactobacillus plantarum; and 2-6 parts of Bacillus licheniformis. The composite microbial feed additive provided in the present application can effectively increase the daily feed intake and daily weight gain of broilers, reduce the feed-to-weight ratio, and reduce the mortality rate of broilers.

Description

Technical Field

[0001] The present application relates to the technical field of feed additives, and more specifically, to a composite microbial feed additive and a preparation method thereof. Background Art

[0002] The intestine is the largest digestive organ of poultry and is the place for digestion and absorption of nutrients. The surface of the intestinal mucosa is covered with intestinal villi. The food entering the intestine is broken down into glucose and amino acids under the action of various digestive juices. After the food is digested and absorbed, the remaining material forms feces and is excreted from the body. Once the normal function of the intestine is affected, it will cause food digestion and nutrient absorption disorders, thereby affecting the growth of broilers. Studies have shown that the combination or interaction of different viruses, different bacteria and other infectious or non-infectious pathogens is sufficient to induce intestinal diseases of varying degrees. Therefore, broiler intestinal diseases have always plagued the development of the broiler industry.

[0003] At present, the method of adding feed antibiotics is usually used to prevent such diseases, but the use of antibiotics has led to many problems, such as drug residues, drug resistance, superinfection and reduced immunity, and its application has been greatly limited. Therefore, it is necessary to develop a new solution to replace antibiotics.

[0004] Clostridium butyrium is a Gram-positive anaerobic bacillus belonging to the genus Clostridium, which exists in soil, cheese, animal feces, etc. In addition to being widely used in medicine, food and feed, Clostridium butyrium is also used as a microbial fertilizer in Japan, which can prevent diseases and promote the growth of crops.

[0005] Clostridium butyricum can colonize in the intestines of animals and produce beneficial products mainly composed of short-chain fatty acids such as butyric acid and lactic acid. It can promote the proliferation and development of beneficial bacteria in the intestines of animals, inhibit the growth and reproduction of spoilage bacteria and harmful bacteria (cholera, dysentery, salmonella, pathogenic Escherichia coli), correct intestinal flora disorders, reduce and lower the production of enterotoxins, and has the effects of improving animal growth performance, improving feed digestibility, and regulating the balance of animal intestinal microecology. It has been approved by the European Union in 2003 as a feed additive for broilers and weaned piglets, and has been gradually promoted as a probiotic feed additive.

[0006] Existing studies have found that adding xylooligosaccharides and Clostridium butyricum as additives to feed can improve feed utilization. However, in actual applications, it is found that although the simple combination of xylooligosaccharides and Clostridium butyricum can achieve certain effects, the effects achieved are very limited, and a large number of other effective ingredients need to be added to improve the feeding effect. In addition, the number of live bacteria and spore rate of current Clostridium butyricum products are low, and the effect is limited. In the production of xylooligosaccharides, acid-base pretreatment and complex multiple enzymatic hydrolysis processes are required, the process is complicated, and the cost of feed is high, which makes it difficult to be widely used on a large scale in the feed industry.

[0007] Based on the above, it is necessary to provide a new feed additive to replace antibiotics to prevent poultry intestinal diseases. Summary of the invention

[0008] The present application provides a composite microbial feed additive and a preparation method thereof. The composite microbial feed additive can effectively increase the daily feed intake and daily weight gain of broilers, reduce the feed-to-weight ratio, and reduce the mortality rate of broilers.

[0009] In the first aspect, the present application provides a composite microbial feed additive, which adopts the following technical solution:

[0010] A composite microbial feed additive comprises the following components in parts by weight: 10-30 parts of sodium butyrate; 30-50 parts of silicon dioxide; 40-60 parts of bran; 60-80 parts of modified potato starch; 7-15 parts of Clostridium butyricum; 4-9 parts of Lactobacillus plantarum; and 2-6 parts of Bacillus licheniformis.

[0011] The present application provides a composite microbial feed additive of Clostridium butyricum, Lactobacillus plantarum and Bacillus licheniformis, which can effectively increase the daily feed intake and daily weight gain of broilers and reduce the feed-to-weight ratio. At the same time, the composite microbial feed additive provided by the present application can also significantly reduce the mortality rate of broilers.

[0012] Clostridium butyricum can promote the proliferation and development of beneficial flora in the intestines of animals, inhibit the growth and reproduction of harmful bacteria and putrefactive bacteria in the intestines, correct intestinal flora disorders, and reduce the occurrence of enterotoxins; at the same time, it can produce B vitamins, vitamin K, amylase and other substances in the intestines of animals, which have health-care effects; in addition, butyric acid, the main metabolite of Clostridium butyricum, is the main nutrient for the regeneration and repair of intestinal epithelial tissue cells; more importantly, Clostridium butyricum is an anaerobic or facultative anaerobic Bacillus, and is not affected by gastric acid, bile acid, etc.

[0013] Bacillus licheniformis is a facultative anaerobic microorganism that can produce spores. It is an excellent microbial strain widely used in medicine, food and feed for microecological preparations. After entering the intestine, Bacillus licheniformis will grow and multiply rapidly. At the same time, its growth and reproduction have a biological oxygen consumption effect, causing a low oxygen state in the ecological environment. By supporting the proliferation of physiological anaerobic bacteria, inhibiting the growth of pathogenic bacteria, correcting the disorder of intestinal flora, and restoring intestinal function.

[0014] Lactobacillus plantarum is an anaerobic or facultative anaerobic lactic acid bacterium that is widely used in the food industry as a fermentation agent and preservative. Due to its excellent fermentation characteristics and preservative properties, Lactobacillus plantarum plays an important role in food, providing food with a unique flavor and texture, while also extending the shelf life of food. Appropriate intake of Lactobacillus plantarum can improve blood lipid metabolism, lower cholesterol levels, and prevent the occurrence of cardiovascular diseases. At the same time, Lactobacillus plantarum can also inhibit intestinal inflammatory reactions and reduce the occurrence of intestinal inflammation.

[0015] In the present application, Clostridium butyricum and Lactobacillus plantarum are both anaerobic or facultative anaerobic bacillus, and the growth and reproduction of Bacillus licheniformis will create a low oxygen environment, thus creating favorable conditions for the growth and reproduction of Clostridium butyricum and Lactobacillus plantarum. Therefore, the composite use of the three can promote the growth and reproduction of the three, thereby further improving the effects of the three strains in the intestine, thereby further promoting the digestion and absorption of food by the intestine and improving the utilization rate of feed.

[0016] In addition, sodium butyrate can promote the development of intestinal mucosa, repair mucosal epithelial cells, activate lymphocytes, and reduce gastrointestinal pH. Sodium butyrate and Clostridium butyricum promote and overlap each other, which is more conducive to the decomposition of butyric acid, promote intestinal development and flora balance, reduce the occurrence of intestinal problems, and promote the control of intestinal pH in an acidic environment to ensure intestinal health.

[0017] The main function of silicon dioxide in feed additives is to prevent sodium butyrate from agglomerating and reduce the adhesion between bran, so that the distribution of sodium butyrate and bran in the mixture is more uniform. After the potato modified starch is mixed, the bran and sodium butyrate can be more evenly distributed in the small granular material or on the surface of the small granular material. At the same time, due to the presence of bran, the surface of the small granular material will be rougher, so it is easy to wrap Clostridium butyricum, Lactobacillus plantarum and Bacillus licheniformis on the surface of the small granular material particles.

[0018] The main component of bran is dietary fiber, so it can increase the roughness of the surface of small granular substances. Clostridium butyricum, Lactobacillus plantarum and Bacillus licheniformis can attach to the gaps between dietary fibers, thus increasing the surface area to which Clostridium butyricum, Lactobacillus plantarum and Bacillus licheniformis can attach, thereby more effectively providing more attachment points for the bacteria. The main components of diatomaceous earth and zeolite powder are both silicon dioxide. The surface roughness of the small granular substances formed by them is low, and the bacteria not only attach to the surface, but also may pass through potato starch into the small granular substances, which may cause the bacteria to not fully function. Therefore, the effect of the composite microbial feed additive made of diatomaceous earth and zeolite powder is far worse than that of the composite microbial feed additive made of bran.

[0019] Optionally, the weight portion of the Clostridium butyricum in the composite microbial feed additive is 10-12 parts.

[0020] In a specific embodiment, the weight proportion of Clostridium butyricum is 7 parts, 10 parts, 11 parts, 12 parts, or 13 parts.

[0021] In some specific embodiments, the weight proportion of Clostridium butyricum is 7-10 parts, 7-11 parts, 7-12 parts, 10-11 parts, 10-12 parts, 10-13 parts, 11-12 parts, 11-13 parts, 12-13 parts.

[0022] Optionally, the weight portion of the Lactobacillus plantarum in the composite microbial feed additive is 5.5-7.5 parts.

[0023] In a specific embodiment, the weight proportion of the Lactobacillus plantarum is 4 parts, 5.5 parts, 6.5 parts, 7.5 parts, or 9 parts.

[0024] In some specific embodiments, the weight proportion of the Lactobacillus plantarum is 4-5.5, 4-6.5, 4-7.5, 5.5-6.5, 5.5-7.5, 5.5-9, 6.5-7.5, 6.5-9, or 7.5-9.

[0025] Optionally, the weight portion of the Bacillus licheniformis in the composite microbial feed additive is 2.5-5.5 parts.

[0026] In a specific embodiment, the weight proportion of the Bacillus licheniformis is 2 parts, 2.5 parts, 4 parts, 5.5 parts, or 6 parts.

[0027] In some specific embodiments, the weight proportion of the Bacillus licheniformis is 2-2.5 parts, 2-4 parts, 2-5.5 parts, 2.5-4 parts, 2.5-5.5 parts, 2.5-6 parts, 4-5.5 parts, 4-6 parts, or 5.5-6 parts.

[0028] Optionally, the Clostridium butyricum is selected from any one or more of the deposit numbers CGMCC NO. 25504, CGMCC NO. 27479 and CGMCC NO. 27311.

[0029] In a specific embodiment, the deposit number of the Clostridium butyricum is CGMCC NO. 25504.

[0030] In a specific embodiment, the deposit number of the Clostridium butyricum is CGMCC NO. 27479.

[0031] In a specific embodiment, the deposit number of the Clostridium butyricum is CGMCC NO. 27311.

[0032] The related art discloses that Clostridium butyricum with a deposit number of CGMCC NO. 25504 can be used as a host for recombinant expression vectors in genetic engineering, and can also be used in the preparation of drugs for preventing or treating discomfort caused by flora disorders after eating hot pot and drinking. However, there is no application of the strain with this deposit number in composite microbial feed additives.

[0033] The related art discloses the application of Clostridium butyricum with a deposit number of CGMCC NO. 27479 in the field of biological hydrogen production, especially anaerobic dark fermentation hydrogen production, but there is no application of the strain with this deposit number in composite microbial feed additives.

[0034] In the related art, the application direction of Clostridium butyricum with the deposit number CGMCC NO. 27311 has not been explored, so the bacteria with this deposit number are also used in composite microbial feed additives.

[0035] Optionally, the plant lactobacillus is selected from any one or more of the collection numbers CGMCC NO. 10632, CGMCC NO. 24430 and CGMCC NO. 11262.

[0036] In a specific embodiment, the deposit number of the Lactobacillus plantarum is CGMCC NO. 10632.

[0037] In a specific embodiment, the deposit number of the Lactobacillus plantarum is CGMCC NO. 24430.

[0038] In a specific embodiment, the deposit number of the plant lactobacillus is CGMCC NO. 11262.

[0039] The related art discloses that the plant lactobacillus with a deposit number of CGMCC NO. 10632 has strong adhesion and cholesterol-lowering abilities, good acid and bile salt resistance, and can ferment fresh milk (or reconstituted milk) to produce a coagulated yogurt with a thick taste and a distinct and fresh aroma, providing a new fermentation bacteria source for the production of yogurt products and the development of functional yogurt. However, there is no application of the bacteria with the deposit number in composite microbial feed additives.

[0040] The related art discloses that Lactobacillus plantarum with a preservation number of CGMCC NO. 24430 has a significant inhibitory effect on Phytophthora capsici, and has a significant inhibitory effect on red anthracnose bacteria of pepper, Escherichia coli, Salmonella, Shigella and Enterobacter sakazakii. It also has a significant inhibitory effect on Phytophthora capsici after harvesting peppers. Therefore, it has a good application prospect in the preservation and freshness of post-harvest peppers. However, the application of the strain with this preservation number in composite microbial feed additives has not been seen.

[0041] The related art discloses that Lactobacillus plantarum with a deposit number of CGMCC NO. 11262 has a high antibacterial ability, especially in the determination of the ability to inhibit pathogenic Escherichia coli, and has obvious advantages over other strains. Animal experiments have found that it can effectively promote the production performance of livestock and poultry, and can be used as a feed additive and fermentation agent in livestock and poultry breeding. It is also disclosed that it and Bacillus subtilis are used as bacterial agents in fattening beef cattle breeding. However, there is no case of compounding the strain with this deposit number with other types of strains.

[0042] Optionally, the Bacillus licheniformis is selected from any one or more of the deposit numbers CGMCC NO. 20892, CGMCC NO. 18463 and CGMCC NO. 22062.

[0043] In a specific embodiment, the deposit number of the Bacillus licheniformis is CGMCC NO. 20892.

[0044] In a specific embodiment, the deposit number of the Bacillus licheniformis is CGMCC NO. 18463.

[0045] In a specific embodiment, the deposit number of the Bacillus licheniformis is CGMCC NO. 22062.

[0046] The related art discloses that the Bacillus licheniformis with a deposit number of CGMCC NO. 20892 can be used for biodegradation and detoxification of feed, and can effectively prevent and treat animal diarrhea; however, there is no evidence of the fattening effect of the strain with this deposit number in livestock and poultry farming, nor is there any evidence of the strain with this deposit number being compounded with other such strains.

[0047] The related art discloses that Bacillus licheniformis with a deposit number of CGMCC NO. 18463 can significantly improve the body color of clownfish, increase the growth rate and survival rate of clownfish, and is a green and safe aquatic feed additive. However, there is no application of the strain with this deposit number in composite microbial feed additives, nor is there any fattening effect of the strain with this deposit number in livestock and poultry breeding.

[0048] The related art discloses that Bacillus licheniformis with a deposit number of CGMCC NO. 22062 can efficiently degrade soybean antigen protein, with a degradation rate of 95.94% for soybean globulin and 92.33% for β-conglycinin, which is significantly higher than the degradation rate of antigen protein by fermentation of other strains, and the degradation effect is significant. However, there is no application of the strain with the deposit number in composite microbial feed additives.

[0049] Optionally, the composite microbial feed additive contains 350-600 million cfu / g of Clostridium butyricum.

[0050] Optionally, the composite microbial feed additive contains 150-450 million cfu / g of Lactobacillus plantarum.

[0051] Optionally, the composite microbial feed additive contains 0.5-3.0 billion cfu / g of Bacillus licheniformis.

[0052] Optionally, the weight portion of the bran is 45-55 parts.

[0053] In a second aspect, the present application provides a method for preparing a composite microbial feed additive, which specifically comprises the following steps: fully mixing the sodium butyrate, the silicon dioxide and the bran, and then spray-drying the obtained mixture and the potato modified starch to form a small granular substance, and then fully mixing the small granular substance with the Clostridium butyricum, the Lactobacillus plantarum and the Bacillus licheniformis to obtain the composite microbial feed additive.

[0054] Optionally, the particle size of the small granular material is 0.2-0.5 mm.

[0055] Optionally, the Clostridium butyricum, the Lactobacillus plantarum, and the Bacillus licheniformis are all in powder form.

[0056] In a third aspect, the present application provides a feed comprising the above-mentioned composite microbial feed additive.

[0057] In summary, this application has the following beneficial effects:

[0058] 1. The composite microbial feed additive provided by the present application can effectively increase the daily feed intake and daily weight gain of broilers, reduce the feed-to-weight ratio, and the weight of 42-day broilers is significantly different from that of the control group. At the same time, compared with the control group, the composite microbial feed additive provided by the present application can also significantly reduce the mortality rate of broilers.

[0059] 2. Compared with adding only one or two of Clostridium butyricum, Lactobacillus plantarum or Bacillus licheniformis to the feed additive, adding the above three species of bacteria at the same time in the composite microbial feed additive of the present application can effectively increase the daily feed intake and daily weight gain of broilers, reduce the feed-to-weight ratio, and significantly reduce the mortality rate of broilers.

[0060] 3. The present application screened strains of Clostridium butyricum, Lactobacillus plantarum or Bacillus licheniformis. From the screening results, it can be seen that when Clostridium butyricum with a preservation number of CGMCC NO. 27479, Lactobacillus plantarum with a preservation number of CGMCC NO. 24430 and Bacillus licheniformis with a preservation number of CGMCC NO. 18463 are selected, the daily feed intake and daily weight gain of broilers can be further increased, and the feed-to-weight ratio can be reduced.

[0061] 4. After testing, it was found that the daily feed intake, daily weight gain and feed-to-weight ratio of broilers when only three strains or bran were added were far worse than those when the three strains and bran were added at the same time. And when the bran was replaced with diatomaceous earth or zeolite powder, the daily feed intake, daily weight gain and feed-to-weight ratio of broilers were also not as good as those when the three strains and bran were added at the same time. It can be seen that in the scheme of the present application, the simultaneous addition of bran and three strains can further effectively increase the daily feed intake and daily weight gain of broilers and reduce the feed-to-weight ratio.

[0062] 5. The feed additive provided in this application can effectively promote the intestinal development of broiler chickens and improve the absorption of nutrients. DETAILED DESCRIPTION

[0063] Before describing the embodiments of the present application in detail, it should be understood that the terms used herein are only used for the purpose of describing specific embodiments. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by ordinary technicians in the field to which the terms belong.

[0064] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. Further, in the description of this application, unless otherwise specified, the meaning of "plurality" is two or more.

[0065] The endpoints and any values ​​of the ranges disclosed in this application are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0066] In the present application, the terms "comprise" or "include" are open expressions, that is, including the contents specified in the present application but not excluding other contents.

[0067] The present application provides a composite microbial feed additive, which specifically comprises the following components by weight: 10-30 parts of sodium butyrate; 30-50 parts of silicon dioxide; 40-60 parts of bran; 60-80 parts of modified potato starch; 7-15 parts of Clostridium butyricum; 4-9 parts of Lactobacillus plantarum; and 2-6 parts of Bacillus licheniformis.

[0068] Furthermore, Clostridium butyricum is selected from any one or more of the collection numbers CGMCC NO. 25504, CGMCC NO. 27479 and CGMCC NO. 27311.

[0069] Furthermore, the plant lactobacillus is selected from any one or more of the collection numbers CGMCC NO. 10632, CGMCC NO. 24430 and CGMCC NO. 11262.

[0070] Furthermore, the licheniformis Bacillus is selected from any one or more of the deposit numbers CGMCC NO. 20892, CGMCC NO. 18463 and CGMCC NO. 22062.

[0071] Furthermore, the composite microbial feed additive contains 350-600 million cfu / g of Clostridium butyricum, 150-450 million cfu / g of Lactobacillus plantarum, and 50-300 million cfu / g of Bacillus licheniformis.

[0072] The present application also provides a method for preparing the composite microbial additive, which specifically comprises the following steps: fully mixing the sodium butyrate, the silicon dioxide and the bran, then spray-drying the obtained mixture and the modified potato starch to form a small granular substance, and then fully mixing the small granular substance with the Clostridium butyricum, the Lactobacillus plantarum and the Bacillus licheniformis to obtain the composite microbial feed additive.

[0073] Among them, the particle size of the small granular material is 0.2-0.5mm.

[0074] In a specific embodiment, the preparation method of the above-mentioned composite microbial additive specifically comprises the following steps:

[0075] (1) Sodium butyrate, silicon dioxide and bran are fully mixed to prepare a mixture. Before mixing, the bran is crushed to meet the granulation requirements.

[0076] (2) The mixture obtained in step (1) and modified potato starch used as the wall material are spray-dried to form small granular materials of 0.2-0.5 mm.

[0077] (3) The small granular material is fully mixed with Clostridium butyricum, Lactobacillus plantarum and Bacillus licheniformis to obtain a composite microbial feed additive.

[0078] In order to make the purpose, technical scheme and advantages of the present application clearer, the technical scheme in the embodiments of the present application will be clearly and completely described below. Based on the embodiments of the present application, all other embodiments obtained by those skilled in the art without making creative work belong to the scope of protection of the present application. The embodiments described below are exemplary and are only used to explain the present application, and cannot be interpreted as limiting the present application.

[0079] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0080] In the following examples, the Clostridium butyricum, Lactobacillus plantarum and Bacillus licheniformis used are all in powder form, with a live bacteria content of 10 billion CFU / g. Meanwhile, the average particle size of the bran after crushing is less than 100 μm.

[0081] The present application is further described in detail below in conjunction with the embodiments and test results. Example Example 1

[0082] This embodiment provides a composite microbial feed additive.

[0083] The composite microbial feed additive comprises the following components in parts by weight: 20 g of sodium butyrate; 40 g of silicon dioxide; 50 g of bran; 70 g of modified potato starch; 11 g of Clostridium butyricum (with a preservation number of CGMCC NO. 27479); 6.5 g of Lactobacillus plantarum (with a preservation number of CGMCC NO. 24430); and 4 g of Bacillus licheniformis (with a preservation number of CGMCC NO. 18463).

[0084] The preparation method of the composite microbial feed additive is as follows:

[0085] (1) Sodium butyrate, silicon dioxide and bran are fully mixed to prepare a mixture; before mixing, the bran is crushed to meet granulation requirements.

[0086] (2) The mixture obtained in step (1) and modified potato starch used as the wall material are spray-dried to form small granular materials of 0.2-0.5 mm.

[0087] (3) The small granular material is fully mixed with Clostridium butyricum powder, Lactobacillus plantarum powder and Bacillus licheniformis powder to obtain a composite microbial feed additive.

[0088] Example 2-13

[0089] Examples 2-13 provide a composite microbial feed additive respectively. The difference between the above examples and Example 1 is that the addition amount of Clostridium butyricum, Lactobacillus plantarum or Bacillus licheniformis is as shown in Table 1, and the rest of the formula and preparation method are the same as those in Example 1.

[0090] The specific differences between the above embodiment and embodiment 1 are as follows:

[0091] The difference between Example 2-5 and Example 1 is: the amount of Clostridium butyricum added.

[0092] The difference between Examples 6-9 and Example 3 is the amount of Lactobacillus plantarum added.

[0093] The difference between Examples 10-13 and Example 3 is the amount of Bacillus licheniformis added. Comparative Example

[0094] Comparative Examples 1-6

[0095] Comparative Examples 1-6 provide a composite microbial feed additive respectively. The difference between the above embodiment and Example 3 is that the addition of Clostridium butyricum, Lactobacillus plantarum and Bacillus licheniformis is shown in Table 1, and the rest of the formula and preparation method are the same as those of Example 3.

[0096] Table 1 Addition of each component in the feed additive in the examples and comparative examples

[0097]

[0098] Examples 14-19

[0099] Examples 14-19 provide a composite microbial feed additive respectively. The difference between the above examples and Example 3 is that the types of Clostridium butyricum, Lactobacillus plantarum or Bacillus licheniformis are shown in Table 2, and the rest of the formula and preparation method are the same as those in Example 3.

[0100] The specific differences between the above embodiment and embodiment 3 are as follows:

[0101] The difference between Example 14-15 and Example 3 is: the type of Clostridium butyricum.

[0102] The difference between Examples 16-17 and Example 3 is the type of Lactobacillus plantarum.

[0103] The difference between Examples 18-19 and Example 3 is the type of Bacillus licheniformis.

[0104] Table 2 Types of Clostridium butyricum, Lactobacillus plantarum and Bacillus licheniformis added in the examples

[0105]

[0106] Examples 20-23

[0107] Examples 20-23 provide a composite microbial feed additive respectively. The difference between the above examples and Example 3 is that the amount of bran added is as shown in Table 3, and the rest of the formula and preparation method are the same as those in Example 3.

[0108] Comparative Examples 7-8

[0109] Comparative Examples 7-8 provide a composite microbial feed additive respectively. The difference between the above examples and Example 3 is that the amount of bran added is as shown in Table 3, and the rest of the formula and preparation method are the same as those in Example 3. Comparative Example 9

[0110] Comparative Example 9 provides a composite microbial feed additive. The difference between the above example and Example 3 is that bran is not added in this comparative example, as shown in Table 3, and the rest of the formula and preparation method are the same as those in Example 3. Comparative Example 10

[0111] Comparative Example 10 provides a feed additive. The difference between the above example and Example 3 is that Clostridium butyricum, Lactobacillus plantarum and Bacillus licheniformis are not added in this comparative example, as shown in Table 3, and the rest of the formula and preparation method are the same as those in Example 3. Comparative Example 11

[0112] Comparative Example 11 provides a composite microbial feed additive. The difference between the above example and Example 3 is that the bran is replaced by diatomaceous earth, as shown in Table 3, and the rest of the formula and preparation method are the same as those in Example 3. Comparative Example 12

[0113] Comparative Example 12 provides a composite microbial feed additive. The difference between the above embodiment and embodiment 3 is that the bran is replaced by zeolite powder, as shown in Table 3, and the rest of the formula and preparation method are the same as those of embodiment 3.

[0114] Table 3 The amount of bran added and the addition of bran and three kinds of bacteria in the examples

[0115]

[0116] Performance test results

[0117] The following tests were carried out using the composite microbial feed additives provided in the above examples and comparative examples.

[0118] 1. Broiler growth performance test

[0119] 1. Experimental Animals

[0120] 1080 one-day-old AA broiler roosters were randomly divided into 36 treatment groups, with 3 replicates in each group and 10 chickens in each replicate. Due to the limited number of broiler roosters, the experimental design for the same variable was selected to be carried out in the same batch, and the experimental design for different variables could be selected to be carried out in batches. The basic diet was: corn and soybean meal as the main raw materials, and the feed formula was designed in accordance with the Agricultural Industry Standard of the People's Republic of China for Broiler Feeding Standards (NY / T33-2004).

[0121] The diets of the 36 groups were: control group; fed with a basic diet; 23 groups were fed with a basic diet + 200 mg / kg of the composite microbial feed additive provided by Examples 1-23; and 12 groups were fed with a basic diet + 200 mg / kg of the composite microbial feed additive provided by Comparative Examples 1-12.

[0122] 2. Feeding and management

[0123] The breeding area was sanitized and disinfected 7 days before breeding, and broiler chickens were raised in conventional three-layer cages. The temperature in the chicken house was maintained at around 35°C for chicks, and the temperature in the chicken house was maintained at 22-24°C during the formal experiment. Sufficient feed and drinking water were maintained throughout the experiment, and the feed amount and feeding buckets were monitored at any time to avoid waste and inaccurate feed weighing. The chicken house was kept in good ventilation, the chickens were watched from time to time, the number of dead and sick chickens was recorded in a timely manner, those that were seriously unqualified were eliminated, and the weight of the dead chickens was recorded in a timely manner, and the feed consumption of the cage was deducted.

[0124] Immunization procedure: After one week of breeding, broiler chickens are vaccinated with Newcastle disease vaccine through drinking water method, and after two weeks, they are vaccinated with Faunal bursal disease vaccine through drinking water method, and after three weeks, they are vaccinated with Newcastle disease vaccine again through drinking water method, and at 28 days of age, Faunal bursal disease vaccine is vaccinated again through drinking water method to prevent broiler chicken Faunal bursal disease; the breeding cycle is 42 days.

[0125] 3. Index determination

[0126] (1) Production performance measurement: body weight (BWG), average daily feed intake (ADFI), average daily gain (ADG) and feed-to-gain ratio (FCR) at 42 days of age.

[0127] Average daily feed intake (ADFI) = total feed consumption / (number of experimental days × number of chickens).

[0128] Average daily gain (ADG) = (final weight / final number of chickens - initial weight / initial number of chickens) / number of experimental days.

[0129] Feed-to-gain ratio (FCR) = average daily feed intake (ADFI) / average daily gain (ADF).

[0130] Mortality rate = number of deaths / total number of animals × 100%.

[0131] (2) Data processing and analysis: All experimental data were expressed as mean ± standard deviation.

[0132] 4. Test results

[0133] The test results are shown in Table 4.

[0134] Table 4 Test results of broiler growth performance

[0135]

[0136] It can be seen from the test data in Table 4 that compared with the control group, the composite microbial feed additive provided by the present application can effectively increase the daily feed intake and daily weight gain of broilers, reduce the feed-to-weight ratio, and the weight of broilers on day 42 is significantly different from that of the control group. At the same time, compared with the control group, the composite microbial feed additive provided by the present application can also significantly reduce the mortality rate of broilers.

[0137] By comparing the test data of Example 3 and Comparative Examples 1-6, it can be seen that compared with adding only one or two of Clostridium butyricum, Lactobacillus plantarum or Bacillus licheniformis in the feed additive, adding the above three species of bacteria at the same time in the composite microbial feed additive of the present application can effectively increase the daily feed intake and daily weight gain of broilers, reduce the feed-to-weight ratio, and at the same time significantly reduce the mortality rate of broilers.

[0138] Furthermore, it can be seen from the test results of Examples 3 and 14-19 that the present application screened the strains of Clostridium butyricum, Lactobacillus plantarum or Bacillus licheniformis. From the screening results, it can be seen that when Clostridium butyricum with a preservation number of CGMCC NO. 27479, Lactobacillus plantarum with a preservation number of CGMCC NO. 24430 and Bacillus licheniformis with a preservation number of CGMCC NO. 18463 are selected, the daily feed intake and daily weight gain of broilers can be further increased, and the feed-to-weight ratio can be reduced.

[0139] In addition, by comparing the test results of Examples 3, 20-23 and Comparative Examples 7-8, it can be seen that the present application has explored the amount of bran added in the formula. When the amount of bran added is controlled at 40-60 parts, it is beneficial to increase the daily feed intake and daily weight gain of broilers and reduce the feed-to-weight ratio. When the amount of bran added is higher or lower than this range, the effects of the daily feed intake, daily weight gain and feed-to-weight ratio of broilers are far worse than the effects of the daily feed intake, daily weight gain and feed-to-weight ratio of broilers when the amount of bran added is controlled at 40-60 parts.

[0140] At the same time, through the comparison between Example 3 and Comparative Examples 9-10, it can be seen that the daily feed intake, daily weight gain and feed-to-weight ratio of broilers when only three strains or bran are added are far worse than the daily feed intake, daily weight gain and feed-to-weight ratio of broilers when three strains and bran are added at the same time. And when the bran is replaced with diatomaceous earth or zeolite powder, the daily feed intake, daily weight gain and feed-to-weight ratio of broilers are also not as good as the daily feed intake, daily weight gain and feed-to-weight ratio of broilers when three strains and bran are added at the same time. It can be seen that in the scheme of the present application, the simultaneous addition of bran and three strains can further effectively increase the daily feed intake and daily weight gain of broilers and reduce the feed-to-weight ratio.

[0141] (II) Broiler intestinal repair test

[0142] The normal structure and function of the intestine are the basic guarantee for the adequate digestion and absorption of nutrients. In particular, the height of the intestinal villi, the depth of the crypts, and V / C are important indicators for measuring the digestion and absorption function of the intestine.

[0143] The height of intestinal villi is significantly correlated with the number of cells. In the finger-shaped villi, the height of the villi is related to the number of intestinal epithelial cells. When the villi become longer, the number of intestinal epithelial cells increases; when the villi become shorter, the number of mature intestinal epithelial cells decreases, and the ability to absorb nutrients is low.

[0144] Crypt depth reflects the rate of cell generation, as cells constantly migrate and differentiate from the base of the crypt to the end of the villus to replenish the normal shedding of the villus epithelium. Therefore, the crypt can be considered a villus processing factory. If this process slows down, the rate of cell generation at the base decreases, making the crypt shallower. Shallowing of the crypt indicates an increase in the maturation rate of intestinal epithelial cells and enhanced absorptive function. Deepening of the crypt indicates accelerated tissue metabolism and an increased need for new tissue, which means an increase in the nutrients required for maintenance and a decrease in animal production efficiency.

[0145] V / C reflects the functional state of the intestine. When the ratio increases, the mucosa improves, the digestive and absorption functions are enhanced, and the growth and development are accelerated; when the ratio decreases, the mucosa is damaged and the digestive and absorption functions are reduced.

[0146] The intestinal tract (duodenum) of broilers fed with the above-mentioned Example 3, Comparative Examples 4-6 and the control group was tested respectively. The test indicators included intestinal villus height (V), crypt depth (C) and V / C. The number of broilers tested in each group was 20.

[0147] The test results are shown in Table 5.

[0148] Table 5 Test results of broiler growth performance

[0149]

[0150] As shown in Table 5, compared with the test results of the control group and comparative examples 4-6, the composite microbial feed additive of Example 3 can increase the height of the intestinal villi of broilers by 19.5% and reduce the depth of the intestinal crypts of broilers to 58.5% when the broilers are marketed for 42 days, significantly improving V / C. Therefore, the feed additive provided by the present application can effectively promote the development of the broiler intestine and improve the absorption of nutrients.

[0151] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example" or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0152] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present application, rather than to limit it. Although the present application has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A composite microbial feed additive, characterized in that: The composite microbial feed additive comprises the following components in parts by weight: 10-30 parts of sodium butyrate; 30-50 parts of silicon dioxide; 40-60 parts of bran; 60-80 parts of modified potato starch; 10-12 parts of Clostridium butyricum; 5.5-7.5 parts of Lactobacillus plantarum; and 2.5-5.5 parts of Bacillus licheniformis. The deposit number of the Clostridium butyricum is CGMCC NO.27479; The deposit number of the Lactobacillus plantarum is CGMCC NO.24430; The deposit number of the Bacillus licheniformis is CGMCC NO.18463.

2. The composite microbial feed additive according to claim 1, characterized in that: The weight portion of the bran is 45-55 parts.

3. A method for preparing the composite microbial feed additive according to any one of claims 1 to 2, characterized in that: The preparation method specifically comprises the following steps: fully mixing the sodium butyrate, the silicon dioxide and the bran, then spray-drying the obtained mixture and the modified potato starch to form small granular substances, and then fully mixing the small granular substances with the Clostridium butyricum, the Lactobacillus plantarum and the Bacillus licheniformis to obtain the composite microbial feed additive.

4. The method for preparing the composite microbial feed additive according to claim 3, characterized in that: The particle size of the small granular material is 0.2-0.5 mm.

5. The method for preparing the composite microbial feed additive according to claim 3, characterized in that: The Clostridium butyricum, the Lactobacillus plantarum and the Bacillus licheniformis are all in powder form.

6. A feed, characterized in that: The feed comprises the composite microbial feed additive according to any one of claims 1-2.

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