Processing technology method of fermented bran with high lactic acid bacteria number

Abstract

The invention discloses a processing technology method of fermented bran with a high lactic acid bacteria number, and belongs to the technical field of biology. Comprising the following steps: taking sterilized bran as a substrate, taking amino acid, yeast extract powder, a compound enzyme preparation, mineral ions and inorganic salt as auxiliary materials, inoculating strains, then carrying out solid anaerobic fermentation, and then carrying out low-temperature drying to generate a product. According to the method, processing and value increasing of the wheat bran can be achieved at low cost, the nutritional value, digestibility and oxidation resistance of the wheat bran are improved, antinutritional factors are reduced, and the food calling performance and the probiotic effect are improved.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a processing method of high lactic acid bacteria count fermented bran. Background Art

[0002] Lactic acid bacteria are common intestinal bacteria that can improve gastrointestinal function and enhance the body's immune capacity. After livestock and poultry take lactic acid bacteria, lactic acid bacteria tightly bind to intestinal mucosal cells through adhesins, colonize on the surface of the intestinal mucosa, and become the main component of the physiological barrier, thereby restoring host resistance, repairing the intestinal flora barrier, and curing intestinal diseases. Lactic acid bacteria can colonize in the intestine, which is equivalent to natural automatic immunity. Lactic acid bacteria can significantly activate the phagocytic function of macrophages, stimulate peritoneal macrophages, induce the production of interferon, promote cell division, produce antibodies, and promote cellular immunity, so lactic acid bacteria have the ability to enhance the body's nonspecific and specific immune responses and improve the body's disease resistance.

[0003] In addition, lactic acid bacteria have a good inhibitory effect on some spoilage bacteria and low-temperature bacteria. Lactic acid bacteria produce organic acids such as lactic acid and hydrogen peroxide. Among them, organic acids such as lactic acid can significantly reduce the environmental pH value and Eh (redox potential) value, making the intestine in an acidic environment, and have an antagonistic effect on pathogenic bacteria such as Shigella dysenteriae, Salmonella typhi, Salmonella paratyphi, Campylobacter, Staphylococcus, etc.; hydrogen peroxide can activate the catalase-thiocyanate system in milk, inhibit and kill Gram-negative bacteria, catalase-positive bacteria such as Pseudomonas, Escherichia coli and Salmonella. Therefore, lactic acid bacteria can be used to prevent and treat diarrhea, dysentery, enteritis, constipation and various diseases caused by intestinal dysfunction and skin inflammation.

[0004] Lactic acid bacteria products are mainly made by low temperature drying after liquid fermentation, which has high production cost and limits their use. Solid-state fermentation can also be used to produce lactic acid bacteria, but the general fermentation level is not as good as that of liquid. Wheat bran is a general term for the residue after wheat embryo and endosperm are extracted. Wheat bran is mainly composed of two parts, cortex and endosperm, accounting for about 15% of wheat grains. The content of protein, minerals, etc. in bran is richer than that in endosperm. Wheat bran is rich in dietary fiber, vitamin B, folic acid, and polyphenols, and is a cheap and high-quality culture medium for microorganisms. By optimizing nutrition, using bran as raw material and adding other auxiliary materials, a lactic acid bacteria culture with a high bacterial count can be obtained by solid-state fermentation. However, the bacterial count content of the product prepared by the existing fermentation process is not high, which seriously hinders the efficient application of bran. Summary of the invention

[0005] In view of the above technical problems, the present invention proposes a method for processing wheat bran fermented with high lactic acid bacteria count. The method can realize the processing value-added of wheat bran at low cost, improve the nutritional value, digestibility and antioxidant capacity of wheat bran, reduce anti-nutritional factors, and increase appetizing properties and probiotic effects.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] One of the purposes of the present invention is to provide a method for processing bran fermented with high lactic acid bacteria count, comprising the following steps: using bran as a substrate, amino acids, yeast extract powder, complex enzyme preparations, mineral ions and calcium carbonate as auxiliary materials, inoculating bacteria, performing solid-state anaerobic fermentation, and then low-temperature drying to generate a product.

[0008] The present invention increases suitable auxiliary materials and optimizes the fermentation process, so that after the bran is fermented, the cross-linked structure of lignocellulose is broken, and the bioactive substances are "released", so that phenolic acid is freed, and the contents of total phenol and ferulic acid are increased, thereby improving the antioxidant capacity of the fermented bran. The fermentation of bran by lactic acid bacteria can also produce organic acids such as lactic acid and flavor amino acids, so that the fermented bran has a special sour fragrance, which can stimulate the appetite of animals, promote animal feeding, and increase weight gain. In addition, the use of lactic acid bacteria fermentation can also reduce the content of anti-nutritional factors such as phytic acid in the bran, increase the content of oligosaccharides, and improve animal utilization. The most critical thing is that the number of bacteria in the bran can be increased.

[0009] Furthermore, the bran is fresh bran produced within half a month, and the dry basis ratio of the bran to the material (the material refers to the total amount of substrate + auxiliary materials) is greater than 93%.

[0010] Further, the amino acids consist of five crystalline amino acids: lysine, tryptophan, arginine, valine and glycine;

[0011] Based on the bran, the added amounts of amino acids are: 0.1-0.2% lysine, 0.05-0.1% tryptophan, 0.1-0.2% arginine, 0.01-0.02% valine and 0.03-0.05% glycine.

[0012] Furthermore, based on the bran, the yeast extract powder is added in an amount of 1.0-3.0%.

[0013] Furthermore, the composite enzyme preparation consists of xylanase, neutral protease, acid protease, alkaline protease, phytase, saccharifying enzyme, α-amylase and pullulanase;

[0014] Based on the bran, the added amount of the complex enzyme preparation is: xylanase 200-400U / g, neutral protease 100-300U / g, acid protease 100-300U / g, alkaline protease 200-400U / g, phytase 10-30U / g, saccharifying enzyme 100-200U / g, α-amylase 5-15U / g and pullulanase 2-4U / g.

[0015] Furthermore, the mineral ions are composed of magnesium ions, ferrous ions, manganese ions and zinc ions; the magnesium ions can be selected from magnesium sulfate; the ferrous ions can be selected from ferrous sulfate; the manganese ions can be selected from manganese sulfate; and the zinc ions can be selected from zinc sulfate.

[0016] Based on the bran, the added amounts of the mineral ions are: 0.05-0.1% of ferrous ions, 0.02-0.05% of magnesium ions, 0.01-0.02% of manganese ions and 0.01-0.02% of zinc ions.

[0017] Furthermore, based on the bran, the calcium carbonate addition amount is 4-5%.

[0018] Furthermore, the bacterial species is Pediococcus acidilactici, and the inoculation amount is 0.1-100 million CFU / g.

[0019] Furthermore, the conditions of the solid-state anaerobic fermentation are: fermentation temperature 40-45° C., initial material moisture 45-50%, and anaerobic fermentation time 40-48 hours.

[0020] Furthermore, the temperature of the low-temperature drying is less than 45° C., and the moisture content of the material after drying is less than 12%.

[0021] The second object of the present invention is to provide a fermented bran with a high lactic acid bacteria count prepared by the above processing method.

[0022] The third object of the present invention is to provide an application of the high lactic acid bacteria count fermented bran in the field of feed.

[0023] Compared with the prior art, the present invention has the following advantages and technical effects:

[0024] The present invention utilizes microbial fermentation technology, and on the basis of fully exploring the potential nutritional value of bran, further enhances the functional value, namely, the health-care effect of lactic acid bacteria and metabolites on the intestinal tract of animals. Through the optimization of the culture medium and the control of fermentation conditions, the lactic acid bacteria content reaches 60 billion CFU / g (dry basis), and a large number of beneficial metabolites are produced. The high lactic acid bacteria fermented bran produced by this process is equivalent to the lactic acid bacteria culture with bran as the substrate, which is not only an excellent feed raw material, but also a powerful animal "health product". The product is widely applicable to various farmed animals, and has the effects of improving the intestinal flora structure of animals, reducing intestinal diseases such as diarrhea, enhancing the body's antioxidant capacity, and improving the quality of livestock products. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The accompanying drawings constituting a part of the present invention are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the accompanying drawings:

[0026] Figure 1 The effect of fermentation time on the fermentation value of Pediococcus acidilactici in Example 1;

[0027] Figure 2 The effect of fermentation temperature on the fermentation value of Pediococcus acidilactici in Example 1;

[0028] Figure 3 The effect of the inoculum size on the fermentation value of Pediococcus acidilactici in Example 1;

[0029] Figure 4 The effect of the moisture content of the initial fermentation material on the fermentation value of Pediococcus acidilactici in Example 1;

[0030] Figure 5 It is a process flow chart of the present invention. DETAILED DESCRIPTION

[0031] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0032] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0033] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0034] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0035] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0036] The invention discloses a processing method of high lactic acid bacteria fermented bran, comprising the following steps: using bran as a substrate, amino acids (lysine, tryptophan, arginine, valine and glycine), yeast extract powder, compound enzyme preparation (xylanase, neutral protease, acid protease, alkaline protease, phytase, saccharifying enzyme, alpha-amylase and pullulanase), mineral ions (magnesium ions, ferrous ions, manganese ions and zinc ions) and calcium carbonate as auxiliary materials, inoculating bacteria, performing solid-state anaerobic fermentation, and then performing low-temperature drying to generate a product.

[0037] In the following examples of the present invention, the bran is fresh bran produced within half a month, accounting for more than 93% of the dry basis of the material. The bran sterilization method is conventional high-pressure sterilization in the art, and the specific sterilization method is not discussed as the creative method of the present invention, and the detailed method is not repeated.

[0038] In the following examples of the present invention, based on the substrate, the amount of amino acids added is: lysine 0.1-0.2%, tryptophan 0.05-0.1%, arginine 0.1-0.2%, valine 0.01-0.02% and glycine 0.03-0.05%. For example, the amount of lysine added is 0.1-0.2%, that is, the amount of lysine added is 0.1-0.2% of the substrate mass, the same below.

[0039] Illustratively, in the following embodiments of the present invention, the material contains 0.1%, 0.15%, 0.2% or any range or sub-range between the aforementioned ratios of lysine;

[0040] Illustratively, in the following embodiments of the present invention, the material contains tryptophan in an amount of 0.08%, 0.05%, 0.1% or any range or sub-range between the aforementioned ratios;

[0041] Illustratively, in the following embodiments of the present invention, the material contains 0.1%, 0.15%, 0.2% or any range or sub-range between the aforementioned ratios of arginine;

[0042] Illustratively, in the following embodiments of the present invention, the material contains valine in an amount of 0.01%, 0.02% or any range or sub-range between the aforementioned ratios;

[0043] Illustratively, in the following examples of the present invention, the material contains 0.03%, 0.04%, 0.05% or any range or sub-range between the aforementioned ratios of glycine.

[0044] In the following examples of the present invention, the yeast extract powder is added in an amount of 1.0-3.0% based on the substrate. For example, in the following examples of the present invention, the material contains 1%, 2%, 3% or any range or sub-range between the above ratios of yeast extract powder.

[0045] In the following embodiments of the present invention, based on the substrate, the added amount of the complex enzyme preparation is: xylanase 200-400U / g, neutral protease 100-300U / g, acid protease 100-300U / g, alkaline protease 200-400U / g, phytase 10-30U / g, saccharifying enzyme 100-200U / g, α-amylase 5-15U / g and pullulanase 2-4U / g.

[0046] Illustratively, in the following embodiments of the present invention, the material contains 200 U / g, 300 U / g, 400 U / g or any range or sub-range between the aforementioned ratios of xylanase;

[0047] Exemplarily, in the following embodiments of the present invention, the material contains 100U / g, 200U / g, 300U / g or any range or sub-range between the aforementioned ratios of neutral protease;

[0048] Illustratively, in the following embodiments of the present invention, the material contains 100U / g, 200U / g, 300U / g or any range or sub-range between the aforementioned ratios of acid protease;

[0049] Illustratively, in the following embodiments of the present invention, the material contains 200 U / g, 300 U / g, 400 U / g or any range or sub-range between the aforementioned ratios of alkaline protease;

[0050] Illustratively, in the following embodiments of the present invention, the material contains phytase in an amount of 10 U / g, 20 U / g, 30 U / g or any range or sub-range between the aforementioned ratios;

[0051] Illustratively, in the following embodiments of the present invention, the material contains saccharifying enzyme in an amount of 100 U / g, 150 U / g, 200 U / g or any range or sub-range between the aforementioned ratios;

[0052] Illustratively, in the following embodiments of the present invention, the material contains 5U / g, 10U / g, 15U / g or any range or sub-range between the aforementioned ratios of α-amylase;

[0053] Illustratively, in the following examples of the present invention, the material contains 2 U / g, 3 U / g, 4 U / g or any range or sub-range between the aforementioned ratios of pullulanase.

[0054] In the following examples of the present invention, based on the substrate, the added amounts of the mineral ions are: 0.05-0.1% ferrous ions, 0.02-0.05% magnesium ions, 0.01-0.02% manganese ions and 0.01-0.02% zinc ions.

[0055] Illustratively, in the following embodiments of the present invention, the material contains 0.05%, 0.08%, 0.1% or any range or sub-range between the aforementioned ratios of ferrous ions;

[0056] Illustratively, in the following embodiments of the present invention, the material contains 0.02%, 0.04%, 0.05% or any range or sub-range between the aforementioned ratios of magnesium ions;

[0057] Illustratively, in the following embodiments of the present invention, the material contains 0.01%, 0.02% or any range or sub-range between the aforementioned ratios of manganese ions;

[0058] Illustratively, in the following examples of the present invention, the material contains 0.01%, 0.02% or any range or sub-range between the aforementioned ratios of zinc ions.

[0059] In the following examples of the present invention, the amount of calcium carbonate added is 4-5% based on the substrate. The material contains 4%, 4.5%, 5% or any range or sub-range between the above ratios of calcium carbonate.

[0060] In the following embodiments of the present invention, the bacterial species is Pediococcus acidilactici, preferably Pediococcus acidilactici CICC 24367, which is purchased from the market; the inoculation amount is 100-100 million CFU / g. Exemplarily, in the following embodiments of the present invention, the bacterial species inoculation amount can be selected from 100 million CFU / g, 250 million CFU / g, 500 million CFU / g, and 100 million CFU / g.

[0061] In the following examples of the present invention, the conditions of the solid-state anaerobic fermentation are: fermentation temperature 40-45° C., material moisture 45-50%, and anaerobic fermentation time 40-48 h.

[0062] In the following examples of the present invention, the temperature of the low-temperature drying is less than 45° C., and the moisture content of the material after drying is less than 12%.

[0063] The indicators of the product obtained by the above processing method are: lactic acid bacteria count 60 billion / g (dry basis), crude protein content>16%, acid-soluble protein content>10%, and calcium lactate>8%.

[0064] The raw materials used in the present invention are all purchased from the market.

[0065] The technical solution of the present invention is further described below through examples, and the actual application effect of the fermented bran prepared by the present invention can be reflected through the following test data. Figure 5 It is a process flow chart of the present invention.

[0066] Example 1

[0067] Optimization of Fermentation Conditions of Pediococcus acidilactici

[0068] A single factor experiment was set up: inoculation amount, fermentation temperature, material moisture and fermentation time were used as single factor variables to examine their effects on the fermentation degree of Pediococcus acidilactici in fermented bran.

[0069] Select 2 kg of sterilized fresh wheat bran without impurities, mold, oxidation or deterioration, and then weigh 2% yeast extract powder (i.e., based on 100% dry basis weight of wheat bran, the addition amount of yeast extract powder is 2%, the same below), 4.5% calcium carbonate, 0.08% ferrous sulfate (calculated as iron ions, the same below), 0.04% magnesium sulfate (calculated as magnesium ions, the same below), 0.02% manganese sulfate (calculated as manganese ions, the same below), and 0.06% zinc sulfate (calculated as zinc ions, the same below). .02%, lysine 0.15%, tryptophan 0.08%, arginine 0.15%, valine 0.02%, glycine 0.04%, xylanase 300U / g, neutral protease 200U / g, acid protease 200U / g, alkaline protease 300U / g, phytase 20U / g, saccharifying enzyme 150U / g, α-amylase 10U / g, pullulanase 3U / g, and mix evenly with the bran to obtain a mixed material.

[0070] There were 25 treatment groups in the experiment. 100 g of the above mixed materials were taken from each group, and solid-state anaerobic fermentation was carried out after inoculation with Pediococcus acidilactici. The specific settings were as follows:

[0071] 1) The fermentation temperature was fixed at 42°C, the inoculation amount was 50 million CFU / g, the moisture content of the material was adjusted to 45%, and the fermentation was performed for 0h, 8h, 16h, 24h, 32h, 40h, 48h, and 56h respectively;

[0072] 2) The fermentation time was fixed at 48 h, the inoculation amount was 50 million CFU / g, the moisture content of the material was adjusted to 45%, and the fermentation temperature was controlled at 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, and 50°C respectively;

[0073] 3) The fermentation time was fixed at 48 h, the fermentation temperature was 42 ° C, the material moisture was adjusted to 45%, and the inoculation amount was controlled to 50 million CFU / g, 100 million CFU / g, 250 million CFU / g, 500 million CFU / g, and 100 million CFU / g respectively;

[0074] 4) The fermentation time was fixed at 48 h, the fermentation temperature was 42° C., the inoculation amount was 50 million CFU / g, and the moisture content of the initial fermentation material was controlled at 35%, 40%, 45%, 50%, and 55% respectively.

[0075] The above four groups of experiments were conducted on data from different batches. Therefore, there are certain errors in the fermentation products under the same conditions in different batches, which are within a reasonable range.

[0076] After fermentation, the number of lactic acid Pediococcus in each of the above treatments was determined.

[0077] Figure 1 The effect of fermentation time on the fermentation value of Pediococcus acidilactici in Example 1. The results show that when Pediococcus acidilactici is cultured using wheat bran as a substrate, the fermentation time exceeds 40 hours, and the bacterial count reaches more than 60 billion CFU / g (wet basis). Considering the comprehensive production cost, the fermentation time is preferably between 40-48 hours.

[0078] Figure 2 The effect of fermentation temperature on the fermentation value of Pediococcus acidilactici in Example 1. The results show that when wheat bran is used as a substrate to culture Pediococcus acidilactici, the fermentation temperature of Pediococcus acidilactici can reach more than 60 billion CFU / g (wet basis) when the fermentation temperature is between 40-45°C.

[0079] Figure 3 The effect of inoculation amount on the fermentation value of Pediococcus acidilactici in Example 1. The results show that when wheat bran is used as a substrate to culture Pediococcus acidilactici, the inoculation amount of the strain is between 0.1-100 million CFU / g, the Pediococcus acidilactici can reach more than 60 billion CFU / g (wet basis).

[0080] Figure 4 The effect of moisture content of the initial fermentation material on the fermentation value of Pediococcus acidilactici in Example 1. The results show that when wheat bran is used as a substrate to culture Pediococcus acidilactici, the Pediococcus acidilactici can reach more than 60 billion CFU / g (wet basis) when the moisture content of the material is between 45-50%.

[0081] In summary, during the fermentation process, the fermentation temperature of the material is controlled between 40-45°C, the moisture content of the material is controlled at 45-50%, and after 40-48 hours of anaerobic fermentation under precise temperature and humidity control conditions, the material is dried at a low temperature below 45°C to a moisture content of less than 12%. The lactic acid bacteria content of the prepared high lactic acid bacteria count fermented bran reaches more than 60 billion CFU / g (wet basis).

[0082] Example 2

[0083] Small-scale experiment on fermentation of bran with high lactic acid bacteria count

[0084] Select 10 kg of sterilized fresh wheat bran without impurities, mold, oxidation or deterioration; then weigh 1% yeast extract, 4% calcium carbonate, 0.05% ferrous sulfate, 0.02% magnesium sulfate, 0.01% manganese sulfate, 0.01% zinc sulfate, 0.1% lysine, 0.05% tryptophan, 0.1% arginine, 0.01% valine, 0.03% glycine, 200 U / g xylanase, 10% neutral protease; 0U / g, acid protease 100U / g, alkaline protease 200U / g, phytase 10U / g, saccharifying enzyme 100U / g, α-amylase 5U / g, pullulanase 2U / g, 100 million CFU / g of Pediococcus acidilactici are added, after mixed inoculation, the fermentation temperature is controlled between 40-45°C, the material moisture is 46%, anaerobic fermentation is carried out for 48h, and then low-temperature drying below 45°C is carried out to prepare fermented bran with high lactic acid bacteria count.

[0085] After preparing the fermented bran with high lactic acid bacteria count according to the fermentation method of this embodiment, the crude protein, acid-soluble protein, ferulic acid, reducing sugar, neutral detergent fiber, acid detergent fiber, calcium lactate content, lactic acid bacteria count and pH of the wheat bran raw material and the fermented bran with high lactic acid bacteria count were measured, and the results are shown in Table 1.

[0086] Table 1 Determination of the indexes of bran and raw materials fermented with high lactic acid bacteria count in the small test

[0087]

[0088] The results showed that in the high lactic acid bacteria count fermented bran prepared by the fermentation method of Example 2, the lactic acid bacteria content reached 61.7 billion CFU / g (dry basis), the crude protein content reached 18.72%, and the acid-soluble protein content reached 10.82%.

[0089] Example 3

[0090] Pilot test of fermentation of bran with high lactic acid bacteria count

[0091] 1000 kg of fresh wheat bran without impurities, mold, oxidation or deterioration was sterilized and then 3% yeast extract, 5% calcium carbonate, 0.1% ferrous sulfate, 0.05% magnesium sulfate, 0.02% manganese sulfate, 0.02% zinc sulfate, 0.2% lysine, 0.1% tryptophan, 0.2% arginine, 0.02% valine, 0.05% glycine, 400 U / g xylanase, 3% neutral protease were weighed. 00U / g, acid protease 300U / g, alkaline protease 400U / g, phytase 30U / g, saccharifying enzyme 200U / g, α-amylase 15U / g, pullulanase 4U / g, 100 million CFU / g of Pediococcus acidilactici are added, after mixed inoculation, the fermentation temperature is controlled between 40-45°C, the material moisture is 48%, anaerobic fermentation is carried out for 48h, and then low-temperature drying below 45°C is carried out to prepare fermented bran with high lactic acid bacteria count.

[0092] After preparing the fermented bran with high lactic acid bacteria count according to the fermentation method of this embodiment, the crude protein, acid-soluble protein, ferulic acid, reducing sugar, neutral detergent fiber, acid detergent fiber, calcium lactate content, lactic acid bacteria count and pH of the wheat bran raw material and the fermented bran with high lactic acid bacteria count were measured. The results are shown in Table 2.

[0093] Table 2 Determination of the parameters of the pilot high lactic acid bacteria fermented bran and raw materials

[0094]

[0095] The results showed that the fermented bran with high lactic acid bacteria count prepared by the fermentation method of Example 3 had a lactic acid bacteria content of 64 billion CFU / g (dry basis), a crude protein content of 18.81%, and an acid-soluble protein content of 11.25%.

[0096] Example 4

[0097] Effects of fermented bran with high lactic acid bacteria count on production performance and egg quality of laying hens

[0098] 1. Experimental design

[0099] 600 healthy laying hens aged 30 weeks were selected and randomly divided into 5 groups (1 control group and 4 experimental groups), with 4 replicates in each group and 30 laying hens in each replicate. The laying hens in each replicate were evenly distributed to the upper, middle and lower layers of the same ladder-type laying hens cage, with 10 laying hens on each layer. The pre-feeding period was 7 days and the experimental period was 30 days.

[0100] Control group: complete feed, using corn-soybean meal basic feed formula;

[0101] Experimental group: 1%, 3%, 5% and 7% of the fermented bran with high lactic acid bacteria count produced in Example 3 were added to the complete formula feed respectively.

[0102] The feed formula used in the experiment is shown in Table 3.

[0103] Table 3 Composition and nutritional level of experimental diets

[0104]

[0105] 2. Index determination and methods

[0106] 2.1 Determination of production performance of laying hens

[0107] Eggs were collected twice a day, and the number of eggs laid, total egg mass, number of broken eggs, and feed intake were recorded for each replicate. The egg production rate, feed-egg ratio, average egg mass, average daily feed intake, and egg breaking rate were calculated.

[0108] 2.2 Egg quality determination

[0109] On the day when the experiment ended, 10 eggs were randomly selected from each replicate, with a total of 40 eggs in each treatment, and the egg quality was measured on the day of the experiment. Egg quality, albumen height, yolk color, and Haugh units were measured using an egg quality tester; yolk quality was measured using an electronic balance; the longitudinal diameter, transverse diameter, yolk height, and yolk diameter of the laying hen were measured using a vernier caliper, and the thickness of the sharp end, middle end, and blunt end of the eggshell was measured after removing the eggshell membrane; and the eggshell strength was measured using an eggshell strength tester.

[0110] Yolk index = yolk height / yolk diameter

[0111] Yolk ratio = yolk mass / egg mass × 100%

[0112] Egg-shaped index = vertical diameter / horizontal diameter

[0113] 3. Data processing

[0114] The data were processed using Microsoft Excel 2010 and analyzed using IBM SPSS Statistics 27 software. Duncan's method was used for multiple comparisons, and P < 0.05 indicated a significant difference.

[0115] 4. Results and Analysis

[0116] 4.1 Effects of high lactic acid bacteria count fermented bran on laying hen production performance

[0117] The results of the effect of high lactic acid bacteria count fermented bran on the production performance of laying hens are shown in Table 4.

[0118] Table 4 Effects of high lactic acid bacteria count fermented bran on laying hen production performance

[0119]

[0120] Note: Different letters indicate significant differences between data (P<0.05), the same as in the following table.

[0121] According to Table 4, there were no significant differences in the average daily feed intake, average egg weight, and feed-egg ratio among the five treatment groups; the egg production rate of the experimental group with 5% and 7% addition of high lactic acid bacteria fermented bran was significantly higher than that of the control group; the egg breaking rate of the experimental group with 3%, 5%, and 7% addition of high lactic acid bacteria fermented bran was significantly lower than that of the control group, and the formula cost was lower than that of the control group, with the highest formula cost difference of 30 yuan / ton. The results show that the use of 3%-7% high lactic acid bacteria fermented bran in laying hen feed can significantly increase its egg production rate, reduce the egg breaking rate, and reduce feed costs. That is, the use of high lactic acid bacteria fermented bran can significantly improve the production performance of laying hens.

[0122] 4.2 Effect of high lactic acid bacteria count fermented bran on egg quality

[0123] The results of the effect of high lactic acid bacteria fermented bran on egg quality are shown in Table 5. The Haugh units of the experimental groups with 5% and 7% high lactic acid bacteria fermented bran addition were significantly higher than those of the control group; there were no significant differences in the other indicators among the treatment groups. The results show that the use of 5%-7% high lactic acid bacteria fermented bran in laying hen feed can significantly increase the Haugh units of eggs, that is, the use of high lactic acid bacteria fermented bran can increase the protein consistency and improve the quality of eggs.

[0124] Table 5 Effect of high lactic acid bacteria count fermented bran on egg quality

[0125]

[0126] Example 5

[0127] Effects of high lactic acid bacteria count fermented bran on reproductive performance and serum biochemical and antioxidant indexes in pregnant sows

[0128] 1. Experimental design

[0129] Sixty first-parity pregnant sows with similar body condition and the same expected delivery date were selected and randomly divided into 6 treatment groups, with 10 pregnant sows in each treatment group, and each pregnant sow was a biological replicate. The experiment started at 60 days of pregnancy and ended after delivery.

[0130] Control group: complete feed, with corn-soybean meal as the main protein raw materials;

[0131] Experimental group: 3%, 6%, 9%, 12% and 15% of the fermented bran with high lactic acid bacteria count produced in Example 3 were added to the complete feed respectively.

[0132] Feed formula and ingredients

[0133] The feed formula is shown in Table 6.

[0134] Table 6 Composition and nutritional components of experimental diets

[0135]

[0136] 3. Measurement indicators and methods

[0137] 3.1 Reproductive performance

[0138] Within 12 hours after farrowing, the number of piglets born and the number of live piglets of each sow were recorded, and the litter weight at birth was measured.

[0139] 3.2 Serum sample collection and index determination

[0140] On the 110th day of pregnancy, 5 sows were randomly selected from each group before morning feeding, and 10 mL of blood was collected through the anterior vena cava. The blood was allowed to stand for 30 minutes and centrifuged at 3000 r / min for 10 minutes to precipitate serum. The supernatant was collected in a centrifuge tube and stored in a -20°C refrigerator for the determination of serum biochemical indices and antioxidant indices.

[0141] The contents of total protein (TP), albumin (ALB), globulin (GLB), total cholesterol (TC), TG, urea nitrogen (BUN), glucose (GLU) and the activities of alanine aminotransferase (AST) and aspartate aminotransferase (ALT) in serum were determined by automatic biochemical analyzer.

[0142] Kits were used to determine the contents of superoxide dismutase (SOD), total antioxidant capacity (T-AOC), catalase (CAT), malondialdehyde (MDA), and glutathione peroxidase (GSH-Px).

[0143] 3.3 Data processing

[0144] Statistical analysis and variance analysis of each element index were performed using Microsoft EXCEL 2010 software and IBM SPSS Statistics 27 software.

[0145] 4. Results and Analysis

[0146] 4.1 Effects of high lactic acid bacteria count fermented bran on reproductive performance of pregnant sows

[0147] The results of the effect of high lactic acid bacteria fermented bran on the reproductive performance of pregnant sows are shown in Table 7. The litter weight and average weight of piglets at birth in the five experimental groups were significantly higher than those in the control group, and the average weight of piglets at birth increased by up to 17.42%. The formula costs of the five experimental groups were lower than those of the control group, and the highest formula cost difference reached 81 yuan / ton; the litter weight and average weight of piglets at birth in the experimental groups with 6%, 9%, and 12% addition of high lactic acid bacteria fermented bran were significantly higher than those in other treatment groups. The results show that the use of 3%-15% high lactic acid bacteria fermented bran in pregnant sow feed can significantly increase the litter weight and the weight of piglets at birth, and can significantly reduce feed costs; the addition of high lactic acid bacteria fermented bran at 6%-12% has the most outstanding effect; that is, the use of high lactic acid bacteria fermented bran can significantly improve the reproductive performance of sows.

[0148] Table 7 Effects of high lactic acid bacteria count fermented bran on reproductive performance of pregnant sows

[0149]

[0150]

[0151] 4.2 Effects of fermented bran with high lactic acid bacteria count on serum biochemical parameters of pregnant sows

[0152] The results of the effects of high lactic acid bacteria count fermented bran on serum biochemical parameters of pregnant sows are shown in Table 8.

[0153] Table 8 Effects of fermented bran with high lactic acid bacteria count on serum biochemical parameters of pregnant sows

[0154]

[0155] According to Table 8, the globulin content of the experimental groups with high lactic acid bacteria count fermented bran addition of 6%, 9%, 12%, and 15% was significantly increased compared with the control group, and the white / globulin ratio was significantly decreased; the total cholesterol, urea nitrogen, alanine aminotransferase and alanine aminotransferase contents of the experimental groups with high lactic acid bacteria count fermented bran addition of 9%, 12%, and 15% were significantly decreased compared with the control group; the results showed that adding high lactic acid bacteria count fermented bran can improve the serum biochemical indicators of pregnant sows, that is, feeding high lactic acid bacteria count fermented bran can improve the liver function, kidney function and immune ability of pregnant sows, and the effect of improving the serum biochemical indicators of pregnant sows is best when the addition of high lactic acid bacteria count fermented bran is between 9% and 15%.

[0156] 4.3 Effects of fermented bran with high lactic acid bacteria count on serum antioxidant indexes in pregnant sows

[0157] The results of the effects of high lactic acid bacteria count fermented bran on serum antioxidant indicators of pregnant sows are shown in Table 9.

[0158] Table 9 Effects of fermented bran with high lactic acid bacteria count on serum antioxidant indexes in pregnant sows

[0159]

[0160] According to Table 9, the total antioxidant capacity of the test groups with 6%, 9%, 12%, and 15% of high lactic acid bacteria fermented bran was significantly higher than that of the control group, and the content of antioxidant dismutase was also significantly higher than that of the control group; the blood malondialdehyde content of the test groups with 9%, 12%, and 15% of high lactic acid bacteria fermented bran was significantly lower than that of the control group. The results show that adding high lactic acid bacteria fermented bran can improve the antioxidant capacity of pregnant sows and effectively reduce the formation of lipid peroxides in the body. The antioxidant capacity of pregnant sows is particularly obvious when the addition of high lactic acid bacteria fermented bran is between 6% and 15%.

[0161] In summary, fermentation of bran with high lactic acid bacteria counts produces organic acids such as lactic acid and flavor amino acids, giving it a special sour aroma that can stimulate animal appetite, promote animal feeding, and increase weight gain. Lactic acid bacteria fermentation reduces the content of anti-nutritional factors such as phytic acid in bran, increases the content of oligosaccharides, improves animal utilization, increases antioxidant capacity and immunity, and improves the quality of livestock products. This product is an excellent functional feed raw material, widely used in egg-laying poultry, meat poultry, sows, aquatic products, ruminants and economic animals.

[0162] The above are only preferred specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A method for processing bran with high lactic acid bacteria count, characterized in that: The following steps are involved: With bran as substrate, amino acids, yeast extract powder, complex enzyme preparations, mineral ions and inorganic salts as auxiliary materials, solid-state anaerobic fermentation is carried out after inoculation of bacteria, and then the product is produced through low-temperature drying.

2. The high lactic acid bacteria count fermented bran processing method according to claim 1, characterized in that: The bran is fresh bran, accounting for more than 90% of the dry basis of the fermented material.

3. The high lactic acid bacteria count fermented bran processing method according to claim 1, characterized in that: The amino acids are composed of five crystalline amino acids: lysine, tryptophan, arginine, valine and glycine; Based on the mass of the bran, the added amounts of the amino acids are: 0.1-0.2% lysine, 0.05-0.1% tryptophan, 0.1-0.2% arginine, 0.01-0.02% valine and 0.03-0.05% glycine.

4. The method for processing bran with high lactic acid bacteria count according to claim 1, characterized in that: Based on the mass of the bran, the added amount of the yeast extract powder is 1.0-3.0%.

5. The method for processing bran with high lactic acid bacteria count according to claim 1, characterized in that: The composite enzyme preparation consists of xylanase, neutral protease, acid protease, alkaline protease, phytase, saccharifying enzyme, α-amylase and pullulanase; Based on the mass of the bran, the added amount of the complex enzyme preparation is: xylanase 200-400U / g, neutral protease 100-300U / g, acid protease 100-300U / g, alkaline protease 200-400U / g, phytase 10-30U / g, saccharifying enzyme 100-200U / g, α-amylase 5-15U / g and pullulanase 2-4U / g.

6. The method for processing bran with high lactic acid bacteria count according to claim 1, characterized in that: The mineral ions are composed of magnesium ions, ferrous ions, manganese ions and zinc ions; Based on the mass of the bran, the added amounts of the mineral ions are: 0.05-0.1% of ferrous ions, 0.02-0.05% of magnesium ions, 0.01-0.02% of manganese ions and 0.01-0.02% of zinc ions.

7. The method for processing bran with high lactic acid bacteria count according to claim 1, characterized in that: Based on the mass of the bran, the added amount of the inorganic salt is 4-5%; the inorganic salt is calcium carbonate.

8. The method for processing bran with high lactic acid bacteria count according to claim 1, characterized in that: The bacterial species is Pediococcus acidilactici, and the inoculation amount is 0.1-100 million CFU / g; and / or The conditions of the solid-state anaerobic fermentation are: fermentation temperature 40-45°C, initial material moisture 45-50%, anaerobic fermentation time 40-48h; and / or The temperature of the low-temperature drying is less than 45° C., and the moisture content of the material after drying is less than 12%.

9. A fermented bran with a high lactic acid bacteria count prepared by the method according to any one of claims 1 to 8.

10. Use of the fermented bran with high lactic acid bacteria count as claimed in claim 9 in the field of feed.

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