Functional forage grass with effects of improving mastitis and increasing milk yield as well as preparation method and application of functional forage grass

Abstract

The invention relates to functional forage grass with the effects of improving mastitis and increasing milk yield and a preparation method and application thereof, the functional forage grass comprises a forage grass matrix material and a nutrient material, and the functional forage grass is prepared by first-stage fungus aerobic fermentation and second-stage lactic acid bacteria and saccharomycetes anaerobic fermentation. Therefore, the functional forage grass is prepared by a first aerobic and second anaerobic two-stage fermentation method, can generate organic acid in a good proportion, has the characteristics of high lactic acid content, high protein content, high rumen degraded protein content, high dry matter content and the like, and can effectively inhibit the growth of other microorganisms; and the purpose of maintaining the stability of the unsealed functional forage grass is achieved. The functional forage grass is used for preparing feed products capable of improving mastitis of herbivorous animals and increasing milk yield of the herbivorous animals.

Description

Technical Field

[0001] The present invention belongs to the technical field of feed, and particularly relates to a functional forage grass with the functions of improving the occurrence of mastitis and increasing milk production, and a preparation method and use thereof.

[0002] Prior Art

[0003] In animal husbandry, ruminant feed is the main food component of herbivores, accounting for more than half of the daily diet of herbivores. Therefore, the quality and supply stability of ruminant feed directly affect the health and production of herbivores. To overcome the influence of seasons on ruminant feed, a silage feed has been developed, which is made from green crops or agricultural by-products after being sealed and fermented.

[0004] During the fermentation process of silage feed, carbon dioxide produced by the fermentation of lactic acid bacteria decomposing sugars is used to expel air to form an anaerobic fermentation environment, and the lactic acid secreted by lactic acid bacteria makes the feed weakly acidic (pH value 3.5 - 4.2), which can effectively inhibit the growth of other microorganisms; finally, the lactic acid bacteria are inhibited by the lactic acid produced by themselves and terminate the fermentation, enabling the feed to enter a stable storage state.

[0005] Therefore, silage feed has the following advantages: (1) Compared with hay, the harvesting is not severely affected by weather conditions, which helps to reduce losses from harvest to storage; (2) The production process of silage feed can be mechanized, saving labor and time; (3) Silage feed can not only maintain good quality, but also effectively manage forage production and increase the harvest in the hot season; (4) Due to the appropriate moisture content, the acceptance and feed intake of animals for silage feed are improved; (5) The feeding amount and quality are stable, and the production performance of animals is stable. Therefore, from the perspectives of nutrition and management, if high-quality ruminant feed can be timely modulated into silage, the nutrient loss is less, the yield and quality can be stabilized, and it has practical significance in production and health management.

[0006] However, there are also the following disadvantages when modulating silage feed: (1) Sufficient site, raw materials and capital supply are required; (2) The quality of raw materials is not easy to control; (3) Great losses occur when the container is not properly selected; (4) Quality losses caused by improper management during the modulation process or after opening; (5) Animal production capacity and health problems caused by improper diet formulation. Among them, it is particularly noteworthy that once the silage bag is opened, the internal microbial environment will be affected by oxygen and microorganisms in the air, which is prone to cause fermentation failure, mildew or other microbial contaminations, resulting in nutrient loss and quality decline of the silage feed. In addition, although the existing silage feed can affect the rumen fermentation process, its function has not been reported to be able to simultaneously significantly increase the milk production of herbivores and effectively reduce mastitis inflammation, indicating that the comprehensive regulation of the physiological functions of herbivores needs to be further improved and perfected.

[0007] In summary, silage has an indispensable position in animal husbandry, but it also faces a series of challenges and problems, especially in terms of preservation after opening, which needs to be further solved to give full play to the advantages of silage while reducing its disadvantages. Summary of the Invention

[0008] In view of the above technical problems, the present invention provides a functional forage grass with the functions of improving the occurrence of mastitis and increasing milk production, and its preparation method and uses. The functional forage grass includes a forage grass matrix material and a nutritional material, or the functional forage grass is composed of a forage grass matrix material and a nutritional material, and is prepared by first performing a first-stage aerobic fermentation with fungi and then performing a second-stage anaerobic fermentation with lactic acid bacteria and yeasts. Thus, by using a two-stage fermentation method of first aerobic and then anaerobic to prepare the functional forage grass, the functional forage grass decomposes the forage grass matrix material in the first-stage aerobic fermentation and produces functional components such as mushroom polysaccharides, ergothioneine (EGT), ergosterol, and polyphenols, and decomposes the remaining sugars of the first fermentation product formed in the first-stage aerobic fermentation in the second-stage anaerobic fermentation and produces organic acids in a good proportion, so that the functional forage grass of the present invention has high contents of lactic acid, total protein, and rumen-degradable protein at the same time, and has excellent characteristics such as a higher dry matter content than ordinary silage. The functional forage grass of the present invention decomposes the remaining monosaccharides and disaccharides that are easily utilized by the growth of other microorganisms in the second-stage anaerobic fermentation, so that after the functional forage grass of the present invention is opened, other microorganisms are not easy to grow, achieving the purpose of maintaining the stability of the functional forage grass after opening.

[0009] Based on the above purposes, the present invention provides a functional forage grass, which includes a raw material containing a forage grass matrix material and a nutritional material, and the raw material is prepared into the functional forage grass by first performing a first-stage aerobic fermentation with fungi and then performing a second-stage anaerobic fermentation with lactic acid bacteria and yeasts. Alternatively, the present invention provides a functional forage grass, which includes a raw material composed of a forage grass matrix material and a nutritional material, and the raw material is prepared into the functional forage grass by first performing a first-stage aerobic fermentation with fungi and then performing a second-stage anaerobic fermentation with lactic acid bacteria and yeasts.

[0010] According to an embodiment of the present invention, the total organic acid content of the functional forage grass reaches 160 grams (g) or more per kilogram (i.e., the functional forage grass per kilogram contains equal to or more than 160 g of organic acids), and the organic acids include lactic acid, acetic acid, and butyric acid.

[0011] In an embodiment of the present invention, the functional forage may contain 160 g, 170 g, 180 g, 190 g, 200 g, 210 g, 220 g, 230 g, 240 g, 250 g, 260 g, 270 g, 280 g, 290 g, 300 g, 310 g, or 320 g of total organic acid content (including lactic acid, acetic acid, and butyric acid) per kilogram; the total organic acid content contained in the functional forage per kilogram may be within the range formed by any two of the above values, but is not limited thereto.

[0012] According to the implementation scheme of the present invention, gramineous plants can be selected as the forage matrix material. Specifically, the forage matrix material can be selected from one or any combination of Pennisetum alopecuroides, Pennisetum purpureum, Sorghum sudanense, Avena sativa, Agropyron cristatum, and Trifolium alexandrinum. In an embodiment of the present invention, the forage matrix material can be Agropyron cristatum.

[0013] According to the implementation scheme of the present invention, carbon element (C) materials, nitrogen element (N), vitamins, minerals, etc. can be selected as nutrient materials. In an embodiment of the present invention, the nutrient materials can be selected from one or any combination of soybean powder, wheat bran, and corncob as the nutrient materials. In another embodiment of the present invention, the nutrient materials can be a combination of corncob, soybean powder, and wheat bran.

[0014] According to the implementation scheme of the present invention, calculated based on the total weight of the raw materials, the total water content of the raw materials can be 45 wt% to 65 wt%, and the total dry matter content of the raw materials can be 35 wt% to 55 wt%; calculated based on the total dry matter content of the raw materials being 100 wt%, 40 wt% to 60 wt% of the total dry matter content of the raw materials can come from forage, 10 wt% to 20 wt% can come from soybean powder, and 30 wt% to 40 wt% can come from wheat bran; among them, in the raw materials selected in the embodiment of the present invention, the water content of the forage is about 10 wt% to 20 wt%, the water content of the soybean powder is about 5 wt% to 15 wt%, and the water content of the wheat bran is about 5 wt% to 15 wt%.

[0015] According to the implementation scheme of the present invention, the functional forage may contain 160 g to 320 g of total organic acid content per kilogram; the organic acids in the functional forage may include lactic acid, acetic acid, and butyric acid.

[0016] In an embodiment of the present invention, the functional forage may contain up to 210 grams of lactic acid (g lactic acid / kg functional forage) content per kilogram. In an embodiment of the present invention, the functional forage may contain 110 g, 120 g, 130 g, 140 g, 150 g, 160 g, 170 g, 180 g, 190 g, 200 g, or 210 g of lactic acid content per kilogram; the lactic acid content contained in the functional forage per kilogram may be within the range formed by any two of the above values, but is not limited thereto.

[0017] In the embodiments of the present invention, the functional forage may contain up to 74 grams of acetic acid per kilogram (g acetic acid / kg functional forage). In one embodiment of the present invention, the functional forage may contain 8g, 10g, 15g, 20g, 25g, 30g, 35g, 40g, 45g, 50g, 55g, 60g, 65g, 70g or 74g of acetic acid; the acetic acid content contained in the functional forage per kilogram may be within the range formed by any two of the above values, but is not limited thereto.

[0018] In the embodiments of the present invention, the functional forage may contain up to 40 grams of butyric acid per kilogram (g butyric acid / kg functional forage). In one embodiment of the present invention, the functional forage may contain 14g, 15g, 20g, 25g, 30g, 35g or 40g of acetic acid; the acetic acid content contained in the functional forage per kilogram may be within the range formed by any two of the above values, but is not limited thereto.

[0019] According to the embodiments of the present invention, the dry matter content of the functional forage may be up to 42 wt%. In one embodiment of the present invention, the dry matter content of the functional forage may be 26 wt%, 27 wt%, 28 wt%, 29 wt%, 30 wt%, 31 wt%, 32 wt%, 33 wt%, 34 wt%, 35 wt%, 36 wt%, 37 wt%, 38 wt%, 39 wt%, 40 wt%, 41 wt% or 42 wt%; the dry matter content of the functional forage may be within the range formed by any two of the above values, but is not limited thereto.

[0020] According to the embodiments of the present invention, the rumen crude protein digestibility of the functional forage can reach 16%. In one embodiment of the present invention, the rumen crude protein digestibility of the functional forage may be 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15% or 16%; the rumen crude protein digestibility of the functional forage may be within the range formed by any two of the above values, but is not limited thereto.

[0021] According to the embodiments of the present invention, calculated based on the total protein content of the functional forage, the rumen degradable protein content of the functional forage can reach 77% of the total protein. In one embodiment of the present invention, the rumen degradable protein content of the functional forage may be 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79% or 80%; the rumen degradable protein content of the functional forage may be within the range formed by any two of the above values, but is not limited thereto.

[0022] Another object of the present invention is to provide a method for preparing the above-mentioned functional forage grass, which includes a first-stage aerobic fermentation and a second-stage anaerobic fermentation. Among them,

[0023] The steps of the first-stage aerobic fermentation include:

[0024] Cut the forage grass into forage matrix materials;

[0025] Put in nutrient materials and evenly mix them with the forage matrix materials and adjust the moisture content to form a mixed material;

[0026] After sterilizing the mixed material, add fungi to the mixed material;

[0027] After the mixed material has been colonized by the fungi, place it in an aerobic and constant room temperature environment for aerobic fermentation for 1 to 2 months to obtain a first fermentation product;

[0028] The steps of the second-stage anaerobic fermentation include:

[0029] Add yeast and lactic acid bacteria to the first fermentation product, then mix and stir and adjust the moisture content to form forage materials;

[0030] Pack the forage materials to form an anaerobic fermentation environment, and carry out anaerobic fermentation for 7 to 28 days to obtain the functional forage grass.

[0031] According to the implementation scheme of the present invention, in the first-stage aerobic fermentation, based on the weight of the mixed material, the addition amount of the fungi is 0.1wt% to 10wt%; the fungi are one or a combination of edible fungi and medical fungi. In an embodiment of the present invention, the fungi are edible fungi, such as mushroom fungi. In an embodiment of the present invention, the fungi are Pleurotus eryngii, purchased from the Food Industry Research Institute (BCRC36907), but the fungi described in the present invention are not limited to this strain. In the implementation scheme of the present invention, the term "colonized by the fungi" refers to mycelial growth.

[0032] According to the embodiments of the present invention, in the second-stage anaerobic fermentation, based on the weight of the first fermentation product, the addition amount of the yeast is 0.1 wt% to 10 wt%, and the addition amount of the lactic acid bacteria is 0.1 wt% to 10 wt%. In an embodiment of the present invention, the lactic acid bacteria are Lactobacillus plantarum (L.P.) and / or Lactobacillus buchneri (L.B.), purchased from the Food Industry Research Institute (BCRC 10069D and BCRC 17760), but the lactic acid bacteria described in the present invention are not limited to these strains; the yeast is Saccharomyces cerevisiae (S.C.), purchased from the Food Industry Research Institute (BCRC 20262); however, the yeast described in the present invention is not limited to this strain.

[0033] According to the embodiments of the present invention, in the first-stage aerobic fermentation, the mixed material can be sterilized by using a sterilization kettle, steam, microwave or other means to achieve the purpose of sterilization.

[0034] According to the embodiments of the present invention, in the second-stage anaerobic fermentation, the forage material is made into grass balls by using a baling and wrapping machine to wrap a multi-layer plastic film of low-density polyethylene (LLDPE) (thickness 0.025 mm) to form an anaerobic fermentation environment, and the functional forage is obtained after anaerobic fermentation for 7 to 28 days.

[0035] According to the embodiments of the present invention, in the first-stage aerobic fermentation, after the mixed material is colonized by bacteria, it can be fermented in a constant room temperature environment of 10°C to 30°C. Preferably, after the mixed material is colonized by bacteria, it can be fermented in a constant room temperature environment of 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C or 30°C; but not limited thereto. Each of the above specific values can be used as the end point value of another range.

[0036] Another object of the present invention is to provide the use of the functional forage as described above or the functional forage prepared by the preparation method as described above for preparing a feed product for improving the mastitis of herbivores. Thus, the functional forage of the present invention combines the principles of nutrition and natural therapy, so that when herbivores eat the functional forage of the present invention, they can naturally obtain the effect of non-drug anti-inflammatory and avoid the dependence and possible side effects brought by traditional drug treatment.

[0037] Another object of the present invention is to provide the use of the functional forage as described above or the functional forage prepared by the preparation method as described above for preparing a feed product for increasing the milk production of herbivores.

[0038] According to the embodiments of the present invention, calculated by the total weight percentage (100 wt%) of the feed product, the addition amount of the functional forage in the feed product can be 8 wt% to 17.5 wt%. In an embodiment of the present invention, the addition amount of the functional forage in the feed product can be 8.0 wt%, 8.47 wt%, 8.5 wt%, 9.0 wt%, 9.5 wt%, 10.0 wt%, 10.5 wt%, 11.0 wt%, 11.5 wt%, 12.0 wt%, 12.5 wt%, 13.0 wt%, 13.5 wt%, 14.0 wt%, 14.5 wt%, 15.0 wt%, 15.5 wt%, 16.0 wt%, 16.5 wt%, 16.98 wt%, 17.0 wt% or 17.5 wt%; the addition amount of the functional forage in the feed product can be within the range formed by any two of the above values, but is not limited thereto.

[0039] In an embodiment of the present invention, the feed product can be a commercial Total Mixed Ration (TMR) formula.

[0040] Brief Description of the Drawings

[0041] Figure 1 It is a graph showing the change of rumen crude protein digestibility over time of the products of Example 2 (E2) and Example 3 (E3) of the present invention.

[0042] Figure 2 It is a graph of the milk yield change data of three groups of cows (control group, low substitution group, high substitution group) in Test Example 5 of the present invention within four weeks. Detailed Description of the Embodiments

[0043] In order to understand the technical features, content, advantages and achievable effects of the present invention, the present invention will be described in detail below in conjunction with the form of the attached drawings. The drawings used herein are only for illustration and auxiliary description of the specification, and may not be the true ratio and precise configuration after the implementation of the present invention. Therefore, the present invention should not be interpreted and limited to the scope of rights in actual implementation based on the ratio and configuration relationship of the attached drawings, unless otherwise stated in advance.

[0044] Preparation Example 1: Functional Forage

[0045] The forage matrix material in the raw materials of this preparation example is wheatgrass, and the nutritional materials are soybean powder, wheat bran, and corncob. Calculated based on the total weight of the raw materials, the total water content of the raw materials is about 55 wt%, and the total dry matter content of the raw materials is about 45 wt%; calculated based on the total dry matter content of the raw materials being 100 wt%, about 41 wt% of the total dry matter content of the raw materials comes from forage, about 14 wt% comes from soybean powder, about 20 wt% comes from wheat bran, and about 25 wt% comes from corncob; among them, in the raw materials selected in this preparation example, the water content of the forage is about 10 wt%, the water content of the soybean powder is about 8 wt%, the water content of the wheat bran is about 10 wt%, and the water content of the corncob is about 10 wt%.

[0046] The preparation method of this preparation example includes the first aerobic fermentation using fungi and the second anaerobic fermentation using lactic acid bacteria and yeast, where:

[0047] The steps of the first aerobic fermentation include:

[0048] Chop the forage into pieces of 2 to 4 cm in size as the forage matrix material;

[0049] Put the nutritional materials into the forage matrix material, mix evenly, and adjust the water content to 55% wt% to form a mixed material;

[0050] After sterilizing the mixed material using a sterilization kettle, add fungi to the mixed material; the fungi are Pleurotus eryngii; calculated based on the weight of the mixed material, the addition amount of the fungi is about 2 wt%;

[0051] After the mixed material is colonized by the fungi, place it in an aerobic and constant temperature environment of 22°C to 25°C for aerobic fermentation for 1 to 2 months to obtain the first fermentation product;

[0052] The steps of the second anaerobic fermentation include:

[0053] Add yeast and lactic acid bacteria to the first fermentation product, mix and stir, and adjust the water content to 60 wt% to form a forage material; the lactic acid bacteria are Lactobacillus plantarum (L.P.) and Lactobacillus buchneri (L.B.), and the yeast is Saccharomyces cerevisiae (S.C.); calculated based on the weight of the first fermentation product, the addition amount of the lactic acid bacteria is about 0.3 wt%, and the addition amount of the yeast is about 0.3 wt%;

[0054] Using a baling and wrapping machine, the forage material is wrapped with a multi-layer plastic film of low-density polyethylene (LLDPE) (thickness 0.025 mm) to form a grass ball. After anaerobic fermentation for 7 to 28 days, the functional forage is obtained.

[0055] Preparation Example 2: Corn Silage

[0056] The corn silage in this preparation example was purchased from Jufeng Enterprise. The preparation method steps of this corn silage include:

[0057] (1) Moisture content of raw materials and pretreatment: The whole plant corn is harvested from the milk-ripe stage to the dough-ripe stage; after harvesting the ear, 1 / 2 of the green leaves are retained on the straw. After the fresh corn straw is sun-dried for 1 - 2 days, and when the moisture content of the silage raw materials is ensured to be 60% - 75%, straw silage is carried out. The whole plant corn straw for silage is cut short to about 3 cm.

[0058] (2) Making the mixed material: Add silage additives (microbial inoculants) to improve the silage process and enhance the silage quality.

[0059] (3) Packing in an anaerobic environment: During the process of preparing the silage material, it is necessary to compact it as soon as possible and shorten the exposure time of the raw materials in the air. Then, using a baling and wrapping machine, the forage material is wrapped with a multi-layer plastic film of low-density polyethylene (LLDPE) (thickness 0.025 mm) to form a grass ball.

[0060] (4) Placing for fermentation and ripening: When the grass ball made produces lactic acid and ferments to maturity after 40 - 60 days of silage, the corn silage is obtained.

[0061] Examples 1 to 4: Functional Forage

[0062] Select the product obtained in the aforementioned Preparation Example 1, and according to the fermentation days (7 days, 14 days, 21 days, 28 days) of the second anaerobic fermentation, the functional forages of Examples 1 to 4 (E1 to E4) of the present invention are formed.

[0063] In the examples of the present invention, the product obtained by the first aerobic fermentation is the first fermentation product, and the product obtained by the second anaerobic fermentation of the first fermentation product wrapped with a multi-layer plastic film of low-density polyethylene (LLDPE) (thickness 0.025 mm) using a baling and wrapping machine to form a grass ball is the second fermentation product; the fermentation days of the aforementioned second anaerobic fermentation refer to the total number of days from the completion of packing and sealing of the first fermentation product to the opening and taking out of the second fermentation product.

[0064] Comparative Example 1: Corn Silage

[0065] Select the product obtained in the aforementioned Preparation Example 2 as the corn silage of Comparative Example 1.

[0066] Test Example 1: Determination of Fermentation Quality of Functional Forage

[0067] The analysis methods for the dry matter (DM) content, crude protein (CP) content, pH value, ammonia nitrogen (NH3-N) content, lactic acid content, acetic acid content, and butyric acid content of the functional forages in the aforementioned Examples 1 to 4 and the corn silage in Comparative Example 1 are as follows, and the results are shown in Table 1.

[0068] 〔Dry matter (DM) content, crude protein (CP) content〕

[0069] Taking the functional forages in Examples 1 to 4 and the corn silage in Comparative Example 1 as the test samples, each test sample was placed in a fan-assisted oven and dried at 65 °C for 48 hours. By comparing the weights of each test sample before and after drying, the dry matter content ratio of each test sample was obtained, and then the dry matter recovery rate of the test sample for 90 days was estimated by multiplying the weights of each test sample before and after drying by its dry matter content ratio. Among them, the determination methods for the dry matter (DM) content and crude protein (CP) content are as described by Huisden [1] et al.

[0070] 〔Dry matter rumen digestibility (DMD), neutral detergent fiber rumen digestibility (NDFD)〕

[0071] Taking the functional forages in Examples 1 to 4 and the corn silage in Comparative Example 1 as the test samples, the neutral detergent fiber (NDF) and acid detergent fiber (ADF) of each test sample were determined as described by Van Soest et al. [2]. According to the dry matter content and neutral detergent fiber content in the test samples, the in vitro true DM disappearance (IVDMD) and in vitro neutral detergent fiber digestibility (IVNDFD) were determined. It was carried out by in vitro digestion for 48 hours, and the water soluble carbohydrate (WSC) was determined by the anthrone method [3] after extraction with 80% ethanol. The determination results of the in vitro dry matter rumen digestibility (IVDMD) and neutral detergent fiber rumen digestibility (IVNDFD) are shown in Table 1 below.

[0072] 〔pH value〕

[0073] The functional forages of Examples 1 to 4 and the corn silage of Comparative Example 1 were used as test samples. 20 g of each test sample was taken and 200 mL of cold (4 °C) deionized water was added thereto, and homogenized in a blender for 60 seconds (30 seconds for each homogenization, homogenized twice, with an interval of 30 seconds in between) to form a homogeneous suspension. Then, the homogeneous suspension was centrifuged at a centrifugal force of 12,500×g for 20 minutes, the supernatant was collected, and the pH value was measured with a pH meter. The measurement results of the pH value are shown in Table 1 below.

[0074] 〔Lactic acid, volatile fatty acid (VFA) and ammonia nitrogen (NH3-N)〕

[0075] The functional forages of Examples 1 to 4 and the corn silage of Comparative Example 1 were used as test samples. The supernatant used for measuring the pH value was filtered through a 0.22 μm disc filter, and the filtrate was collected for the determination and analysis of lactic acid, volatile fatty acid (VFA) and ammonia nitrogen (NH3-N). Among them, high performance liquid chromatography (the chromatographic column was Rezex ROA - organic acid H + (8%), 300 mm×7.8 mm, Phenomenex, USA) was used to measure the concentrations of lactic acid and volatile fatty acid [4]; and, the ammonia nitrogen content was measured by colorimetry according to Weatherburn [5].

[0076] As can be seen from the measurement results in Table 1, compared with the corn silage of Comparative Example 1 (C1), the dry matter (DM) contents of the functional forages of Examples 1 to 4 (E1 to E4) were all higher than 1.5 times the dry matter content of Comparative Example 1; the crude protein (CP) contents of Examples 1 to 4 were all higher than that of Comparative Example 1, and the protein decreased with the increase in the number of days of the second-stage anaerobic fermentation.

[0077] In the analysis of the ammonia nitrogen (NH3-N) content of the functional forage grass of the present invention, for the forage grass sample to be tested, after first adding distilled water and mixing (sample: water = 1:10), it is stirred with a juice extractor, and then the filtrate is taken to measure the ammonia nitrogen concentration and moisture content. Finally, the ammonia nitrogen (NH3-N) concentration per gram of the original forage grass sample to be tested is deduced (unit: mg / g fresh sample). The analysis results show that the ammonia nitrogen (NH3-N) content of the functional forage grass in Examples 1 to 4 is all lower than that of the corn silage in Comparative Example 1. In the part of the organic acid (lactic acid, acetic acid, butyric acid) content, it is calculated based on how many grams (g) of organic acid are contained in each kilogram (kg) of the functional forage grass of the examples and the corn silage of the comparative example (unit: g / kg). The functional forage grass in Examples 1 to 4 is the product of the second-stage anaerobic fermentation for 7 days, 14 days, 21 days, and 28 days in sequence, and the corn silage in Comparative Example 1 is the product prepared by anaerobic fermentation for 50 days throughout the process. Among them, when fermented to 14 days (E2), the functional forage grass prepared by the present invention has reached the highest lactic acid and acetic acid yields, and for Examples 1 to 4 (E1 to E4), whether it is the total organic acid content, lactic acid content, acetic acid content, or butyric acid content, they are all higher than the corresponding acid contents in Comparative Example 1.

[0078] Table 1: Determination results of the dry matter content, crude protein content, pH value, ammonia nitrogen content (NH3-N), lactic acid, acetic acid, and butyric acid contents of the functional forage grass in Examples 1 (E1) to 4 (E4) of the present invention and the corn silage in Comparative Example 1. (N.D. means the corresponding substance content was not measured)

[0079]

[0080] As can be seen from the determination results of Test Example 1 above, the functional forage grass (E1 to E4) of the present invention has the following effects compared with ordinary corn silage (C1):

[0081] (1) The anaerobic fermentation time of the functional forage grass (E1 to E4) of the present invention is short. It only takes about 14 days in the second-stage anaerobic fermentation to reach the highest acid production, which helps to improve the product stability after the functional forage grass is opened.

[0082] (2) The ammonia nitrogen concentration of the functional forage grass (E1 to E4) of the present invention is low, which can avoid the production of ammonia smell in the functional forage grass product and also avoid affecting the pH value of the functional forage grass of the present invention, so that the pH value can be maintained under a lower acidic condition, maintaining the antibacterial effect and product stability.

[0083] (3) The functional forage grass (E1 to E4) of the present invention contains more lactic acid and acetic acid than ordinary corn silage, and has the effect of inhibiting the growth of miscellaneous bacteria during the fermentation process; in addition, due to the rapid acid production of the functional forage grass of the present invention, it can maintain the functional forage grass in an acidic environment with a low pH value, effectively inhibiting bacteria and being durable and easy to store.

[0084] (4) Compare the quality of the functional forage grasses (E1 to E4) of the present invention with that of ordinary corn silage (C1) under anaerobic conditions. The ammonia-nitrogen concentration measured for the functional forage grasses of the present invention is low, the protein degradation is less, the fermentation loss is low, and the dry matter ratio is high. Therefore, compared with the corn silage (C1) made from ordinary green-cut corn, the dry matter ratio of the functional forage grasses of the present invention can be 1.5 times higher than that of the green-cut corn silage, and it has a higher nutritional value.

[0085] Test Example 2: Determination of the rumen degradation efficiency of functional forage grasses

[0086] The analysis method for the rumen degradation efficiency of the functional forage grasses in the foregoing embodiments and the corn silage in the comparative examples is as follows, and the results are shown in Figure 1 , Table 2 and Table 3.

[0087] Use the functional forage grasses of Examples 2 and 3 (E2 and E3) and the corn silage (C1) of Comparative Example 1 as the test samples. Referring to the method of Nocek (1985) [8] in this test example, the in situ rumen test is used to evaluate the crude protein (CP) digestibility and the proportion of rumen degradable protein (RDP) of the functional forage grasses prepared by the two-stage fermentation of the present invention.

[0088] A brief description of the process for determining the rumen crude protein digestibility is as follows: After drying each test sample, according to the experimental design, at each sampling time point, each test sample is divided into four digestion bags (purchased from ANKOM Technology; 5×10 cm; 50 μm pore size), and 2 grams of the dried test sample is placed in each bag, so that each test sample forms four replicate samples. After being marked, the digestion bags are tied tightly. First, soak them in warm water at 39 °C for 20 minutes, and then put them into the rumen of a windowed cow. At 0, 2, 4, 8, 12, 24, and 48 hours after being put in, four replicate samples of each test sample are taken out at each time point; each digestion bag taken out is rinsed on the surface with cold water to wash away the rumen fluid, and then put into an oven at 65 °C and dried for 48 hours. After drying, record the weight of the test sample to calculate its rumen crude protein digestibility, and the measured results are shown in the Figure 1 curve graph, and then estimate the results in Table 2 from the Figure 1 data. Combine the following formulas (1) and (2) to calculate the proportions of rumen degradable protein (RDP) and rumen undegradable protein (RUP) of Examples 2 and 3 (E2, E3) and Comparative Example 1 (C1) as shown in Table 3.

[0089] In this test example, the following formulas (1) and (2) are the calculation formulas for rumen degradable protein (RDP) and rumen undegradable protein (RUP), respectively:

[0090] Formula (1): RDP% = a + b * [c / (c + k)];

[0091] Formula (2): RUP% = 100% - RDP%;

[0092] In this formula (1), the outflow rate (k) is set to 6% (i.e., 0.06 / h), where the outflow rate refers to the percentage of rumen content flowing into the small intestine per hour.

[0093] From Figure 1 the graph showing the change of rumen crude protein digestibility (%) over time as shown, it can be seen that the rumen crude protein digestibility of the functional forage in Example 2 (E2) is slightly faster than that in Example 3 (E3), and the results in Table 2 can be obtained, and the values of a, b, c, etc. in this formula (1) can be obtained. Among them, the degradation mode of Table 2 and Figure 1 the data is a one-phase association model; the best fit values calculated by using non-linear regression for the data in Table 2 are as follows:

[0094] Y0 (= a), representing the protein that can be quickly decomposed;

[0095] Plateau, representing the total amount of decomposable protein;

[0096] K (= c), representing the average decomposition rate of protein during digestion, and the larger the value, the faster the digestion rate;

[0097] Half-life (half-time = t 1 / 2 ), representing the time required for the total amount to be decomposed to half;

[0098] Span (= b), representing the protein that is relatively difficult to decompose.

[0099] Table 2: Rumen crude protein digestibility (%) related parameters of the functional forage in Example 2 (E2) and Example 3 (E3) of the present invention in Test Example 2, obtained according to Figure 1 the curve.

[0100]

[0101] The RDP and RUP evaluation results of Examples 2 and 3 are shown in Table 3; among them, the RDP and RUP of Example 2 accounted for 77% and 23% of its total protein, respectively; the RDP and RUP of Example 3 accounted for 73% and 27% of its total protein, respectively. Compared with the RDP of 67% and RUP of 33% for general corn silage [9], it shows that the functional forage made by the two-stage fermentation of the present invention can provide a higher content of degradable protein than general corn silage.

[0102] Table 3: The measurement results of the respective contents of rumen degradable protein (RDP) and rumen undegradable protein (RUP) of the functional forage of Example 2 (E2) and Example 3 (E3) of the present invention and the corn silage of Comparative Example 1 (C1).

[0103] Sample ID to be measured E2 E3 C1 Fermentation days 14 days 21 days 50 days Rumen degradable protein (RDP) 77％ 73％ 67％ Rumen undegradable protein (RUP) 23％ 27％ 33％

[0104] Test Example 3: Aerobic stability test of functional forage

[0105] The analysis method for the aerobic stability of the functional forage of the foregoing examples and the corn silage of the comparative example is as follows, and the results are shown in Table 4. In this test example, the aerobic stability was measured according to the system described by Ashbell et al. [6].

[0106] Take 1000 grams of the test samples in the sampling bags respectively. After turning them loose, take 300 grams of them and put them into a plastic bottle with holes (the wet weight of each plastic bottle is 300 grams) to allow air to enter the bottle through the openings. Then, place a thermometer in each test sample to measure the central temperature of each test sample in the bottle, and record the temperature readings within 72 hours at intervals of every 12 hours. During the test period, a continuous recorder (HOBOMAX100, Smartec Scientific Corp., Taiwan Province, China) was used to record the ambient temperature every 15 minutes. In this test example, the definition of aerobic stability refers to the number of hours before the temperature of the test sample rises above 2°C higher than the ambient temperature [7]. The storage temperature during the test period of this test example was between 28°C and 34°C, and the relative humidity was above 80%.

[0107] In this test example, the functional forages of Examples 2 and 3 (E2 and E3) and the corn silage of Comparative Example 1 (C1) were used as test samples; among them, the functional forage of Example 2 was a sample with an anaerobic fermentation period of 14 days in the second stage, the functional forage of Example 3 was a sample with an anaerobic fermentation period of 21 days in the second stage, and Comparative Example 1 was a general corn silage with a full anaerobic fermentation period of 50 days.

[0108] Table 4: The temperature rise degrees at different cumulative time points measured within 72 consecutive hours of the functional forages of Example 2 (E2) and Example 3 (E3) of the present invention and the corn silage of Comparative Example 1 (C1).

[0109]

[0110] The evaluation results of the aerobic stability determination in this test example are shown in Table 4 above. In Examples 2 and 3 (E2, E3) of the present invention, the phenomenon of temperature rise of functional forage was detected only after 48 hours and 60 hours of testing respectively, and until the end of the 72-hour testing time, the temperature rise amplitudes of Examples 2 and 3 did not exceed 1 °C; while in Comparative Example 1 (C1), the temperature started to rise at 24 hours of testing and the amplitude was close to 1 °C, the temperature rise amplitude at 36 hours of testing was close to 2 °C, and the temperature rise amplitude at 48 hours of testing far exceeded 2 °C of the ambient temperature; as can be seen from the above, the functional forage prepared by the two-stage fermentation of the present invention has better aerobic stability than ordinary corn silage after being exposed to air.

[0111] Test Example 4: Determination of the effect of functional forage on improving mastitis

[0112] Since only a small number of somatic cell counts (SCC) are contained in normal milk; while the somatic cell count in the milk produced by dairy cows in the case of bacterial infection (mastitis) in the udder will increase significantly, therefore, the somatic cell count is usually used as a measure of milk quality, that is, a high somatic cell count represents low milk quality. Thus, the following determination method is used in this test example to test the improvement effect of the functional forage of the present invention on bovine mastitis.

[0113] 〔Experimental animals〕

[0114] In this test example, 21 Dutch lactating cows were divided into a control group, a low substitution group, and a high substitution group with 7 cows in each group for the test. The milk production of these cows was 29.28 ± 5.75 kg / day; the number of days in milk (DIM) was 163.3 ± 44.8 days; the average parity was 2.0 ± 1.1 parities.

[0115] 〔Experimental diet〕

[0116] In this test example, the functional forage of the present invention was used as the test sample, and the feed part in the formula of the commercial total mixed ration (TMR) fed to cows daily was partially replaced in different proportions, divided into zero substitution amount (control group), low substitution group, and high substitution group. Then, the milk of the test cows was collected and sent to the Hsinchu Branch of the Taiwan Livestock Research Institute in China to conduct DHI milk quality inspection with a milk composition and somatic cell analyzer (MilkoScanTM FT +, Denmark) to evaluate the somatic cell count and other milk quality indicators in the milk.

[0117] This experiment referred to the NRC 2001 to design a formula that meets the nutritional requirements of lactating cows, and feeding was carried out at 8:00 am and 1:00 pm every day. The commercial TMR formula and the replacement amount of functional forage used in this experimental example are shown in Table 5 below; this TMR formula mainly consists of a feed part (which may contain oat hay, corn silage, TMR No. 2), additives (which may contain baking soda, probiotics, mycotoxin adsorbents), and moisture. Calculated with the TMR formula as 100 wt%, the replacement amount of the feed part in the control group by functional forage is zero, the replacement amount of functional forage in the low replacement group is 8.47 wt%, and the replacement amount of functional forage in the high replacement group is 16.98 wt%.

[0118] Table 5: Composition ratio of commercial TMR formula and different replacement amounts of functional forage.

[0119]

[0120] This experimental example lasted for four weeks (28 days in total). The cows were fed functional forage manufactured in the same batch every 14 days, and one independently packaged functional forage was opened every day; in other words, during the 28 days of this experiment, the cows were fed functional forage manufactured in the first batch and subjected to a complete second-stage anaerobic fermentation for 14 to 28 days from the 1st to the 14th day, and functional forage manufactured in the second batch and subjected to a complete second-stage anaerobic fermentation for 14 to 28 days from the 15th to the 28th day. More specifically, during the experiment, the test sample fed to the cows on the 1st day of every 14 days was functional forage that had undergone a complete second-stage anaerobic fermentation for 14 days (i.e., the product of Example 2 of the present invention), and so on. The functional forage fed on the 2nd to the 14th day of every 14 days was sequentially fed functional forage that had undergone a complete second-stage anaerobic fermentation for 15 to 28 days; therefore, the functional forage fed on the 7th day of every 14 days was the product of Example 3 of the present invention (the second-stage fermentation reached 21 days), and the functional forage fed on the 14th day of every 14 days was the product of Example 4 of the present invention (the second-stage fermentation reached 28 days).

[0121] 〔Measurement process〕

[0122] The cows in this experimental example were raised in a livestock house equipped with cattle beds and milked twice a day, at 4:30 am and 4:30 pm respectively. The measurement process for the somatic cell count in the milk collected in this experimental example is briefly described as follows:

[0123] (1) Before milking, the cow's nipples must be cleaned and dried to avoid foreign objects from entering the milk. The middle milk is collected before and after milking. Do not sample milk from clinically mastitic cows. Since the somatic cell count in the milk components of the fore, middle, and hind milking is very different, it is necessary to collect the middle milk to be representative.

[0124] (2) The test cows collected milk samples continuously twice in the morning and afternoon on the day before the start of the experiment (Day 0), the 14th day, and the 28th day of the experiment, and 25 ml of milk samples were collected in test bottles respectively.

[0125] (3) Mix the morning and afternoon milk samples until the test bottle is about eight-tenths full.

[0126] (4) Immediately cap the bottle after filling it with the milk sample and refrigerate it at about 4°C.

[0127] (5) Place the milk sample in an ice bucket with sufficient frozen ice bars and send it to the Dairy Herd Improvement (DHI) milk sample testing center. Use a somatic cell analyzer to analyze and test the content of somatic cells. The results are shown in Table 6 below.

[0128] Table 6: Test results of somatic cell counts in milk samples collected from cows after feeding different functional forages to replace the TMR formula.

[0129]

[0130] The results of this test example regarding the determination of the improvement effect of functional forage on bovine mastitis are shown in Table 6 above. As can be seen from the above, before the test, the somatic cell count in the milk sample of the control group was the lowest, followed by the low replacement group, and the highest in the high replacement group; by the 14th day of the test, the somatic cell count in the milk sample of the control group increased instead, but the somatic cell counts in the milk samples of the low replacement group and high replacement group of functional forage decreased significantly. In particular, the somatic cell amount in the milk sample of the high replacement group decreased significantly to less than 50% of that before the test (before feeding the functional forage of the present invention). This shows that the functional forage prepared by the two-stage fermentation of the present invention has a significant effect on improving bovine mastitis compared with the commercial TMR formula diet.

[0131] Test Example 5: Determination of the milk production increasing effect of functional forage and the milk quality of the increased milk volume

[0132] The method for determining the milk production increasing effect of the functional forage and the milk quality of the increased milk volume in the foregoing examples is as follows, and the results are shown in Table 7, Table 8 and Figure 2 as shown below.

[0133] The determination of the milk production of cows in this test example is based on the experimental animals and experimental diets in Test Example 4. During the test period of Test Example 4, milking was performed at 5:00 am and 3:30 pm every day (i.e., milking twice a day), and the milk production of individual cows was recorded by the milking system to evaluate the change in milk production of cows before and after feeding different formula diets. The milk production of cows in this test example is statistically analyzed according to the milk production of three groups (control group, low replacement group, high replacement group) in 28 days and referring to the statistical curve of lactation days; in addition, based on the fact that mastitis occurred in cows during the test period resulting in invalid data, only 6 cows with valid data were selected from 7 cows in each group to analyze the change in milk production; the determination results are shown in Table 7 below.

[0134] Table 7: Shows Figure 2Data on the changes in milk production of three groups of cows (control group, low substitution group, high substitution group) within four weeks.

[0135]

[0136] The results of this test example on the determination of the effect of functional forage on increasing milk production of cows are as shown in Table 7 above and Figure 2 as follows. It can be clearly seen from Figure 2 that most of the cows in the control group showed a decrease in milk production, and the maximum decrease was more than 5 kg, while the increase in milk production did not reach 3 kg; the increase in milk production of cows in the low substitution group was very significant, and all the cows in this group had an increase in milk production, with the minimum increase being higher than 1 kg and the maximum increase being equivalent to 3 kg; although the milk production of cows in the high substitution group decreased, the cow with the highest increase in milk production significantly exceeded 4 kg, which was the highest milk production among all cows.

[0137] As can be seen from Table 7, the total change in milk production of cows C1 to C6 in the control group over four weeks was a decrease of 6.31 kg, and the average value of the four-week average milk production change in the control group was a decrease of 1.05 kg; the total change in milk production of cows L1 to L6 in the low substitution group over four weeks was an increase of 15.44 kg, and the average value of the four-week average milk production change in the low substitution group was an increase of 2.57 kg; the total change in milk production of cows H1 to H6 in the high substitution group over four weeks was a decrease of 1.1 kg, and the average value of the four-week average milk production change in the high substitution group was a decrease of 0.18 kg. It can be seen from the above that the use of the functional forage of the present invention is helpful for the milk production that gradually decreases with the number of lactation days after the lactation peak.

[0138] To illustrate the effect of the functional forage of the present invention on increasing milk production with the same standard, the average value (M) of the four-week average milk production change in the control group was used as the basis for normalization. The normalization formula is: Normalized base value = (Average value of other groups - Average value of the control group). Based on the above, the normalized value of the four-week average milk production change in the low substitution group was 3.62 kg, and the normalized value of the four-week average milk production change in the high substitution group was 0.87 kg; it can be seen from the above that the low substitution group had an increase in milk production of 3.62 kg / 4 weeks compared to the control group, and the high substitution group had an increase in milk production of 0.87 kg / 4 weeks compared to the control group, indicating that compared with the general commercial TMR formula feed (control group), regardless of whether the substitution amount of the functional forage of the present invention in the TMR formula feed is high or low, it has a significant effect on increasing the milk production of cows.

[0139] The milk quality determination of the cows in this test example was based on the experimental animals and experimental diets in Test Example 4, to prove that while the cows fed with the forage containing the functional forage of the present invention increased the milk yield, they also maintained the milk quality. By collecting milk samples of the individual cows participating in the test every 14 days during the test period of Test Example 4 (at 4:30 am and pm), after mixing the morning and afternoon milk samples collected on the same day for each individual cow every 14 days, they were sent to the DHI milk sample inspection center for analysis of milk quality detection items such as milk fat rate, lactose rate, milk protein rate, fat-free solids rate, total solids rate, and milk urea nitrogen. The analysis results are shown in Table 8 below.

[0140] Table 8: Detection results of milk fat rate, lactose rate, milk protein rate, fat-free solids rate, total solids rate, and milk urea nitrogen in milk samples collected after cows were fed different functional forages to replace the TMR formula.

[0141]

[0142] As can be seen from Table 8, in the measurements on Day 0, Day 14, and Day 28, the milk fat percentages of the low replacement group and the high replacement group were both higher than those of the control group. Especially on Day 0 and Day 28, the milk fat percentages of the high replacement group reached 4.05% and 3.85% respectively, which were higher than 3.52% and 3.61% of the control group. Among the three measurement time points, the milk protein percentages of the low replacement group and the high replacement group were slightly higher than or close to the values of the control group. On Day 14 and Day 28, the lactose percentage of the high replacement group was slightly lower than that of the control group; but the difference was not significant, indicating that the lactose content did not fluctuate greatly due to the increased replacement. Among the three measurement time points, the fat-free solids percentages of the low replacement group and the high replacement group were similar to those of the control group. At all three measurement time points, the total solids rates of the low replacement group and the high replacement group were higher than those of the control group. Especially on Day 0 and Day 28, the high replacement group reached 12.76% and 12.39% respectively, which were significantly higher than 12.21% and 12.32% of the control group. Among the three measurement time points, the milk urea nitrogen of the low replacement group was higher than that of the control group, while the high replacement group was slightly lower than the control group. Overall, the differences among the three groups were not significant. Based on the above analysis, it can be seen that the low replacement group and the high replacement group both had good performances in milk fat rate, milk protein rate, and total solids rate, which are all important indicators of milk quality. Especially the total solids rate, which is an important indicator for evaluating the nutritional value and quality of milk, and both the low replacement group and the high replacement group were better than the control group. Therefore, from the above data, it can be seen that the low replacement group and the high replacement group not only increased the milk volume, but also successfully maintained the high-quality milk quality.

[0143] In summary, the functional forage obtained by the two-stage fermentation preparation method of first aerobic and then anaerobic fermentation of the present invention has the characteristics of short fermentation time, low protein fermentation loss, fast acid production, good stability, and low ammonia concentration. Moreover, the functional forage of the present invention has a high dry matter content (which helps to increase the daily dry matter intake and the milk production potential of animals), contains short-chain fatty acids such as lactic acid, acetic acid, and butyric acid that contribute to fermentation stability and preservation. Additionally, the rumen-degradable protein content is also higher than that of corn silage. The functional forage of the present invention can maintain the stability of the functional forage after opening due to the decomposition of the remaining sugars during the second-stage anaerobic fermentation, and is durable and easy to store, and the stable components are conducive to the stable utilization of the intestinal microorganisms of herbivores. In addition, when the functional forage of the present invention is used to prepare feed products, it has better technical effects in increasing the milk production of herbivores, maintaining the milk quality, and improving the mastitis of herbivores compared with ordinary corn silage.

[0144] The embodiments described above are only used to illustrate the technical ideas and characteristics of the present invention, and the purpose is to enable those skilled in the art to understand the content of the present invention and implement it accordingly, rather than to limit the patent scope of the present invention. That is, equivalent changes or modifications made generally in accordance with the spirit disclosed in the present invention should still be covered within the patent scope of the present invention.

[0145] References:

[0146] [1] Huisden CM, Adesogan AT, Kim SC and Ososanya T, Effect of applying molasses or inoculants containing homofermentative or heterofermentative bacteria at two rates on the fermentation and aerobic stability of corn silage. J Dairy Sci 92:690–697(2009).

[0147] [2] Van Soest PJ, Robertson JB and Lewis BA, Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J Dairy Sci 74:3583–3597(1991).

[0148] [3]Morris DL,Quantitative determination of carbohydrates with Drey-wood's anthrone reagent.Science 107:254–255(1948).

[0149] [4]Danner H,Madzingaidzo L,Holzer M,Mayrhuber L and Braun R,Extrac-tion and purification of lactic acid from silages.Bioresour Technol 75:181–187(2000).

[0150] [5]Weatherburn MW,Phenol-hypochlorite reaction for determination ofammonia.Anal Chem 39:971–974(1967).

[0151] [6]Ashbell G,Weinberg ZG,Azrieli A,Hen Y and Horev B,A simple systemto study the aerobic deterioration of silages.Can Agric Eng 33:391–393(1991).

[0152] [7]Hu W,Schmidt RJ,McDonell EE,Klingerman CM and Kung L Jr,The effectof Lactobacillus buchneri 40788or Lactobacillus plantarum MTD-1on thefermentation and aerobic stability of corn silages ensiled at two dry mattercontents.J Dairy Sci 92:3907–3914(2009).

[0153] [8]Nocek, J.E. 1985. Evaluation of specific variables affecting in situ estimates of ruminal dry matter and protein digestion. J. Anim. Sci. 60: 1347 - 1358.

[0154] [9]Nutrient Requirements of Dairy Cattle, Seventh Revised Edition, 2001 (2001) p292 Table 15 - 2a.

Claims

1. A functional forage grass, which comprises a raw material that contains a forage matrix material and a nutrient material. The raw material is made into the functional forage grass after undergoing a first-stage aerobic fermentation with fungi and a second-stage anaerobic fermentation with lactic acid bacteria and yeast.

2. The functional forage grass according to claim 1, wherein, Calculated based on the total dry matter content of the raw material being 100 wt%, the raw material includes 40 wt% to 60 wt% of forage grass, 10 wt% to 20 wt% of soybean meal, and 30 wt% to 40 wt% of wheat bran.

3. The functional forage grass according to claim 1, wherein, The functional forage grass contains a total organic acid content of up to 160 grams per kilogram.

4. The functional forage grass according to claim 1, wherein, The functional forage grass contains up to 210 grams of lactic acid (Lactic acid, g / kg) per kilogram.

5. The functional forage grass according to claim 1, wherein, The functional forage grass contains up to 74 grams of acetic acid (Acetic acid, g / kg) per kilogram.

6. The functional forage grass according to claim 1, wherein, The functional forage grass contains up to 40 grams of butyric acid (Butyric acid, g / kg) per kilogram.

7. The functional forage grass according to claim 1, wherein The dry matter content of the functional forage grass is up to 42 wt%.

8. The functional forage grass according to claim 1, wherein, The rumen crude protein digestibility of the functional forage grass reaches 16%.

9. The functional forage grass according to claim 1, wherein, Calculated based on the total protein content of the functional forage grass, the rumen-degraded protein content of the functional forage grass reaches 77% of the total protein.

10. A method for preparing the functional forage grass according to any one of claims 1 to 9, which includes a first-stage aerobic fermentation and a second-stage anaerobic fermentation, wherein, The steps of the first-stage aerobic fermentation include: Cut the forage grass into a forage matrix material; Add the nutrient material and uniformly mix it with the forage matrix material and adjust the moisture content to form a mixed material; After sterilizing the mixed material, add fungi to the mixed material; After the mixed material is colonized by the fungi, place it in an aerobic and constant-temperature environment for aerobic fermentation for 1 to 2 months to obtain a first fermentation product; The steps of the second-stage anaerobic fermentation include: Add yeast and lactic acid bacteria to the first fermentation product, then mix and stir and adjust the moisture content to form a forage material; Pack the forage material to form an anaerobic fermentation environment, and carry out anaerobic fermentation for 7 to 28 days to obtain the functional forage grass.

11. The preparation method of the functional forage according to claim 10, wherein, The forage matrix material can be selected from one or any combination of Pennisetum alopecuroides, Napier grass, Sorghum sudanense, Sweet oats, Wheatgrass, Trifolium alexandrinum.

12. The preparation method of the functional forage grass according to claim 10, wherein, The nutrient material is selected from one or any combination of soybean meal, wheat bran, and corn cob.

13. The preparation method of the functional forage grass according to claim 10, wherein, In the first-stage aerobic fermentation, calculated based on the weight of the mixed material, the addition amount of the fungi is 0.1 wt% to 10 wt%; the fungi are one or any combination of edible fungi and medicinal fungi.

14. The preparation method of the functional forage according to claim 10, wherein, In the second-stage anaerobic fermentation, calculated based on the weight of the first fermentation product, the addition amount of the yeast is 0.1 wt% to 10 wt%, and the addition amount of the lactic acid bacteria is 0.1 wt% to 10 wt%.

15. The preparation method of the functional forage grass according to claim 10, wherein, In the first-stage aerobic fermentation, calculated based on the total weight of the raw material, the total moisture content of the raw material is 45 wt% to 65 wt%, and the total dry matter content of the raw material is 35 wt% to 55 wt%.

16. The preparation method of the functional forage according to claim 15, wherein, The nutritional material is soybean powder and wheat bran; calculated based on the total dry matter content of the raw materials being 100 wt%, 40 wt% to 60 wt% of the total dry matter content of the raw materials comes from forage grass, 10 wt% to 20 wt% comes from soybean powder, and 30 wt% to 40 wt% comes from wheat bran.

17. Use of the functional forage grass according to any one of claims 1 to 9 for preparing a feed product for improving mastitis in herbivores.

18. Use of the functional forage grass according to any one of claims 1 to 9 for preparing a feed product for increasing the milk production of herbivores.

19. Use of the functional forage grass prepared by the preparation method according to any one of claims 10 to 16 for preparing a feed product for improving mastitis in herbivores.

20. Use of the functional forage grass prepared by the preparation method according to any one of claims 10 to 16 for preparing a feed product for increasing the milk production of herbivores.

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