Method for producing cattle and sheep feed by taking pleurotus eryngii mushroom residues as raw materials through step-by-step fermentation

Abstract

The invention belongs to the technical field of edible fungus chaff feed, and particularly relates to a method for producing cattle and sheep feed through step-by-step fermentation by taking pleurotus eryngii chaff as a raw material, and the method comprises the following steps: S1, preparing a fungus chaff fermentation base material; s2, preparing a candida utilis seed solution and a lactobacillus plantarum seed solution; s3, inoculating the candida utilis seed solution into the mushroom bran fermentation base material for fermentation; and S4, inoculating the fermentation product with a lactobacillus plantarum seed solution, and fermenting again to obtain the step-by-step fermented mushroom bran feed. According to the method for producing the cattle and sheep feed through step-by-step fermentation by taking the pleurotus eryngii mushroom bran as the raw material, the nutritional level of the mushroom bran feed can be effectively improved, and the edible mushroom bran can be reasonably and fully utilized as an unconventional feed resource; the feed additive has important significance in solving the problem of feed shortage in the cattle and sheep breeding process, promoting healthy development of the breeding industry, saving resources and protecting the environment.

Description

Technical Field

[0001] The invention belongs to the technical field of edible mushroom bran feed, and particularly relates to a method for producing cattle and sheep feed by step-by-step fermentation using Pleurotus eryngii bran as a raw material. Background Art

[0002] At present, the main ways to utilize edible mushroom bran include cultivating edible mushrooms, substrate organic fertilizer, feed and active substance extraction. Pleurotus eryngii bran is the remaining part of the cultivation substrate after the fruiting body is harvested. It generally contains fibrous materials such as sawdust, crop straw, corn cobs, cottonseed hulls, and auxiliary materials such as rice bran, bran, and soybean meal. After harvesting, the bran still retains a large amount of mycelium and various metabolites, which are extremely nutritious. Reasonable and full use of edible mushroom bran as an unconventional feed resource can solve the problem of feed shortage in livestock and poultry breeding, which has a huge driving effect on the development of the breeding industry, while also saving resources and protecting the environment.

[0003] Mushroom bran is a good roughage resource, but direct feeding will result in low digestibility and requires further processing. A large number of studies have shown that microbial fermentation is an effective way to improve the nutrition of edible mushroom bran. Microorganisms produce organic acids and other nutrients during their growth and reproduction, which can improve palatability and digestibility, making it have higher nutritional and feeding value. At the same time, microbial fermentation also brings a large amount of bacterial protein to the feed.

[0004] Crude protein (CP) in feed mainly includes two parts of nitrogen-containing substances: true protein (TP) and non-protein. True protein TP is pure protein from which non-protein nitrogen-containing compounds have been removed. Its proportion in the total protein in feed has a particularly important impact on feed quality.

[0005] The inventors used utilis and Lactobacillus plantarum to treat Pleurotus eryngii bran in a step-by-step fermentation process, studied the changes in the contents of crude protein CP, crude fat EE, crude fiber CF, neutral detergent fiber NDF, acid detergent fiber ADF and acid detergent lignin ADL in Pleurotus eryngii bran after step-by-step fermentation, and established a process for improving the nutritional level of Pleurotus eryngii bran by microbial step-by-step fermentation, which is of great significance for the high-value utilization of edible mushroom bran.

[0006] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention and should not be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art. Summary of the invention

[0007] The purpose of the present invention is to provide a method for producing cattle and sheep feed by step-by-step fermentation using Pleurotus eryngii husks as raw materials.

[0008] To achieve the above object, the present invention provides the following technical solutions:

[0009] A method for producing cattle and sheep feed by fermenting Pleurotus eryngii husks as raw materials in steps, comprising the following steps:

[0010] S1. Prepare the fermentation base of fungus husk

[0011] S2. Preparation of seed solution

[0012] Candida utilis was inoculated into YPD medium and cultured in a constant temperature shaker at 30°C and 150 r / min for 24 h. After two generations of continuous culture, the concentration was adjusted to 1×10 8 cfu / mL, and the Candida utilis seed solution was obtained;

[0013] Lactobacillus plantarum was inoculated into MRS medium and cultured at 37°C for 24 h. After two generations of continuous culture, the concentration was adjusted to 1×10 8 cfu / mL, and Lactobacillus plantarum seed solution was obtained;

[0014] S3. First stage fermentation

[0015] The Candida utilis seed liquid was inoculated into the fermentation base material of the fungus bran at an inoculation rate of 6%, and the fermentation was carried out in a constant temperature incubator at 30°C for 60 hours with an oxygen flow rate of 8L / min and the material was turned once every 12 hours;

[0016] S4. Second stage fermentation

[0017] The Lactobacillus plantarum seed liquid was inoculated into the first stage fermentation product at an inoculum rate of 4%, mixed evenly and put into a respiratory fermentation bag, compacted to remove the air in the bag and sealed, and fermented at a constant temperature of 37° C. for 8 days to obtain the step-fermented bran feed.

[0018] Preferably, in S1, the preparation method of the fermented mushroom bran base material is: evenly mix Pleurotus eryngii mushroom bran and molasses water at a material-liquid ratio of 10g:1mL, and sterilize at 121°C for 40min; wherein the base material of the Pleurotus eryngii mushroom bran is: bagasse, corn cobs, corn flour, bran and light calcium.

[0019] Preferably, the brix of the molasses water is 16° Bx.

[0020] Preferably, in S2, the formula of YPD medium is: 10.0 g yeast powder, 20.0 g glucose, 20.0 g peptone, 1000 mL distilled water, pH 6.0±0.2, sterilized at 121°C for 15 min.

[0021] Preferably, in S2, the formula of MRS medium is: casein digest 10.0 g, beef extract powder 10.0 g, yeast extract powder 4.0 g, triammonium citrate 2.0 g, sodium acetate 5.0 g, MgSO4·7H2O 0.2 g, MnSO4·4H2O0.05 g, dimethyl phosphate 2.0 g, glucose 20.0 g, Tween-80 1.0 g, distilled water 1000 mL, pH 5.7±0.2, sterilized at 121°C for 15 min.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] The present invention uses Candida utilis and Lactobacillus plantarum to ferment the shiitake mushroom bran in steps, which can significantly increase the crude protein and crude fat content of the shiitake mushroom bran, and has a significant effect on reducing the crude fiber, neutral detergent fiber, acid detergent fiber and acid detergent lignin content, especially the crude protein content of the bran after step-by-step fermentation reaches 18.80%, which is 83.25% higher than the 10.26% of the original bran. It can be seen that the method of the present invention of producing cattle and sheep feed by step-by-step fermentation using the shiitake mushroom bran as raw material can effectively improve the nutritional level of the bran feed, is conducive to the rational and full use of edible bran as an unconventional feed resource, and is of great significance for solving the problem of feed shortage in cattle and sheep breeding, promoting the healthy development of the breeding industry, saving resources, and protecting the environment. DETAILED DESCRIPTION

[0024] The following is a clear and complete description of the technical solution of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by technicians in this field without creative work are within the scope of protection of the present invention.

[0025] Materials and Methods

[0026] 1. Main Materials and Reagents

[0027] Pleurotus eryngii bran: provided by the edible fungus factory of the Institute of Microbiology, Guangxi Academy of Agricultural Sciences. Its base materials are sugarcane bagasse, corn cobs, corn flour, bran and light calcium.

[0028] Molasses: purchased from Mingyang Sugar Factory of Nanning Sugar Company, with an initial Brix of 78°Bx. It was diluted with water to a suitable Brix when used.

[0029] Candida utilis CICC32722 and Lactobacillus plantarum CICC22703 were purchased from China Industrial Culture Collection Center (CICC).

[0030] YPD medium: 10.0 g yeast powder, 20.0 g glucose, and 20.0 g peptone, dissolved in 1000 mL distilled water, pH 6.0±0.2, sterilized at 121°C for 15 min.

[0031] MRS medium: 10.0 g of casein digest, 10.0 g of beef extract powder, 4.0 g of yeast extract powder, 2.0 g of triammonium citrate, 5.0 g of sodium acetate, 0.2 g of MgSO4·7H2O, 0.05 g of MnSO4·4H2O, 2.0 g of dimethyl phosphate, 20.0 g of glucose, and 1.0 g of Tween-80, dissolved in 1000 mL of distilled water, pH 5.7±0.2, sterilized at 121°C for 15 min.

[0032] Mushroom bran fermentation medium: Add molasses water (brix of 16°Bx) to the mushroom bran at a solid-liquid ratio of 10g:1mL and mix well. Weigh 200g of the mixed material into a 250mL conical flask, cover the bottle mouth with a breathable rubber stopper, set the pH to natural, and sterilize at 121°C for 40min.

[0033] 2. Methods

[0034] 2.1 Preparation of seed solution

[0035] Candida utilis was inoculated into YPD medium and cultured in a constant temperature shaker at 30°C and 150 r / min for 24 h. After two generations of continuous culture, the concentration was adjusted to 1×10 8 CFU / mL, stored in a refrigerator at 4℃ for future use.

[0036] Lactobacillus plantarum was inoculated into MRS medium and cultured at 37°C for 24 h. After two generations of continuous culture, the concentration was adjusted to 1×10 8 CFU / mL, stored in a refrigerator at 4℃ for future use.

[0037] 2.2 Basic ingredient content of edible mushroom bran

[0038] The edible mushroom sticks were broken into bags and mixed evenly, and the weight of the samples was reduced to 250 g by quartering, dried at 65 °C to constant weight, crushed by a high-speed sample grinder and passed through a 40-mesh sieve. The crude protein, crude fat, crude fiber, neutral detergent fiber, acid detergent fiber, acid detergent lignin and crude ash contents of the mushroom bran were determined by sampling. Three parallel samples were used, and the average value of the parallel samples was taken as the CK control sample. The results are shown in Table 1.

[0039] 2.3 Step fermentation

[0040] The sterilized fermentation medium of the fungus bran was cooled to room temperature, and the Candida utilis seed liquid was inoculated into the fermentation base of the fungus bran at a 6% inoculation rate, and placed in a 30°C constant temperature incubator for fermentation for 60 hours, with an oxygen flow rate of 8L / min, and the material was turned once every 12 hours; after the fermentation, the Lactobacillus plantarum seed liquid was inoculated into the fermentation product at a 4% inoculation rate, mixed evenly, and put into a respiratory fermentation bag, compacted to remove the air in the bag and sealed, and fermented at a constant temperature of 37°C for 8 days to obtain the fungus bran feed. After opening the bag, the fungus bran feed was randomly sampled to determine the content of each nutrient component, and 3 parallel samples were used to take the average value of the parallel samples. The results are shown in Table 1.

[0041] Table 1 Changes in nutritional components of Pleurotus eryngii husk before and after step-by-step fermentation

[0042]

[0043] Note: Different lowercase letters in the same column indicate significant differences (P<0.05).

[0044] As shown in Table 1, the Pleurotus eryngii bran was fermented step by step with Candida utilis and Lactobacillus plantarum. After fermentation, the crude protein CP content of the Pleurotus eryngii bran reached 18.80%, which was 83.25% higher than that before fermentation, and the improvement effect was significant (P < 0.05); the crude fiber CF content of the Pleurotus eryngii bran after step-by-step fermentation was 21.21%, which was 38.16% lower than that before fermentation (34.30%), and the effect was significant (P < 0.05); the crude fat EE content of the Pleurotus eryngii bran after fermentation was 6.80%, which was 18.06% higher than that before fermentation, indicating that the step-by-step fermentation had a significant effect on improving its crude fat content (P < 0.05).

[0045] After step-by-step fermentation, the contents of neutral detergent fiber NDF, acid detergent fiber ADF and acid detergent lignin ADL in Pleurotus eryngii bran were 45.11%, 33.25% and 7.71%, respectively, which were 29.08%, 31.89% and 37.37% lower than those before fermentation, and the effects were significant (P < 0.05); the crude ash Ash content of Pleurotus eryngii bran after step-by-step fermentation was 9.75%, which was slightly higher than that before fermentation, but the difference was not significant (P > 0.05).

[0046] In summary, the present invention uses Candida utilis and Lactobacillus plantarum to ferment the shiitake mushroom bran in steps, which can significantly increase the crude protein and crude fat content of the shiitake mushroom bran, and has a significant effect on reducing the crude fiber, neutral detergent fiber, acid detergent fiber and acid detergent lignin content, especially the crude protein content of the bran after step-by-step fermentation reaches 18.80%, which is 83.25% higher than the 10.26% of the original bran. It can be seen that the method of producing cattle and sheep feed by step-by-step fermentation using shiitake mushroom bran as raw material can effectively improve the nutritional level of bran feed, is conducive to the rational and full use of edible bran as an unconventional feed resource, and is of great significance for solving the problem of feed shortage in cattle and sheep breeding, promoting the healthy development of breeding industry, saving resources and protecting the environment.

[0047] The foregoing description of specific exemplary embodiments of the present invention is for the purpose of illustration and demonstration. These descriptions are not intended to limit the present invention to the precise form disclosed, and it is clear that many changes and variations can be made based on the above teachings. The purpose of selecting and describing the exemplary embodiments is to explain the specific principles of the present invention and its practical application, so that those skilled in the art can realize and utilize various different exemplary embodiments of the present invention and various different selections and changes. The scope of the present invention is intended to be limited by the claims and their equivalents.

Claims

1. A method for producing cattle and sheep feed by fermenting Pleurotus eryngii husks as raw materials in steps, characterized in that: The following steps are involved: S1. Prepare the fermentation base of fungus husk S2. Preparation of seed solution Candida utilis was inoculated into YPD medium and cultured in a constant temperature shaker at 30°C and 150 r / min for 24 h. After two generations of continuous culture, the concentration was adjusted to 1×10 8 cfu / mL, and the Candida utilis seed solution was obtained; Lactobacillus plantarum was inoculated into MRS medium and cultured at 37°C for 24 h. After two generations of continuous culture, the concentration was adjusted to 1×10 8 cfu / mL, and Lactobacillus plantarum seed solution was obtained; S3. First stage fermentation The Candida utilis seed liquid was inoculated into the fermentation base material of the fungus bran at an inoculation rate of 6%, and the fermentation was carried out in a constant temperature incubator at 30°C for 60 hours with an oxygen flow rate of 8L / min and the material was turned once every 12 hours; S4. Second stage fermentation The Lactobacillus plantarum seed liquid was inoculated into the first stage fermentation product at an inoculum rate of 4%, mixed evenly and put into a respiratory fermentation bag, compacted to remove the air in the bag and sealed, and fermented at a constant temperature of 37° C. for 8 days to obtain the step-fermented bran feed.

2. The method for producing cattle and sheep feed by fermenting Pleurotus eryngii husk as raw material in steps according to claim 1, characterized in that: In S1, the preparation method of the fermented mushroom bran base material is: evenly mix the Pleurotus eryngii mushroom bran and molasses water in a solid-liquid ratio of 10g:1mL, and sterilize at 121°C for 40min; wherein the base material of the Pleurotus eryngii mushroom bran is: bagasse, corn cobs, corn flour, bran and light calcium.

3. The method for producing cattle and sheep feed by fermenting Pleurotus eryngii husk as raw material in steps according to claim 2, characterized in that: The brix of the molasses water is 16° Bx.

4. The method for producing cattle and sheep feed by fermenting Pleurotus eryngii husk as raw material in steps according to claim 1, characterized in that: In S2, the formula of YPD medium is: yeast powder 10.0 g, glucose 20.0 g, peptone 20.0 g, distilled water 1000 mL, pH 6.0 ± 0.2, sterilized at 121 °C for 15 min.

5. The method for producing cattle and sheep feed by fermentation stepwise using Pleurotus eryngii husk as raw material according to claim 1, characterized in that: In S2, the formula of MRS medium is: casein digest 10.0g, beef extract powder 10.0g, yeast extract powder 4.0g, triammonium citrate 2.0g, sodium acetate 5.0g, MgSO4·7H2O 0.2g, MnSO4·4H2O0.05g, dimethyl phosphate 2.0g, glucose 20.0g, Tween-80 1.0g, distilled water 1000mL, pH 5.7±0.2, sterilized at 121℃ for 15min.

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