Livestock composite biological feed prepared from probiotics and method thereof

Abstract

The invention discloses a livestock composite biological feed prepared from probiotics and a method thereof, and relates to the technical field of livestock feeds. The compound biological feed for the livestock and poultry is prepared from the following raw materials: lactobacillus plantarum BNCC138404, bacillus coagulans BNCC132591, bacillus velezensis BNCC352076, bifidobacterium bifidum BNCC186493, aspergillus niger BNCC147472, enterococcus faecalis BNCC186075, yarrowia lipolytica BNCC193899, nutrient substances, mineral trace elements, vitamins, amino acids and prebiotics. The compound biological feed for the livestock and poultry is prepared from the following raw materials: lactobacillus plantarum BNCC138404, bacillus coagulans BNCC132591, bacillus velezensis BNCC352076, bifidobacterium bifidum BNCC186493, aspergillus The compound biological feed fermented by adopting various probiotics can improve the antioxidant level of the broiler chicken, improve the health condition of intestinal tracts, increase the villus height and the villus-implicit ratio of the jejunum of the broiler chicken and reduce the occurrence of intestinal inflammation, so that the growth performance of the broiler chicken is promoted, and the immune performance of the body is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of livestock and poultry feed, and more specifically to a composite biological feed for livestock and poultry prepared by using probiotics and a method thereof. Background Art

[0002] With the increase in the scale and intensiveness of the breeding industry, the production performance of animals and the output of livestock products have been significantly improved, but many problems have also come with it. With the increase in breeding density, the stress level of animals also increases, the immune function of animals decreases, and the resistance to various harmful microorganisms becomes lower and lower, making animals in a sub-healthy state for a long time and easily infected with various diseases. High-density breeding also seriously deteriorates the breeding environment. In actual production, antibiotics are added to feed to prevent diseases and improve economic benefits. However, long-term consumption of feed with antibiotics by animals will bring many adverse consequences, such as bacterial resistance, affecting the microecological balance and body immunity of animals, and the antibiotics accumulated in large quantities in animals are enriched in the human body through the dining table, affecting human health.

[0003] Compound biological feed is a new type of fermented biological feed that uses beneficial microorganisms to ferment one or more feed raw materials anaerobic or aerobic, which can improve the utilization rate of nutrients in feed raw materials, eliminate anti-nutritional factors, and at the same time, the beneficial microorganisms multiply and enrich, and functional primary and secondary metabolites are generated in large quantities. It contains rich antimicrobial active substances, probiotics, and nutritional factors, which can promote digestion and absorption of farmed animals, reduce feed-to-meat ratio, enhance body resistance, and inhibit the growth of pathogens. In recent years, it has become the preferred option to replace antibiotics.

[0004] Since the quality of compound biological feed is directly related to the quality of biological feed, the quality of biological feed is uneven at present, the combination of probiotic species is unreasonable, the function of bacteria is single, and the microbial fermentation and decomposition ability is poor, which seriously affects the quality of biological feed. Therefore, there is still a need to develop compound biological feed with comprehensive functions and good growth-promoting effects. Summary of the invention

[0005] In view of this, the present invention provides a composite biological feed for poultry and livestock prepared by using probiotics and a method thereof.

[0006] In order to achieve the above object, the present invention adopts the following technical solution:

[0007] A compound biological feed for poultry and livestock prepared using probiotics, characterized in that the raw materials of the compound biological feed for poultry and livestock include: Lactobacillus plantarum BNCC138404, Bacillus coagulans BNCC132591, Bacillus velez BNCC352076, Bifidobacterium bifidum BNCC186493, Aspergillus niger BNCC147472, Enterococcus faecalis BNCC186075 and Yarrowia lipolytica BNCC193899.

[0008] Preferably, the bacterial contents of Lactobacillus plantarum, Bacillus coagulans, Bacillus velez, Bifidobacterium bifidum, Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica are 1-2×10 8 cfu / mL, 3-4×10 8 cfu / mL, 2-3×10 8 cfu / mL, 5-8×10 8 cfu / mL, 1-3×10 8 cfu / mL, 2-3×10 8 cfu / mL and 2-3×10 8 cfu / mL.

[0009] Furthermore, the raw materials of the compound biological feed for poultry and livestock also include nutrients, mineral trace elements, vitamins, amino acids and prebiotics.

[0010] Preferably, the nutrients include soybean meal, corn, wheat bran and grass meal.

[0011] Preferably, the mineral trace elements include copper sulfate, manganese sulfate, zinc oxide, ferrous sulfate, sodium selenite, calcium iodate and cobalt chloride.

[0012] Preferably, the vitamins include vitamin A, vitamin D, vitamin E, vitamin B1, vitamin B3, vitamin B6, vitamin B 12 , folic acid and niacin.

[0013] Preferably, the amino acids include methionine, threonine and lysine.

[0014] Preferably, the prebiotics include soybean oligosaccharides, mannooligosaccharides and galactooligosaccharides.

[0015] Furthermore, the weight-to-volume ratio of the Lactobacillus plantarum, Bacillus coagulans, Bacillus velezensis, Bifidobacterium bifidum, Aspergillus niger, Enterococcus faecalis, Yarrowia lipolytica, nutrients, mineral trace elements, vitamins, amino acids and prebiotics is (3-7)L:(3-7)L:(2-6)L:(1-5)L:(1-3)L:(0.5-3)L:(0.5-3.5)L:(15-25)kg:(1-5)kg:(1-3)kg:(1-3)kg:(0.2-2)kg.

[0016] As the same inventive concept as the above technical solution, the present invention also claims protection for the above method for preparing composite biological feed for poultry and livestock using probiotics, which is characterized by comprising the following steps:

[0017] (1) removing impurities from the nutrients, crushing them, passing them through a 40-60 mesh sieve, and adding sterile water to obtain a liquid raw material culture medium with a water content of 50%-60%;

[0018] (2) activating Lactobacillus plantarum, Bacillus coagulans, Bacillus velezensis, Bifidobacterium bifidum, Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica respectively, and then inoculating them into seed culture medium for cultivation to obtain seed solutions of different strains;

[0019] (3) mixing the seed liquids of different strains to obtain a composite strain seed liquid, then transferring the mixture to a liquid raw material culture medium, stirring and mixing the mixture, adjusting the water content, and then performing a first fermentation to obtain a first fermentation product;

[0020] (4) adding mineral trace elements, vitamins, amino acids and prebiotics to the first fermentation product and stirring evenly, and then performing a second fermentation. After the fermentation is completed, a composite biological feed for livestock and poultry can be obtained.

[0021] Furthermore, in step (2), the preparation method of the Lactobacillus plantarum, Bacillus coagulans, Bacillus velez and Bifidobacterium bifidum seed liquid is: Lactobacillus plantarum, Bacillus coagulans, Bacillus velez and Bifidobacterium bifidum strains are respectively inoculated on a slant culture medium for activation, and then the activated strains are inoculated into a liquid culture medium with an inoculation loop for cultivation, to obtain the seed liquid of Lactobacillus plantarum, Bacillus coagulans, Bacillus velez and Bifidobacterium bifidum, respectively.

[0022] Preferably, the conditions for the seed activation culture are: culture temperature range 28-32° C., culture 36-48 h.

[0023] Preferably, the culture conditions of the seed solution are: culture temperature range 28-32°C, stirring speed 150-200rpm / min, and culture 12-24h.

[0024] Furthermore, in step (2), the preparation method of the Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica seed liquid is: inoculate the Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica strains onto a slant culture medium for activation, and then inoculate the activated strains into a liquid culture medium with an inoculation loop for cultivation, to obtain the Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica seed liquids, respectively.

[0025] Preferably, the conditions for the seed activation culture are: culture temperature range 24-26°C, culture 24-36h.

[0026] Preferably, the culture conditions of the seed solution are: culture temperature range 24-26°C, stirring speed 220-270rpm / min, and culture 24-36h.

[0027] Furthermore, in step (1)-step (2), the slant culture medium is prepared as follows: 10 g of peptone, 10 g of yeast powder, 20 g of glucose, 20 g of agar, add water to make up to 1000 mL, pH value 7.0, and sterilize at 121°C for 20 min.

[0028] Furthermore, the preparation method of the liquid culture medium is: 10g of peptone, 10g of yeast powder, 20g of glucose, 5g of sodium chloride, adding water to make up to 1000mL, pH value 7.0, and sterilizing at 121°C for 20min.

[0029] Furthermore, in step (3), the water content is 40-50%; the inoculation amount of the composite bacterial seed liquid is 3-5%; the first fermentation is aerobic fermentation, the fermentation temperature is 20-30°C, and the fermentation time is 4-8d.

[0030] Furthermore, in step (4), the second fermentation is anaerobic fermentation, the fermentation temperature is 35-45° C., and the fermentation time is 10-18 days.

[0031] As an inventive concept identical to the above technical solution, the present invention also requests protection for the application of the above compound biological feed for livestock and poultry in breeding livestock and poultry.

[0032] Preferably, the compound biological feed for poultry and livestock improves growth performance.

[0033] Preferably, the compound biological feed for poultry and livestock improves the immune performance of the body.

[0034] Preferably, the compound biological feed for poultry and livestock improves intestinal health.

[0035] It can be seen from the above technical solutions that, compared with the prior art, the present invention discloses a composite biological feed for livestock and poultry prepared by using probiotics and a method thereof, which has the following beneficial effects:

[0036] The present invention adopts a composite bacterial liquid fermentation feed prepared by multiple probiotics (Lactobacillus plantarum, Bacillus coagulans, Bacillus velez, Bifidobacterium bifidum, Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica), wherein Lactobacillus plantarum is a kind of lactic acid bacteria, which can produce lactobacillus or ecological niche competition to inhibit intestinal pathogens, and can prevent cell apoptosis, enhance intestinal function and form a biological protection barrier; Bacillus can promote the proliferation of beneficial bacteria, inhibit the colonization of pathogenic bacteria, thereby maintaining the balance of intestinal flora, and at the same time can improve feed utilization and improve Intestinal mucosal structure, thereby improving intestinal digestibility and promoting animal growth; Aspergillus niger can secrete more cellulase, hemicellulase and protease to improve feed utilization; Enterococcus faecalis can produce natural antibiotics, which is beneficial to the health of the body. At the same time, it can also produce antibacterial substances such as bacteriocins to inhibit the growth of pathogens such as Escherichia coli and Salmonella, and improve the intestinal microenvironment; in addition to degrading macromolecular substances in feed raw materials, yeast can also provide growth factors for other compatible probiotics, thereby improving the decomposition efficiency and conversion efficiency of feed.

[0037] The composite biological feed fermented by various probiotic strains can improve the antioxidant level of broilers, improve intestinal health, increase the villus height and villus-to-crypt ratio of the jejunum of broilers, and reduce the occurrence of intestinal inflammation, thereby promoting the growth performance of broilers and improving the immune performance of the body. DETAILED DESCRIPTION

[0038] The following will be described clearly and completely in conjunction with the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0039] Lactobacillus plantarum, Bacillus coagulans, Bacillus velez, Bifidobacterium bifidum, Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica in the following examples were purchased from Beijing Kezhan Biotechnology Co., Ltd. with product numbers BNCC138404, BNCC132591, BNCC352076, BNCC186493, B NCC147472, BNCC186075 and BNCC193899, respectively.

[0040] The culture medium formula used in the following examples is as follows:

[0041] Slant culture medium: peptone 10g, yeast powder 10g, glucose 20g, agar 20g, add water to 1000mL, pH 7.0, sterilize at 121℃ for 20min.

[0042] Liquid culture medium: peptone 10g, yeast powder 10g, glucose 20g, sodium chloride 5g, add water to 1000mL, pH 7.0, sterilize at 121℃ for 20min.

[0043] Example 1

[0044] A method for preparing composite biological feed for poultry and livestock using probiotics, characterized by comprising the following steps:

[0045] (1) 5 kg of soybean meal, 5 kg of corn, 3 kg of wheat bran, and 2 kg of grass powder were crushed after removing impurities, passed through a 40-mesh sieve, mixed evenly, and sterile water was added to obtain a liquid raw material culture medium with a water content of 50%;

[0046] (2) preparing Lactobacillus plantarum, Bacillus coagulans, Bacillus velezii, and Bifidobacterium bifidum seed solutions respectively: inoculating Lactobacillus plantarum, Bacillus coagulans, Bacillus velezii, and Bifidobacterium bifidum strains onto slant culture medium respectively, activating and culturing at 28° C. for 48 h, then inoculating the activated strains into liquid culture medium using an inoculation loop, and culturing at 28° C. and 150 rpm for 24 h, thereby obtaining Lactobacillus plantarum seed solution, Bacillus coagulans seed solution, Bacillus velezii seed solution, and Bifidobacterium bifidum seed solution respectively;

[0047] (3) preparing seed solutions of Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica respectively: inoculating Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica strains on slant culture medium respectively, activating and culturing at 24° C. for 36 h, and then inoculating the activated strains into liquid culture medium with an inoculation loop respectively, culturing at 24° C. and 270 rpm for 24 h, thereby obtaining Aspergillus niger seed solution, Enterococcus faecalis seed solution and Yarrowia lipolytica seed solution respectively;

[0048] (4) 3 L of Lactobacillus plantarum seed liquid, 5 L of Bacillus coagulans seed liquid, 4 L of Bacillus velez seed liquid, 1 L of Bifidobacterium bifidum seed liquid, 1 L of Aspergillus niger seed liquid, 1.5 L of Enterococcus faecalis seed liquid and 2 L of Yarrowia lipolytica seed liquid were mixed to obtain a composite strain seed liquid, and then the composite strain seed liquid was transferred to a liquid raw material culture medium at an inoculation amount of 3%, stirred and mixed, and the water content was adjusted to 40%, and then aerobic fermentation was carried out at 25° C. for 6 days to obtain a first fermentation product;

[0049] (4) Add 3 kg of mineral trace elements, 2 kg of vitamins, 2 kg of amino acids and 1.3 kg of prebiotics to the first fermentation product and mix well, then perform anaerobic fermentation at 35°C for 18 days. After the fermentation is completed, the livestock and poultry composite biological feed can be obtained.

[0050] Example 2

[0051] A method for preparing composite biological feed for poultry and livestock using probiotics, characterized by comprising the following steps:

[0052] (1) 7 kg of soybean meal, 6 kg of corn, 4 kg of wheat bran, and 3 kg of grass powder were crushed after removing impurities, passed through a 50-mesh sieve, mixed evenly, and sterile water was added to obtain a liquid raw material culture medium with a water content of 55%;

[0053] (2) preparing Lactobacillus plantarum, Bacillus coagulans, Bacillus velezii, and Bifidobacterium bifidum seed solutions respectively: inoculating Lactobacillus plantarum, Bacillus coagulans, Bacillus velezii, and Bifidobacterium bifidum strains onto slant culture medium respectively, culturing at 30° C. for 42 h, then inoculating the activated strains into liquid culture medium using an inoculation loop, culturing at 30° C. and 175 rpm for 30 h, thereby obtaining Lactobacillus plantarum seed solution, Bacillus coagulans seed solution, Bacillus velezii seed solution, and Bifidobacterium bifidum seed solution respectively;

[0054] (3) preparing seed solutions of Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica respectively: inoculating Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica strains on slant culture medium respectively, activating and culturing at 25° C. for 30 h, and then inoculating the activated strains into liquid culture medium with an inoculation loop respectively, and culturing at 25° C. and 220 rpm for 36 h, thereby obtaining Aspergillus niger seed solution, Enterococcus faecalis seed solution and Yarrowia lipolytica seed solution respectively;

[0055] (4) 5 L of Lactobacillus plantarum seed liquid, 3 L of Bacillus coagulans seed liquid, 6 L of Bacillus velez seed liquid, 3 L of Bifidobacterium bifidum seed liquid, 2 L of Aspergillus niger seed liquid, 0.5 L of Enterococcus faecalis seed liquid and 3.5 L of Yarrowia lipolytica seed liquid were mixed to obtain a composite strain seed liquid, and then the composite strain seed liquid was transferred to a liquid raw material culture medium at an inoculation amount of 3%, stirred and mixed, and the water content was adjusted to 45%, and then aerobic fermentation was carried out at 30° C. for 4 days to obtain a first fermentation product;

[0056] (4) Add 5 kg of mineral trace elements, 3 kg of vitamins, 1 kg of amino acids and 2 kg of prebiotics to the first fermentation product and mix well, then perform anaerobic fermentation at 45°C for 10 days. After the fermentation is completed, the livestock and poultry composite biological feed can be obtained.

[0057] Example 3

[0058] A method for preparing composite biological feed for poultry and livestock using probiotics, characterized by comprising the following steps:

[0059] (1) 7 kg of soybean meal, 6 kg of corn, 7 kg of wheat bran, and 5 kg of grass powder were crushed after removing impurities, passed through a 60-mesh sieve, mixed evenly, and sterile water was added to obtain a liquid raw material culture medium with a water content of 60%;

[0060] (2) preparing Lactobacillus plantarum, Bacillus coagulans, Bacillus velezii, and Bifidobacterium bifidum seed solutions respectively: inoculating Lactobacillus plantarum, Bacillus coagulans, Bacillus velezii, and Bifidobacterium bifidum strains onto slant culture medium respectively, activating and culturing at 32° C. for 36 h, then inoculating the activated strains into liquid culture medium using an inoculation loop, culturing at 32° C. and 200 rpm for 12 h, thereby obtaining Lactobacillus plantarum seed solution, Bacillus coagulans seed solution, Bacillus velezii seed solution, and Bifidobacterium bifidum seed solution respectively;

[0061] (3) preparing seed solutions of Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica respectively: inoculating Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica strains on slant culture medium respectively, activating and culturing at 26° C. for 24 h, and then inoculating the activated strains into liquid culture medium using an inoculation loop respectively, and culturing at 26° C. and 245 rpm for 30 h, thereby obtaining Aspergillus niger seed solution, Enterococcus faecalis seed solution and Yarrowia lipolytica seed solution respectively;

[0062] (4) 7 L of Lactobacillus plantarum seed liquid, 7 L of Bacillus coagulans seed liquid, 2 L of Bacillus velez seed liquid, 5 L of Bifidobacterium bifidum seed liquid, 1 L of Aspergillus niger seed liquid, 3 L of Enterococcus faecalis seed liquid and 0.5 L of Yarrowia lipolytica seed liquid were mixed to obtain a composite strain seed liquid, and then the composite strain seed liquid was transferred to a liquid raw material culture medium at an inoculation amount of 3%, stirred and mixed, and the water content was adjusted to 40%, and then aerobic fermentation was carried out at 25° C. for 6 days to obtain a first fermentation product;

[0063] (4) Add 1 kg of mineral trace elements, 1 kg of vitamins, 3 kg of amino acids and 0.2 kg of prebiotics to the first fermentation product and mix well, then perform anaerobic fermentation at 35°C for 18 days. After the fermentation is completed, the livestock and poultry composite biological feed can be obtained.

[0064] Comparative Example 1

[0065] The difference from Example 2 is that probiotics such as Lactobacillus plantarum, Bacillus coagulans, Bacillus velez, Bifidobacterium bifidum, Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica are removed.

[0066] Comparative Example 2

[0067] The difference from Example 2 is that the prebiotics are removed.

[0068] Comparative Example 3

[0069] The difference from Example 2 is that in step (4)-step (5), the composite bacterial seed liquid is transferred to the liquid raw material culture medium and stirred and mixed, and then mineral trace elements, vitamins, amino acids and prebiotics are added, and fermented at 30°C for 30 days to obtain the livestock and poultry composite biological feed.

[0070] Experimental Example 1 Effects of different compound biological feeds on the growth performance of broiler chickens

[0071] The experiment selected 210 1-day-old broilers with similar weight and health, and randomly divided them into 7 groups, 30 in each group, one of which was fed with a basic diet as a control (the specific composition is shown in Table 1), and the remaining 6 groups were fed with the composite biological feed prepared in Examples 1-3 and Comparative Examples 1-3, respectively. The breeding method was cage breeding. Before the experiment, the chicken house, chicken cages, chicken troughs, etc. were cleaned and disinfected, and free access to food, drinking water and ventilation were allowed. During the breeding process, the chickens were weighed at 14, 28, and 42 days of age, and the average weight of the broilers at each stage was recorded. The feed-to-meat ratio of the broilers was calculated on the day of market release, and the measurement results are shown in Table 2.

[0072] Table 1 Basic feed composition and nutritional level

[0073]

[0074]

[0075] Table 2 Effects of different compound biological feeds on broiler growth performance

[0076]

[0077] As can be seen from Table 2, compared with the control group, the composite biological feed provided by Examples 1-3 of the present invention has a good growth-promoting effect on broilers and can reduce the feed-to-meat ratio. As can be seen from Comparative Example 1, although the composite biological feed after removing the probiotics can also promote the growth of broilers, its effect is significantly lower than that of the control group and Examples 1-3, and the feed-to-meat ratio is also higher than that of the present invention. It can be seen that the use of probiotics to ferment feed can increase the nutrients in the biological feed, enhance the growth performance of broilers, and also reduce the feed-to-meat ratio; as can be seen from Comparative Example 2, although its effect after removing the prebiotics is lower than that of the present invention, it is higher than Comparative Example 1. It can be seen that when probiotics and prebiotics are used together, the growth performance of broilers can be enhanced. Compared with Comparative Example 3, the method of alternating aerobic and anaerobic fermentation in Examples 1-3 can be beneficial to the survival of microorganisms, the growth of metabolites and nutrients, and thus can further promote the growth performance of broilers.

[0078] Experimental Example 2 Effects of different compound biological feeds on broiler immune organ indexes

[0079] Ten broilers were randomly selected from each group at 14 and 28 days of age, weighed, bled by carotid artery, and then dissected under sterile conditions to separate the spleen, thymus, and bursa of Fabricius. The fat was removed and weighed, and the immune organ index was calculated. The test results are shown in Table 3.

[0080] Table 3 Effects of different compound biological feeds on broiler immune organ indexes

[0081]

[0082] The bursa of Fabricius is a humoral immune organ unique to poultry; the thymus is an important lymphatic organ of the body, and its function is closely related to immunity. It is the place where T cells differentiate, develop, and mature; and the spleen is the largest peripheral immune organ of poultry; in the absence of infection, the development state of the immune organ determines the immune level. As shown in Table 3, compared with the control group, the composite biological feed prepared by Examples 1-3 of the present invention can significantly improve the spleen, thymus and bursa of Fabricius index of broilers, among which Example 2 shows better immune performance; it shows that the composite biological feed of the present invention has a good promoting effect on the development of broiler immune organs; and after the probiotics are removed from Comparative Example 1, it reduces the spleen, thymus and bursa of broilers compared with the control group, and cannot promote the development of broiler immune organs; and although Comparative Example 2-3 groups can promote the development of immune organs, the difference with the control group is not obvious.

[0083] Experimental Example 3 Effects of different compound biological feeds on T-AOC, MDA and globulin content in broiler serum

[0084] Blood was collected from the carotid artery of broilers at 14 and 28 days of age, respectively, and the serum was separated by centrifugation at 3000 r / min for 10 min. The content was detected using T-AOC, MDA, and globulin kits. The measurement results are shown in Table 4.

[0085] Table 4 Effects of different compound biological feeds on T-AOC, MDA and globulin content in broiler serum

[0086]

[0087] As shown in Table 4, compared with the control group, the composite biological feed prepared by Examples 1-3 of the present invention can significantly increase the T-AOC level and globulin content, and reduce the MDA level, among which Example 2 shows better immune performance; since T-AOC is an indicator for measuring the body's antioxidant capacity, its level represents the body's total antioxidant capacity, MDA is one of the most important products of peroxidation, and its production can aggravate the damage of cell membranes, and the level of globulin is an important indicator of the body's immune performance. It can be shown that the composite biological feed of the present invention can effectively improve the antioxidant capacity and immune performance of broilers; while the comparative examples 1-3 groups increased T-AOC and globulin and reduced MDA compared with the control group, but the effect was significantly lower than that of the present invention.

[0088] Experimental Example 4 Effects of different compound biological feeds on intestinal health performance of broiler chickens

[0089] After the experiment, 10 broilers were randomly selected from each treatment for slaughter (fasting in advance), and about 2 cm of jejunal tissue was collected. After being rinsed with saline, it was fixed with 10% formalin for 24 hours and then stained with hematoxylin-eosin to make paraffin sections. The villus height and crypt depth were measured under a microscope, and the villus crypt ratio (villus height / crypt depth) was calculated. A 2 cm sample from the middle jejunum was collected and rinsed with saline. Intestinal inflammation was scored, which was divided into 2 levels, with level 0 indicating no damage visible to the naked eye; level 1 indicating congestion but no necrosis; level 2 indicating necrosis or ulceration; the measurement results are shown in Table 5.

[0090] Table 5 Determination of the effects of different compound biological feeds on the intestinal health of broiler chickens

[0091] Group Villus height / μm Crypt depth / μm Velvet ratio Intestinal inflammation Control group 988.26 156.07 6.33 1.61 Example 1 1041.83 154.29 6.75 1.14 Example 2 1048.27 153.17 6.84 1.02 Example 3 1037.91 153.53 6.76 1.16 Comparative Example 1 1001.23 154.58 6.48 1.52 Comparative Example 2 1016.28 153.88 6.60 1.43 Comparative Example 3 1024.04 153.72 6.66 1.35

[0092] As shown in Table 5, compared with the control group, the composite biological feed prepared by Examples 1-3 of the present invention can improve the villus height and villus-crypt ratio of broilers after feeding them, which are increased by 5.15% and 6.22%, 5.72% and 7.46%, 4.78% and 6.36%, respectively. There is no significant difference in the crypt depth compared with the control group, and the intestinal inflammation of the jejunum of broilers is significantly reduced, which is reduced by 29.19%, 35.40% and 27.95%, respectively; while the comparative examples 1-3 groups have improved the villus height and villus-crypt ratio compared with the control group, but the improvement effect is significantly lower than that of the present invention. It can be shown that the composite biological feed fermented by probiotics in the present invention can significantly promote the intestinal health of broilers, improve the villus height and villus-crypt ratio of the jejunum of broilers, and reduce the occurrence of intestinal inflammation; the higher the villus height, the larger the intestinal absorption area, and the higher the villus-crypt ratio, the stronger the digestion and absorption capacity.

[0093] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0094] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A compound biological feed for poultry and livestock prepared using probiotics, characterized in that: The raw materials of the poultry and livestock compound biological feed include: Lactobacillus plantarum BNCC138404, Bacillus coagulans BNCC132591, Bacillus velez BNCC352076, Bifidobacterium bifidum BNCC186493, Aspergillus niger BNCC147472, Enterococcus faecalis BNCC186075 and Yarrowia lipolytica BNCC193899; Preferably, the bacterial contents of Lactobacillus plantarum, Bacillus coagulans, Bacillus velez, Bifidobacterium bifidum, Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica are 1-2×10 8 cfu / mL, 3-4×10 8 cfu / mL, 2-3×10 8 cfu / mL, 5-8×10 8 cfu / mL, 1-3×10 8 cfu / mL, 2-3×10 8 cfu / mL and 2-3×10 8 cfu / mL.

2. The compound biological feed for poultry and livestock according to claim 1, characterized in that: The raw materials of the compound biological feed for poultry and livestock also include nutrients, mineral trace elements, vitamins, amino acids and prebiotics; Preferably, the nutrients include soybean meal, corn, wheat bran and grass meal; Preferably, the mineral trace elements include copper sulfate, manganese sulfate, zinc oxide, ferrous sulfate, sodium selenite, calcium iodate and cobalt chloride; Preferably, the vitamins include vitamin A, vitamin D, vitamin E, vitamin K, vitamin B1, vitamin B3, vitamin B6, vitamin B 12 , folic acid, and niacin; Preferably, the amino acids include methionine, threonine and lysine; Preferably, the prebiotics include soybean oligosaccharides, mannooligosaccharides and galactooligosaccharides.

3. The compound biological feed for poultry and livestock according to claim 1, characterized in that: The weight-to-volume ratio of the Lactobacillus plantarum, Bacillus coagulans, Bacillus velezensis, Bifidobacterium bifidum, Aspergillus niger, Enterococcus faecalis, Yarrowia lipolytica, nutrients, mineral trace elements, vitamins, amino acids and prebiotics is (3-7) L: (3-7) L: (2-6) L: (1-5) L: (1-3) L: (0.5-3) L: (0.5-3.5) L: (15-25) kg: (1-5) kg: (1-3) kg: (1-3) kg: (0.2-2) kg.

4. A method for preparing composite biological feed for poultry and livestock using probiotics according to any one of claims 1 to 3, characterized in that: The following steps are involved: (1) removing impurities from the nutrients, crushing them, passing them through a 40-60 mesh sieve, and adding sterile water to obtain a liquid raw material culture medium with a water content of 50%-60%; (2) activating Lactobacillus plantarum, Bacillus coagulans, Bacillus velezensis, Bifidobacterium bifidum, Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica respectively, and then inoculating them into seed culture medium for cultivation to obtain seed solutions of different strains; (3) mixing the seed liquids of different strains to obtain a composite strain seed liquid, then transferring the mixture to a liquid raw material culture medium, stirring and mixing the mixture, adjusting the water content, and then performing a first fermentation to obtain a first fermentation product; (4) adding mineral trace elements, vitamins, amino acids and prebiotics to the first fermentation product and stirring evenly, and then performing a second fermentation. After the fermentation is completed, a composite biological feed for livestock and poultry can be obtained.

5. The preparation method according to claim 4, characterized in that: In step (2), the preparation method of the Lactobacillus plantarum, Bacillus coagulans, Bacillus velez and Bifidobacterium bifidum seed liquid is as follows: Lactobacillus plantarum, Bacillus coagulans, Bacillus velez and Bifidobacterium bifidum strains are inoculated on a slant culture medium for activation, and then the activated strains are inoculated into a liquid culture medium with an inoculation loop for cultivation, respectively, to obtain Lactobacillus plantarum, Bacillus coagulans, Bacillus velez and Bifidobacterium bifidum seed liquids; Preferably, the conditions for the seed activation culture are: culture temperature range 28-32°C, culture 36-48h; Preferably, the culture conditions of the seed solution are: culture temperature range 28-32°C, stirring speed 150-200rpm / min, and culture 12-24h.

6. The preparation method according to claim 4, characterized in that: In step (2), the preparation method of the Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica seed liquid is: inoculate Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica strains on a slant culture medium for activation, and then inoculate the activated strains into a liquid culture medium with an inoculation loop for cultivation, respectively, to obtain the Aspergillus niger, Enterococcus faecalis and Yarrowia lipolytica seed liquid; Preferably, the conditions for the seed activation culture are: a culture temperature range of 24-26° C., and a culture time of 24-36 h; Preferably, the culture conditions of the seed solution are: culture temperature range 24-26°C, stirring speed 220-270rpm / min, and culture 24-36h.

7. The preparation method according to claim 4, characterized in that: In step (1)-step (2), the slant culture medium is prepared as follows: 10 g of peptone, 10 g of yeast powder, 20 g of glucose, 20 g of agar, add water to make up to 1000 mL, pH value 7.0, and sterilize at 121°C for 20 min; The liquid culture medium is prepared as follows: 10 g of peptone, 10 g of yeast powder, 20 g of glucose, 5 g of sodium chloride, water is added to make the volume to 1000 mL, the pH value is 7.0, and sterilized at 121° C. for 20 min.

8. The preparation method according to claim 4, characterized in that: In step (3), the water content is 40-50%; the inoculation amount of the composite bacterial seed liquid is 3-5%; the first fermentation is aerobic fermentation, the fermentation temperature is 20-30°C, and the fermentation time is 4-8d.

9. The preparation method according to claim 4, characterized in that: In step (4), the second fermentation is anaerobic fermentation, the fermentation temperature is 35-45° C., and the fermentation time is 10-18 days.

10. Use of the compound biological feed for livestock and poultry according to any one of claims 1 to 3 in breeding livestock and poultry; Preferably, the compound biological feed for poultry and livestock improves growth performance; Preferably, the poultry and livestock compound biological feed improves the body's immune performance; Preferably, the compound biological feed for poultry and livestock improves intestinal health.

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