Alfalfa silage containing artemisia sieversiana and preparation method thereof

Abstract

The invention belongs to the technical field of feed, and particularly discloses alfalfa silage containing artemisia sieversiana and a preparation method thereof.The preparation method comprises the steps that fresh alfalfa with the water content being 60%-80% in the squaring stage is taken and then cut up, and an alfalfa silage raw material is obtained; the method comprises the following steps: adding 0.625-5% of artemisia sieversiana powder into an alfalfa silage raw material, uniformly mixing, and carrying out silage fermentation at 15-37 DEG C for 30-90 days to obtain the alfalfa silage containing the artemisia sieversiana. Compared with the prior art, the silage additive has the advantages that the artemisia sieversiana is used as the silage additive to be jointly fermented with the silage raw materials, the fermentation quality of the silage can be improved, and compared with the silage without increasing the size of the artemisia sieversiana, the prepared silage has the advantages that the pH value is reduced, and / or the dry matter content is increased, and the beneficial bacterium content is increased.

Description

Technical Field

[0001] The invention relates to the technical field of feed, in particular to alfalfa silage containing Artemisia macrophylla and a preparation method thereof. Background Art

[0002] Alfalfa is a perennial leguminous forage grass with a high protein content and rich in vitamins and minerals. It is an important forage resource. Silage is an effective and economical measure to preserve alfalfa green fodder, which can solve the problem that alfalfa hay is not suitable for preparation in the rainy season. However, it is difficult to obtain high-quality green fodder from alfalfa in the natural state because of its low sugar content and high buffering energy value. The use of additives can speed up the fermentation of silage and improve the quality of alfalfa silage. At present, the additives used in silage mainly include lactic acid bacteria agents and organic acids. These additives improve the quality of alfalfa silage by inhibiting the activity of microorganisms, thereby meeting the requirements of ruminants for feed. However, many additive products have good effects but cannot be widely used due to their high cost. The development of new additives, especially low-cost, green and pollution-free additives, is an important development direction of silage additives. Artemisia grandis contains rich nutrients, including crude protein, amino acids and mineral elements. Studies have shown that the crude protein content of Artemisia grandis is high, reaching 3.34%, and the total amino acid content is 17.59%, which is the most outstanding among the three Artemisia plants. In addition, Artemisia selengensis also contains a variety of mineral elements, such as potassium and iron, which can provide comprehensive nutritional support for animals. Artemisia selengensis can be used as a feed additive for livestock, especially in seasons when forage is scarce. Its young branches and inflorescences can be made into hay as a reserve feed for livestock in winter. This application can not only improve the nutritional value of feed, but also improve the growth performance and health of animals to a certain extent.

[0003] If Artemisia annua can be used as silage, it can improve the growth performance of animals and enhance their disease resistance. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides a kind of alfalfa silage containing Artemisia macrophylla and a preparation method thereof.

[0005] To achieve the above object, the present invention is implemented according to the following technical solutions:

[0006] One of the purposes of the present invention is to provide an alfalfa silage containing Artemisia grandis, wherein 0.625% to 5% of Artemisia grandis powder based on the mass of fresh alfalfa grass, a raw material of the alfalfa silage, is added to the alfalfa silage; the Artemisia grandis powder is powder obtained by crushing the whole plant of Artemisia grandis through a 10-120 mesh sieve.

[0007] Furthermore, the water content of the fresh alfalfa grass as the raw material of the alfalfa silage is 50% to 80%; the water content of the Artemisia selengensis powder is 5% to 20%.

[0008] The second object of the present invention is to provide a method for preparing alfalfa silage containing Artemisia grandis, comprising the following steps:

[0009] S1, taking fresh alfalfa grass at the budding stage with a moisture content of 60%-80%, and then chopping it to obtain alfalfa silage raw material;

[0010] S2. Add 0.625% to 5% of Artemisia annua powder to the alfalfa silage raw material, mix well, and then ferment at 15-37° C. for 30-90 days to obtain alfalfa silage containing Artemisia annua.

[0011] Compared with the prior art, the present invention uses Artemisia selengensis as a silage additive and ferments it together with silage raw materials, which can improve the fermentation quality of silage. Compared with the control without Artemisia selengensis, the prepared silage has a lower pH value and / or an increased dry matter content and an increased content of beneficial bacteria. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 This is the effect of Artemisia annua on alfalfa silage microorganisms (species level). DETAILED DESCRIPTION

[0013] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with embodiments. The specific embodiments described herein are only used to explain the present invention and are not used to limit the invention.

[0014] 1. Preparation of alfalfa silage

[0015] Adding Artemisia grandis to alfalfa at a ratio of 0.625%, 1.25%, 2.5% and 5% of fresh alfalfa grass for silage preparation, the specific method is as follows:

[0016] Fresh alfalfa grass (water content of 71.81%) in the budding stage was taken, and then the grass was cut into 3-5 cm lengths on a chopper to obtain silage raw materials. The silage raw materials were divided into three groups, namely FA1 (0.625% of Artemisia selengensis), FA2 (1.25% of Artemisia selengensis), FA3 (2.5% of Artemisia selengensis), FA4 (5% of Artemisia selengensis) and a control group FC. The amounts of silage additives added to the above three groups were as follows: FA1 was added according to the standard of adding 0.0625 kg of Artemisia selengensis powder to 10 kg of fresh grass (water content of 72%); FA2 was added according to the standard of adding 0.125 kg of Artemisia selengensis powder to 10 kg of fresh grass (water content of 72%); FA3 was added according to the standard of adding 0.25 kg of Artemisia selengensis powder to 10 kg of fresh grass (water content of 72%); FA4 was added according to the standard of adding 0.5 kg of Artemisia selengensis powder to 10 kg of fresh grass (water content of 72%); and the control group did not add any substance.

[0017] Then, the Artemisia selengensis powder was mixed into fresh alfalfa grass (water content 72%) according to the above-mentioned addition amount, and the fresh grass was put into a polypropylene bag, sealed with a vacuum sealer, and silage fermented at 15-37° C. After 45 days of silage, the bags were opened and samples were taken for laboratory analysis, mainly for pH value, dry matter and crude protein content.

[0018] The various analysis methods are as follows:

[0019] 1) Determination of dry matter

[0020] Dry matter: The remaining part after removing the free water and bound water of the plant is the dry matter of the plant. The test method adopts the oven drying method. The sample is weighed and the weight is recorded, and then dried in an oven at 65℃ for 48 hours until constant weight is reached, and the weighed weight is recorded to calculate the dry matter (DM) content.

[0021] 2) Determination of pH value

[0022] After opening the silage, 20 g of sample was weighed, 180 mL of distilled water was added, stirred evenly, and stirred for 1 min using a juicer. The mixture was filtered through four layers of gauze and filter paper, and the pH value of the filtrate was measured using a pH meter (Raymag S-3C Precision Meter, Shanghai Precision Scientific Instrument Co., Ltd.).

[0023] 3) Determination of organic acids

[0024] Lactic acid and volatile fatty acids (acetic acid, propionic acid and butyric acid) were determined by high performance liquid chromatography SHIMADZE-10A. Chromatographic column: Shodex RspakKC-811S-DVB gel Column 30×8mm; detector: SPD-M10AVp; mobile phase: 3mmol / L perchloric acid; flow rate: 1 mL / min; column temperature: 50℃; detection wavelength: 210nm; injection volume: 5μL.

[0025] 4) Determination of crude protein (CP)

[0026] Use the Kjeldahl method to determine nitrogen. Weigh 0.5000g of dried sample and put it into a digestion tube. Add a mixture of potassium sulfate and copper sulfate as a catalyst. The mixing ratio of potassium sulfate and copper sulfate is 10:1. Add 10ml of concentrated sulfuric acid and digest at 420℃. Digest until the color changes to clear blue. Prepare 40% NaOH, 1% boric acid absorption solution and 0.1M hydrochloric acid solution, and titrate and calibrate the 0.1M hydrochloric acid solution. Use a fully automatic nitrogen analyzer (FOSS fully automatic Kjeldahl nitrogen analyzer) for determination.

[0027] The experimental steps are as follows: 1. Sample weighing: weigh about 0.5g of dry sample and transfer all of it into the digestion tube 2. Digestion: add 10ml of concentrated sulfuric acid and about 3g of CuSO4 to the digestion tube 4 .5H 2 0 and K 2 SO 4 Mixed catalyst (whose mass ratio is CuSO 4 .5H 2 0:K 2 SO 4 =1:10), put it into a digestion furnace and digest it at 420℃ for about 1 hour until the sample becomes light green and transparent; 3. Use Kjeldahl nitrogen analyzer to measure.

[0028] 5) Analysis of microorganisms

[0029] Alfalfa silage samples were taken to extract microbial genomic DNA, and the extracted genomic DNA was detected by 1% agarose gel electrophoresis. The qualified DNA samples were randomly broken into fragments of about 350 bp in length using a Covaris ultrasonic crusher, and the library preparation was completed by end repair, A-tailing, sequencing adapter addition, purification PCR amplification and other steps. After the library construction was completed, Qubit2.0 was used for preliminary quantification, and then the insert size of the library was detected by Agilent2100. After the insert size met the expectations, the effective concentration of the library was accurately quantified by the Q-PCR method (library effective concentration>3nM) to ensure the quality of the library. After the library was qualified, NBSEQ-T7RSReagentKit (FCLPE150) version3.0 was used for sequencing on the DNBSEQ-T7 platform. Data analysis started with the original sequence of the machine, and the original sequence was first optimized by splitting, quality shearing and removing contamination. Then the optimized sequence was used for splicing assembly and gene prediction, and the obtained genes were annotated and classified in terms of species and function.

[0030] The data were analyzed using SPSS 17.0 software to comprehensively evaluate the effects of Artemisia grandis on silage fermentation quality, nutrients and microbial distribution. Figure 1 shown.

[0031] Table 1 Effects of Artemisia argyi on alfalfa silage quality

[0032] project FC FA1 FA2 FA3 FA4 pH <![CDATA[5.73 a ±0.01]]> <![CDATA[5.67 ab ±0.02]]> <![CDATA[5.58 b ±0.07]]> <![CDATA[5.46 c ±0.02]]> <![CDATA[5.23 d ±0.02]]> Dry matter (%) <![CDATA[29.62 b ±0.58]]> <![CDATA[30.19 ab ±1.15]]> <![CDATA[30.67 ab ±1.02]]> <![CDATA[32.07 ab ±0.72]]> <![CDATA[33.17 a ±1.04]]> Crude protein (%DM) 16.51±0.29 16.48±0.51 16.27±0.21 17.25±1.42 15.90±0.42 Neutral detergent fiber (%DM) 35.90±0.88 34.56±0.78 35.32±1.17 36.64±0.99 34.82±0.71 Acid detergent fiber (% DM) 29.90±0.19 29.01±0.87 29.10±0.62 29.28±0.81 28.27±0.43 Soluble carbohydrates (%DM) 1.00±0.04 0.98±0.04 0.98±0.01 0.96±0.01 0.98±0.03 Lactic acid (%DM) 5.74±0.13 6.34±0.04 4.69±0.56 5.94±0.66 5.38±0.78 Acetic acid (%DM) 1.84±0.20 2.41±0.27 1.80±0.11 2.34±0.02 1.94±0.06 Propionic acid (%DM) 1.34±0.21 1.50±0.22 1.37±0.27 1.18±0.14 1.24±0.18 Butyrate (%DM) / / / / /

[0033] Note: / indicates not detected. The letters after the numerical values ​​in each row indicate the significance of the difference. There is no significant difference between treatments with at least one identical letter at the 0.05 level, and there is a significant difference between treatments without the same letter at the 0.05 level.

[0034] As shown in the results in Table 1, the pH value of the silage with the addition of Artemisia argyi was lower than that of the control group, among which the pH values ​​of the FA2, FA3 and FA4 treatment groups decreased most significantly. Artemisia argyi had no significant effect on the lactic acid, acetic acid and propionic acid contents of alfalfa silage (P>0.05), and no butyric acid was detected in all treatment groups. The dry matter content of the Artemisia argyi additive treatment group was significantly higher than that of the control group (P<0.05). Artemisia argyi had no significant effect on the content of neutral detergent fiber, acid detergent fiber, crude protein and soluble sugar (P>0.05). The above results show that the addition of Artemisia argyi has a very good effect on maintaining the pH of alfalfa silage in an environment that inhibits the growth of miscellaneous bacteria.

[0035] Depend on Figure 1 The results showed that at the species level, the high-abundance bacteria in alfalfa silage included Latilactobacillus curvatus, Lactiplantibacillusplantarum, Lactiplantibacillussp., Levilactobacillus brevis, Pediococcus pentosaceus, Latilactobacillus sakei, Enterococcus sp., and Lactiplantibacillusparaplantarum. With the increase in the proportion of Artemisia grandis, the relative abundance of Lactiplantibacillusplantarum and Lactiplantibacillus sp. increased. The addition of Artemisia grandis may promote the growth of Lactiplantibacillusplantarum and Lactiplantibacillus sp.

[0036] In summary, the present invention uses Artemisia selengensis as a silage additive and ferments it together with silage raw materials, which can improve the fermentation quality of silage. Compared with the control without adding Artemisia selengensis, the prepared silage has a lower pH value and / or an increased dry matter content, a lower ratio of ammonia nitrogen to total nitrogen, and a lower number of enterobacteria and yeast.

[0037] The technical solution of the present invention is not limited to the above-mentioned specific embodiments. All technical variations made according to the technical solution of the present invention fall within the protection scope of the present invention.

Claims

1. An alfalfa silage containing Artemisia grandis, characterized in that: The alfalfa silage contains 0.625% to 5% of the mass of fresh alfalfa grass, which is the raw material of the alfalfa silage, Artemisia selengensis powder; the Artemisia selengensis powder is powder obtained by crushing the whole plant of Artemisia selengensis through a 10-120 mesh sieve.

2. The alfalfa silage containing Artemisia grandis according to claim 1, characterized in that: The water content of the fresh alfalfa grass, which is the raw material of the alfalfa silage, is 50% to 80%; the water content of the Artemisia selengensis powder is 5% to 20%.

3. A method for preparing alfalfa silage containing Artemisia grandis, characterized in that: The following steps are involved: S1, taking fresh alfalfa grass at the budding stage with a moisture content of 60%-80%, and then chopping it to obtain alfalfa silage raw material; S2. Add 0.625% to 5% of Artemisia annua powder to the alfalfa silage raw material, mix well, and perform silage fermentation at 15-37° C. for 30-90 days to obtain alfalfa silage containing Artemisia annua.

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