Preparation method of hybrid pennisetum silage

Abstract

The invention belongs to the field of silage, and particularly relates to a preparation method of hybrid pennisetum silage, which comprises the following steps: S1, uniformly mixing a hybrid pennisetum raw material with lactobacillus plantarum LP276 and cellulase to obtain a silage raw material; s2, fermenting the silage raw materials at the temperature of 20-30 DEG C for not less than 45 days; wherein the lactobacillus plantarum LP276 is preserved in the Sichuan Province Microbial Resource Platform Culture Collection Center, and the preservation number is SICC 1.384. The preparation method provided by the invention breaks through the disadvantage that the traditional pennisetum alopecuroides silage is easy to decay, realizes the effects of effectively preserving the nutritional ingredients and improving the animal palatability and feeding value, can partially replace high-quality roughage such as silage corn and the like, and reduces the feeding cost of beef cattle and mutton sheep.

Description

Technical Field

[0001] The invention belongs to the field of silage, and particularly relates to a method for preparing hybrid Pennisetum silage. Background Art

[0002] Hybrid Pennisetum is one of the most widely planted forage grasses in southern my country. It grows fast, has high biomass, grows vigorously in summer, and its output accounts for more than 60% of the whole year. The seasonal imbalance is serious. Therefore, the processing and modulation of hybrid Pennisetum is particularly important. Silage is an ideal processing method that not only retains its nutrients, but also improves palatability and utilization rate. It is one of the effective means to solve the seasonal imbalance of forage production. However, the climate in the south is hot and humid, with abundant rainfall and relative air humidity of about 70% to 80%. Forage silage is easy to corrupt, which has become a bottleneck restricting the high-quality development of herbivorous animal husbandry in the region. At the same time, the moisture content of hybrid Pennisetum is as high as 80% to 85% when it is harvested, which is easy to cause Clostridium activity and produce a large amount of butyric acid, and silage is prone to failure.

[0003] In high temperature and high humidity areas, conventional lactic acid bacteria cannot improve the quality of silage, and silage is seriously corrupted. The development and application of excellent lactic acid bacteria can greatly reduce the corruption rate of hybrid Pennisetum silage, thereby improving its utilization rate and reducing environmental pollution.

[0004] Patent application with publication number CN117322529A (publication date: 2024-01-02) discloses the use of Lactobacillus rhamnosus in the preparation of hybrid Pennisetum silage, which has good inhibitory effects on Escherichia coli, Staphylococcus aureus, Listeria and aflatoxin, and improves the fermentation quality of hybrid Pennisetum silage. However, the microbial resources for hybrid Pennisetum silage in high temperature and high humidity areas are still in urgent need of development.

[0005] Lactobacillus plantarum LP276 is deposited in the Sichuan Microbial Resources Platform Culture Collection Management Center with a collection number of SICC 1.384. Currently, there is no report on the application of this strain in the preparation of hybrid Pennisetum silage. Summary of the invention

[0006] The purpose of the present invention is to provide a method for preparing hybrid Pennisetum silage in view of the above shortcomings, which overcomes the disadvantage that traditional Pennisetum silage is easily corrupted, achieves the effect of effectively preserving its nutritional components and improving its animal palatability and feeding value.

[0007] The objective of the present invention is achieved through the following technical solutions:

[0008] A method for preparing hybrid Pennisetum silage comprises the following steps:

[0009] S1. The hybrid Pennisetum raw material is mixed with Lactobacillus plantarum LP276 and cellulase to obtain silage raw material;

[0010] S2. fermenting the silage material at a temperature of 20 to 30° C. for not less than 45 days;

[0011] Among them, the Lactobacillus plantarum LP276 is preserved in the Sichuan Microbial Resources Platform Strain Collection Management Center with a preservation number of SICC 1.384.

[0012] In some preferred embodiments, the cellulase comprises an Acremonium cellulase.

[0013] In some preferred embodiments, in S1, the length of the hybrid Pennisetum raw material is 1 to 2 cm.

[0014] In some preferred embodiments, in S1, the content of Lactobacillus plantarum LP276 in the silage raw material is 5×10 5 CFU / g, and the content of the cellulase is 20U / g.

[0015] In some preferred embodiments, in S1, the moisture content of the silage material is 80%.

[0016] In some preferred embodiments, in S2, the fermentation is a sealed fermentation.

[0017] In some preferred embodiments, in S2, before fermentation, the silage material is pressed to a density of 750 kg / m 3 .

[0018] It should be noted that the present invention adds Lactobacillus plantarum LP276 to the hybrid Pennisetum raw material to increase the number of lactic acid bacteria, improve the relative abundance of Lactobacillus, promote lactic acid fermentation and reduce pH value, thereby improving the silage quality of the hybrid Pennisetum.

[0019] The cellulase treatment of Acremonium spp. enhanced the degradation of structural carbohydrates, provided more fermentation substrates for microorganisms, increased the relative abundance of Weissella, promoted the production of a large amount of lactic acid in the early stage of ensiling, and lowered the pH value, thereby improving the fermentation quality.

[0020] The beneficial effects of the present invention are:

[0021] 1. The preparation method of the present invention overcomes the shortcoming of the traditional Pennisetum silage being easy to corrupt, achieves the effect of effectively preserving its nutritional components and improving its palatability and feeding value for animals, and can partially replace high-quality roughage such as silage corn, thereby reducing the cost of raising beef cattle and sheep;

[0022] 2. The present invention has important theoretical significance and application value in overcoming the problem of unstable quality of high-moisture forage silage in hot and humid areas and promoting the development and utilization of bacterial enzyme compound additives in the area. DETAILED DESCRIPTION

[0023] The technical solution of the present invention is further described in detail below in conjunction with embodiments, but the protection scope of the present invention is not limited to the following.

[0024] Materials preparation:

[0025] For the hybrid Pennisetum raw material, mow the hybrid Pennisetum at 8-10 cm from the ground, and chop it with a scissors to 1-2 cm for later use.

[0026] Lactobacillus plantarum LP276, purchased from Sichuan Microbial Resources Platform Culture Collection Management Center, with the collection number SICC 1.384, provided in freeze-dried form, with a viable count greater than 1×10 11 CFU / g.

[0027] Lactobacillus rhamnosus was purchased from the Sichuan Microbial Resources Platform Strain Collection Management Center and provided in freeze-dried form.

[0028] Acremonium cellulase was purchased from Meiji Confectionery Co., Ltd., Tokyo, Japan, with a total enzyme activity greater than 1×10 3 U / g, mainly composed of α-galactosidase, β-glucanase, β-galactosidase, β-xylanase and α-isoprolinase.

[0029] Commercial cellulase preparation, purchased from Shanghai Yuanye Biotechnology Co., Ltd., with an enzyme activity of 5×10 4 U / g. .

[0030] Example 1

[0031] A method for preparing hybrid Pennisetum silage comprises the following steps:

[0032] S1. Activate the plant lactobacillus LP276 half a day before the start of the experiment, weigh a certain amount of the strain required for silage fermentation, add it to 2 ml of sterilized skim milk powder solution and activate it at room temperature for 2 hours, and weigh a certain amount of Acremonium cellulase and add it to distilled water and mix evenly. Mix the activated bacterial solution with the enzyme solution and add it to a spray pot, and evenly spray it on the hybrid Pennisetum raw material to obtain the silage raw material.

[0033] During the period, the spraying amount should be controlled so that the content of Lactobacillus plantarum LP276 in the silage raw material is 5×10 5 CFU / g, the cellulase content is 20U / g, and the moisture content is 80%.

[0034] S2. The silage material is loaded into a silage tank (1L) and compacted and sealed with tape, with a compaction density of 750kg / m 3 (about 750g per can), and silage is completed after fermentation at 30℃ for 45 days.

[0035] Comparative Example 1

[0036] A method for preparing hybrid Pennisetum silage comprises the following steps:

[0037] Spray distilled water on the hybrid Pennisetum raw material to make its moisture content 80%, then put it into a silage tank (1L), compact it and seal it with tape, and the compaction density is 750kg / m 3 (about 750g per can), and silage is completed after fermentation at 30℃ for 45 days.

[0038] The difference between Comparative Example 1 and Example 1 is that no microbial agent and cellulase are added.

[0039] Comparative Example 2

[0040] A method for preparing hybrid Pennisetum silage is different from Example 1 only in that Lactobacillus plantarum LP276 is replaced by Lactobacillus rhamnosus, and other aspects such as lactic acid bacteria content, cellulase type and content, fermentation conditions, etc. are the same.

[0041] Comparative Example 3

[0042] A method for preparing hybrid Pennisetum silage is different from Example 1 only in that the Acremonium cellulase is replaced with a commercial cellulase preparation on the market, and other aspects such as strain type, content and fermentation conditions are the same.

[0043] In the above examples, during the silage process, 20 g of silage was taken regularly using the five-point sampling method, 180 ml of distilled water was added, and after being shaken evenly by hand, it was placed in a 4°C refrigerator for extraction for at least 24 hours, and then filtered with 4 layers of gauze and qualitative filter paper. The pH value of the filtrate was determined using a glass electrode pH meter (pHS-3C, Shanghai, China). After the determination, the filtrate was treated with a 0.22 μm pore size water microporous filter membrane, and the organic acid (lactic acid, acetic acid) content was determined using a Shimadzu GC-14 high performance liquid chromatography (Japan), and the following Tables 1 to 3 were obtained.

[0044] Table 1 Effects of additives on pH value of hybrid Pennisetum silage

[0045]

[0046]

[0047] According to Table 1, compared with the comparative example, in Example 1, the pH value rapidly dropped to below 4.0 after 3 days of fermentation, which was conducive to the normal reproduction and fermentation of lactic acid bacteria in silage. Lactic acid bacteria fermented to produce lactic acid, which further reduced the pH value of the feed, thereby inhibiting the growth of harmful microorganisms such as mold and spoilage bacteria, and reducing the corruption and deterioration of the feed.

[0048] Table 2 Effect of additives on lactic acid content (g / kg DM) in hybrid Pennisetum silage

[0049]

[0050] Table 3 Effect of additives on acetic acid content (g / kg DM) in hybrid Pennisetum silage

[0051]

[0052] Organic acids (such as lactic acid, acetic acid, etc.) are important products in the fermentation process of silage. They not only reflect the fermentation degree of the feed, but also directly affect the quality and nutritional value of the feed. According to Tables 2 and 3, after the fermentation, the lactic acid and acetic acid contents of Example 1 are higher than those of the comparative example, which means that the silage fermentation of Example 1 is more complete and the quality is better.

[0053] During the ensiling process, samples were collected regularly and the ammonia nitrogen content was calculated based on the phenol-sodium hypochlorite colorimetric method to obtain the following Table 4.

[0054] Table 4 Effect of additives on ammonia nitrogen content (g / kg TN) in hybrid Pennisetum silage

[0055]

[0056] According to Table 4, the ammonia nitrogen content in the silage obtained in Example 1 is lower than that in the comparative example. It should be noted that the ammonia nitrogen content directly reflects the degree of degradation of protein and amino acids in the silage. The higher the ammonia nitrogen content, the more protein is decomposed, which usually means that the quality of the silage is poor. In addition, excessively decomposed protein may also lead to changes in feed structure and affect the digestibility of livestock.

[0057] The neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents of silage during fermentation were determined using the Ankom fully automatic fiber analyzer to obtain the following Tables 5 and 6.

[0058] Table 5 Effect of additives on neutral detergent fiber content (g / kg DM) in hybrid Pennisetum silage

[0059]

[0060] Table 6 Effect of additives on the acid detergent fiber content (g / kgDM) in hybrid Pennisetum silage

[0061]

[0062] According to Tables 5 and 6, after the fermentation, the acid detergent fiber content and the neutral detergent fiber content in the silage obtained in Example 1 were lower than those in the comparative example. Acid detergent fiber and neutral detergent fiber are mainly composed of cellulose, hemicellulose and lignin, which are difficult to be enzymatically hydrolyzed and utilized in livestock. The higher the content, the lower the digestibility, that is, the lower the silage quality.

[0063] During the ensilage process, the anthrone-sulfuric acid colorimetric method was used to determine the content of soluble carbohydrates (WSC) in the silage, and the following Table 7 was obtained.

[0064] Table 7 Effect of additives on soluble carbohydrate content (g / kgDM) in hybrid Pennisetum silage

[0065]

[0066]

[0067] According to Table 7, the soluble carbohydrate content in the silage obtained in Example 1 is higher than that in other comparative examples. Soluble carbohydrates refer to carbohydrates soluble in water, such as sucrose, fructose, glucose, etc., which are important energy sources for livestock. Therefore, a high content of soluble carbohydrates helps to improve the nutritional value of silage and meet the energy needs of livestock.

[0068] The above is only a preferred embodiment of the present invention. It should be understood that the present invention is not limited to the form disclosed herein, and should not be regarded as excluding other embodiments, but can be used in various other combinations, modifications and environments, and can be modified within the scope of the concept described herein through the above teachings or the technology or knowledge of the relevant field. The changes and modifications made by those skilled in the art shall not deviate from the spirit and scope of the present invention, and shall be within the scope of protection of the claims attached to the present invention.

Claims

1. A method for preparing hybrid Pennisetum silage, characterized in that: The following steps are involved: S1. The hybrid Pennisetum raw material is mixed with Lactobacillus plantarum LP276 and cellulase to obtain silage raw material; S2. fermenting the silage material at a temperature of 20 to 30° C. for not less than 45 days; Among them, the Lactobacillus plantarum LP276 is preserved in the Sichuan Microbial Resources Platform Strain Collection Management Center with a preservation number of SICC 1.

384.

2. The method for preparing hybrid Pennisetum silage according to claim 1, characterized in that: In S1, the cellulase comprises Acremonium cellulase.

3. The method for preparing hybrid Pennisetum silage according to claim 1, characterized in that: In S1, the length of the hybrid Pennisetum raw material is 1 to 2 cm.

4. The method for preparing hybrid Pennisetum silage according to claim 1, characterized in that: In S1, the content of Lactobacillus plantarum LP276 in the silage raw material is 5×10 5 CFU / g, and the content of the cellulase is 20U / g.

5. The method for preparing hybrid Pennisetum silage according to claim 1, characterized in that: In S1, the moisture content of the silage raw material is 80%.

6. The method for preparing hybrid Pennisetum silage according to claim 1, characterized in that: In S2, the fermentation is sealed fermentation.

7. The method for preparing hybrid Pennisetum silage according to claim 1, characterized in that: In S2, before fermentation, the silage material is pressed to a density of 750 kg / m 3 .

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