Method for improving digestibility of alfalfa silage in saline-alkali soil

Abstract

The invention provides a method for improving digestibility of alfalfa silage in saline-alkali soil. The method comprises the following steps: (1) dressing organic fertilizer and biopesticide in alfalfa production; and (2) chopping the harvested alfalfa raw material, controlling the moisture content, and inoculating a lactobacillus paracasei and / or lactobacillus plantarum inoculant for ensiling to obtain the alfalfa silage with improved digestibility. According to the method, the digestibility and the conversion rate of the alfalfa silage in the saline-alkali soil are improved mainly through a field management technology and subsequent silage additive optimization. Firstly, the quality of the alfalfa in the saline-alkali soil is improved through the cooperative use of the organic fertilizer and the biopesticide, and then the fiber digestibility is improved through the doses of the organic fertilizer and the biopesticide with different concentrations, so that the digestibility of the alfalfa silage is improved.

Description

Technical Field

[0001] This invention relates to the field of silage, and more specifically, to a method for improving the digestibility of alfalfa silage in saline-alkali land. Background Technology

[0002] Saline-alkali land is an underutilized land resource in my country, and alfalfa grown on it typically suffers from low yields and low digestibility. Currently, improving the digestibility of silage mainly relies on additives, and the technical methods employed are relatively limited. To improve the digestibility of alfalfa grown on saline-alkali land, there is an urgent need to systematically research and develop methods that can effectively enhance its digestibility. Summary of the Invention

[0003] The purpose of this invention is to provide a method for improving the digestibility of alfalfa silage in saline-alkali land.

[0004] Topdressing and pesticide application are important field management measures in alfalfa production. However, the optimal application and management schemes for specific types and dosages of fertilizers and pesticides are still unclear and require further in-depth research to obtain more accurate conclusions. This invention determines the optimal fertilizer and pesticide application and management techniques by applying different fertilizer treatments and spraying different pesticide treatments, and measuring and analyzing the nutritional quality of alfalfa from different crops.

[0005] To achieve the objective of this invention, in a first aspect, this invention provides a method for improving the digestibility of alfalfa silage in saline-alkali land, the method comprising: (1) Apply organic fertilizer and biological pesticides during the production of alfalfa (preferably alfalfa grown in saline-alkali land, more preferably alfalfa harvested after the first and second cuts after topdressing, and even more preferably harvested at the budding stage); (2) Chop the harvested alfalfa raw material, control the moisture content, and inoculate with Lactobacillus plantarum (Lactobacillus plantarum). Lactobacillus plants Silage is produced by using microbial agents to obtain alfalfa silage with improved digestibility.

[0006] The biological pesticide mentioned in step (1) is a Beauveria bassiana biological insecticide wettable powder, the active ingredient of which is Beauveria bassiana (Beauveria bassiana). Beauveria bassiana ).

[0007] Further, the organic fertilizer in step (1) has the following composition: organic matter ≥30%, N+P2O5+K2O ≥4% (preferably N+P2O5+K2O ≥5%), wherein the mass ratio of N:P2O5:K2O is 1:1:2; wherein % is the mass percentage content; the pH of the organic fertilizer is ≤8.5; Preferably, the organic matter content is ≥50%.

[0008] Furthermore, the application rate of organic fertilizer is 3000 kg / ha.

[0009] Furthermore, in step (1), the bacterial content of Beauveria bassiana in the biopesticide is 15 billion spores / g; Preferred insecticide wettable powder of Beauveria bassiana (e.g., purchased from Ningxia Zhongwei Taike Biotechnology Co., Ltd., product number PD20141407) is used.

[0010] Furthermore, the application rate of the biopesticide is 750-1500 g / ha, preferably 1125 g / ha.

[0011] Furthermore, the method of applying organic fertilizer and biological pesticide in step (1) is as follows: organic fertilizer is mainly applied at the beginning of the second crop or subsequent crops each year, and additional fertilizer is applied after every two harvests; The application of biological pesticides is mainly based on the local pest occurrence patterns, and is carried out during the larval stage or when harmful insects are found.

[0012] Further, step (2) includes: chopping the harvested alfalfa raw material, controlling the moisture content to 50-70%, and inoculating it with Lactobacillus paracasei and / or Lactobacillus plantarum inoculum (bacterial content of 10). 5 -10 6 Alfalfa silage is obtained by ensiling alfalfa (cfu / g) at 0-40℃ for 30-360 days.

[0013] Preferably, the *Lactobacillus plantarum* is... Lactobacillus plantarum Wuzhe25, this strain is now deposited at the China General Microbiological Culture Collection Center, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, 100101, China, accession number CGMCC No. 33705, deposit date March 3, 2025.

[0014] Secondly, the present invention provides alfalfa silage prepared by the method.

[0015] Thirdly, the present invention provides the application of the alfalfa silage in animal husbandry.

[0016] The animals mentioned include, but are not limited to, cattle and sheep.

[0017] By employing the above technical solution, the present invention has at least the following advantages and beneficial effects: This invention primarily improves the digestibility and conversion rate of alfalfa silage in saline-alkali land through field management techniques and subsequent optimization of silage additives. Firstly, the combined use of organic fertilizer and biopesticides improves the quality of alfalfa in saline-alkali land. Then, different concentrations of organic fertilizer and biopesticide dosages increase fiber digestibility, thereby improving the digestibility of alfalfa silage. Alfalfa in saline-alkali land grows slowly and has a high lignocellulose content, typically resulting in low digestibility. To improve the digestibility of alfalfa silage in saline-alkali land, lactic acid bacteria additives are commonly used, but the improvement effect is limited.

[0018] This invention employs multiple measures to improve the digestibility of alfalfa silage. These measures include field management practices and silage preparation, which, when used in combination, significantly enhance the digestibility of alfalfa silage, increasing it by more than 10% compared to methods without any topdressing or using chemical fertilizers and pesticides. Furthermore, the improvement in digestibility remains significant in subsequent crops. In contrast, the effects of topdressing with chemical pesticides and fertilizers diminish in subsequent crops, and even with additives, the improvement is minimal. Therefore, applying organic fertilizers and biological pesticides not only reduces the number of operations but also effectively improves the digestibility of alfalfa products in subsequent crops, achieving a reduced workload while providing a long-term increase in digestibility. Detailed Implementation

[0019] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0020] The *Lactobacillus plantarum* LP1 (accession number CICC 6240) and *Lactobacillus paracasei* LP2 (accession number CICC 6263) used in the following examples were purchased from the China Center of Industrial Culture Collection (CICC).

[0021] Example 1 The alfalfa experimental field used in the experiment was located at the Guying Base of Zhengzhou Tianyuan Muge Grass Industry Co., Ltd., while the indoor experiment was conducted at China Agricultural University. The alfalfa used in the experiment was variety WL525HQ. Organic fertilizer (Tuxiucai fish protein high-calcium bio-fertilizer, purchased from Kaifeng Jiarun Agricultural Technology Co., Ltd.), high-sulfur nitrogen fertilizer (purchased from Kaifeng Jiarun Agricultural Technology Co., Ltd.), biological pesticides (purchased from Ningxia Zhongwei Taike Biotechnology Co., Ltd., product number PD20141407), and chemical pesticides (fluoxadixyl, indoxacarb, and Yingerguo orange peel hydrosol) were purchased from Henan Qizhixing Biotechnology Co., Ltd.

[0022] The tested organic fertilizer product specifications are: organic matter ≥50%, N+P2O5+K2O ≥5%, where the mass ratio of N:P2O5:K2O is 1:1:2.

[0023] The tested biological pesticide was a Beauveria bassiana biological insecticide wettable powder. The active ingredient and its content were: Beauveria bassiana. Beauveria bassiana The bacterial content is approximately 15 billion spores per gram.

[0024] The experiment primarily focused on the selection of topdressing and pesticides during alfalfa growth. The experiment included two factors: topdressing treatment and pesticide treatment. Five pesticide treatment groups were included: no pesticide; conventional chemical pesticide; 375 g / ha of biological pesticide; 750 g / ha of biological pesticide; and 1125 g / ha of biological pesticide. The conventional chemical pesticide treatment used a mixture of 5% flufenoxuron, indoxacarb, and the adjuvant, Inergo orange peel hydrosol, with a mass ratio of 6:3:1 and application rates of 900 g / ha, 450 g / ha, and 150 g / ha, respectively. Five fertilizer treatment groups were also included: no fertilizer; chemical fertilizer; 750 kg / ha of organic fertilizer; 1500 kg / ha of organic fertilizer; and 3000 kg / ha of organic fertilizer. The chemical fertilizer treatment used a high-sulfur nitrogen fertilizer at an application rate of 300 kg / ha. Each treatment group had 3 replicates, resulting in a total of 75 plots (5 pesticide treatments × 5 fertilizer treatments × 3 replicates), with each plot measuring 13 m × 14 m. A buffer zone was maintained between each plot to prevent cross-influence. Alfalfa was sown in November 2022, and basal fertilizer (150 kg / ha ammonium sulfate + 105 kg / ha potassium sulfate) was applied in March 2023. Topdressing and pesticide application were carried out on August 19th according to the experimental design. Nutrient components of the plant samples were determined by harvesting at five points on August 29th and November 2nd, at the fifth and sixth budding stages of the alfalfa crop.

[0025] Chop the harvested alfalfa raw material, control the moisture content to about 60%, and inoculate it with Lactobacillus plantarum inoculant LP1 (accession number CICC 6240), Lactobacillus paracasei inoculant LP2 (accession number CICC 6263), or Lactobacillus plantarum inoculant LP25 (i.e., Lactobacillus plantarum Wuzhe25 (CGMCC No. 33705), the bacterial content of the inoculum is 10. 5Alfalfa silage was obtained by ensiling the alfalfa silage for 60 days under outdoor conditions (maximum temperature below 20℃, average temperature about 6℃). Approximately 100 g of the alfalfa silage sample was weighed and placed in an oven at 65℃ for 4 hours to determine its dry matter content (DM). After drying, the sample was pulverized and passed through a 40-mesh sieve. The crude protein (CP) content was determined using the Kjeldahl method. The neutral detergent fiber (NDF) content was determined using the Van Soest method. The crude ash (Ash) content was determined using the muffle furnace combustion method.

[0026] Digestibility assay (in vitro method) 1. Collection of rumen fluid Rumen fluid was collected from four Angus cattle with permanent rumen cannulas from Beijing Doudian Hengsheng Animal Husbandry Technology Co., Ltd., before morning feeding. Rumen contents, including a mixture of rumen fluid and residue, were collected through the rumen cannulas of the cattle. This mixture was filtered through four layers of gauze, and the extracted rumen fluid was quickly poured into a preheated 39°C thermos and immediately transported to the laboratory. All rumen fluid was mixed thoroughly according to the specified ratio and placed in a 39°C constant temperature water bath for later use.

[0027] 2. Determination of in vitro rumen digestibility 0.5 g of pulverized sample, dried at 65℃ and passed through a 10-mesh sieve, was weighed and placed in a filter bag for in vitro digestibility determination. The 48-h in vitro dry matter digestibility (IVDMD) was determined using a DaisyIII incubator, and the 48-h in vitro neutral detergent fiber digestibility (IVNDFD) and 48-h in vitro acid detergent fiber digestibility (IVADFD) were determined using a semi-automatic fiber analyzer (ANKOM220, USA).

[0028] Table 1. Effects of topdressing and pesticide application on the in vitro digestibility of alfalfa feed from the first harvest after fertilization and pesticide application.

[0029] As shown in Table 1, both pesticide and fertilizer treatments had a highly significant impact on the IVDMD and IVNDFD of alfalfa raw materials harvested for the first time after fertilization. P <0.01), there was no significant difference in IVADFD ( P>0.05). Different fertilizer types and dosages have different effects on IVDMD. The treatment with no fertilizer and no pesticides had the lowest IVDMD, while the treatment with no fertilizer but with biopesticides had the highest IVDMD. After applying chemical fertilizers, the IVDMD increased in both chemical and biopesticide treatments, but the treatment with chemical pesticides combined with chemical fertilizers had the highest IVDMD at 62.93%, while the treatment with biopesticides and organic fertilizers reached 63.64%. The highest IVNDFD and IVADFD also appeared in the biopesticide and organic fertilizer treatment groups. Compared with chemical fertilizer and chemical pesticide treatments, biopesticides and organic fertilizers can effectively improve the digestibility of alfalfa. Topdressing with organic fertilizer and using biopesticides can improve the digestibility of alfalfa feedstock. Compared with no topdressing and no pesticides, the increase in dry matter digestibility is approximately 11.55%, and the increase in NDF digestibility is 11.95%. Compared with the application of chemical fertilizers and pesticides, the increase in dry matter digestibility was approximately 2.13%, the increase in NDF digestibility was 5.48%, and the increase in ADF digestibility was 4.99%. Based on the above experimental data, it can be concluded that the application of organic fertilizers and biological pesticides is more effective than the application of chemical fertilizers and pesticides in improving the digestibility of alfalfa.

[0030] Table 2. Effects of topdressing and pesticide application on the in vitro digestibility of alfalfa feed from the second harvest after fertilization and pesticide application.

[0031] Table 2 shows that, under various treatments, the digestibility of alfalfa harvested twice after the second harvest was still the highest after applying 1125 g / ha of biological pesticide and 3000 kg / ha of organic fertilizer. Compared with no pesticide or fertilizer application, the increase in dry matter digestibility was up to 9.55%, ADF digestibility up to 11.74%, and NDF digestibility up to 10.15%. Compared with the application of chemical pesticides and topdressing with chemical fertilizers, the increase in dry matter digestibility was up to 3.14%, ADF digestibility up to 5.20%, and NDF digestibility up to 5.63%. These results indicate that topdressing with organic fertilizer and biological pesticides effectively improved the digestibility of alfalfa harvested twice after fertilization, and the effect was better than that of topdressing with chemical fertilizers and pesticides. The effects of different treatments on the in vitro digestibility of the fifth crop of alfalfa silage are shown in Tables 3-5.

[0032] Table 3. Effects of different treatments on the in vitro digestibility of fifth-crop alfalfa silage

[0033] LP1: CICC 6240. LP2: CICC 6263.

[0034] In vitro dry matter digestibility data showed differences in dry matter digestibility between alfalfa silage obtained from the first harvest after fertilization and pesticide application. The digestibility of alfalfa silage treated with chemical pesticides was lower than that obtained from a combination of chemical pesticides and organic fertilizers. Treatments using 1125 g / ha of biological pesticides and 3000 kg / ha of organic fertilizers achieved higher digestibility compared to other treatments. Compared to other additives and no additives, LP25 significantly improved dry matter digestibility, reaching over 69%, representing a 16.96% increase compared to no fertilization, no pesticides, and no additives.

[0035] Table 4. Effects of different treatments on the in vitro digestibility of the fifth crop of alfalfa silage (continued from Table 3)

[0036] In vitro neutral detergent fiber (NDF) digestibility data showed differences in NDF digestibility between alfalfa silage obtained from the first harvest after fertilization and pesticide application. The digestibility of alfalfa silage treated with chemical pesticides was lower than that obtained from a combination of chemical pesticides and organic fertilizers. Treatments using 1125 g / ha of biological pesticides and 3000 kg / ha of organic fertilizers achieved higher digestibility compared to other treatments. Compared to other additives and no additives, LP25 significantly improved NDF digestibility, reaching over 58%, representing a 15.25% increase compared to no fertilization, no pesticides, and no additives.

[0037] Table 5. Effects of different treatments on the in vitro digestibility of the fifth crop of alfalfa silage (continued from Table 4)

[0038] The in vitro acid detergent fiber digestibility data showed differences in the digestibility of alfalfa silage obtained from the first harvest after fertilization and pesticide application. The digestibility of alfalfa silage treated with chemical pesticides was lower than that obtained from a combination of chemical pesticides and organic fertilizers. Treatments using 1125 g / ha of biological pesticides and 3000 kg / ha of organic fertilizers achieved higher digestibility compared to other treatments. Compared to other additives and no additives, LP25 significantly improved the digestibility of acid detergent fiber, reaching over 46%, and the improvement was 20.96% compared to no fertilization, no pesticides, and no additives. The effects of different treatments on the in vitro digestibility of the sixth crop of alfalfa silage are shown in Tables 6-8.

[0039] Table 6. Effects of different treatments on the in vitro digestibility of sixth-crop alfalfa silage

[0040] Although no fertilization was applied after the fifth alfalfa harvest, the effects of topdressing with organic fertilizer and biopesticides persisted into the sixth harvest. In vitro dry matter digestibility data showed a difference in dry matter digestibility between alfalfa silage obtained from the second harvest after fertilization and pesticide application. The digestibility of the chemical fertilizer treatment with chemical pesticides was lower than that of alfalfa silage obtained from the combined application of chemical pesticides and organic fertilizers. The treatment with 1125 g / ha of biopesticides and 3000 kg / ha of organic fertilizers achieved higher digestibility compared to other treatment groups. Compared to other additives and no additives, LP25 significantly improved the digestibility of acid detergent fiber, reaching over 71%, representing a 14.06% increase compared to no topdressing, no pesticides, and no additives.

[0041] Table 7. Effects of different treatments on the in vitro digestibility of the sixth crop of alfalfa silage (continued from Table 6)

[0042] Although no fertilization was applied after the fifth alfalfa harvest, the effects of topdressing with organic fertilizer and biopesticides persisted into the sixth harvest. In vitro neutral detergent fiber (NDF) digestibility data showed differences in NDF digestibility between alfalfa silage obtained from the second harvest after fertilization and pesticide application. The digestibility of the chemical fertilizer treatment with chemical pesticides was lower than that of alfalfa silage obtained from the combined application of chemical pesticides and organic fertilizers. The treatment with 1125 g / ha of biopesticides and 3000 kg / ha of organic fertilizers achieved higher digestibility compared to other treatment groups. Compared to other additives and no additives, LP25 significantly improved NDF digestibility, reaching over 64%, representing a 16.11% increase compared to no topdressing, no pesticides, and no additives.

[0043] Table 8. Effects of different treatments on the in vitro digestibility of the sixth crop of alfalfa silage (continued from Table 7)

[0044] Although no fertilization was applied after the fifth alfalfa harvest, the effects of topdressing with organic fertilizer and biopesticides persisted into the sixth harvest. In vitro acid detergent fiber digestibility data showed differences in acid detergent fiber digestibility between alfalfa silage obtained from the second harvest after fertilization and pesticide application. The digestibility of alfalfa silage treated with chemical fertilizers and chemical pesticides was lower than that of alfalfa silage obtained from the combined application of chemical pesticides and organic fertilizers. The digestibility of alfalfa silage treated with 1125 g / ha of biopesticides and 3000 kg / ha of organic fertilizers was significantly higher than other treatment groups. Compared with other additives and no additives, LP25 significantly improved neutral detergent fiber digestibility, reaching over 48%, and compared with no topdressing, no pesticides, and no additives, the improvement in digestibility reached 19.98%.

[0045] In conclusion, the alfalfa silage prepared by adding Lactobacillus paracasei (LP25) to the base of topdressing with organic fertilizer (3000 kg / ha) and biological pesticide (1125 g / ha) yields the best quality.

[0046] This invention discovered the optimal combination of topdressing with organic fertilizer (3000 kg / ha) and biological pesticide (1125 g / ha) from different topdressing measures and pesticide application combinations; it also understood the effects of topdressing measures, pesticide application, and additives on the quality of alfalfa silage, and found that using Lactobacillus plantarum LP25 (i.e. Lactobacillus plantarum Wuzhe25 (CGMCC No. 33705) can significantly improve the fermentation quality and nutritional quality of alfalfa silage.

[0047] By combining organic fertilizers, biopesticides, and additives, dry matter digestibility can be increased by more than 14%, NDF digestibility by more than 15%, and ADF digestibility by more than 19%. The improvement in ADF digestibility is particularly significant.

[0048] Depending on the actual situation, the amount added can be adjusted to achieve faster degradation or a higher degradation rate.

[0049] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A method for improving the digestibility of alfalfa silage in saline-alkali land, characterized in that, The method includes: (1) Topdressing with organic fertilizer and biological pesticides in alfalfa production; (2) Chop the harvested alfalfa raw material, control the moisture content, and inoculate with Lactobacillus plantarum (Lactobacillus plantarum). Lactobacillus plantarum ) Silage is prepared by using microbial agents to obtain alfalfa silage with improved digestibility; The biological pesticide mentioned in step (1) is a Beauveria bassiana biological insecticide wettable powder, the active ingredient of which is Beauveria bassiana (Beauveria bassiana). Beauveria bassiana ); Preferably, the alfalfa is alfalfa grown in saline-alkali land.

2. The method according to claim 1, characterized in that, The organic fertilizer in step (1) has the following composition: organic matter ≥ 30%, N+P2O5+K2O ≥ 4%, wherein the mass ratio of N:P2O5:K2O is 1:1:2; where % is the mass percentage content; the pH of the organic fertilizer is ≤ 8.5; Preferably, the organic matter content is ≥50%.

3. The method according to claim 2, characterized in that, The application rate of organic fertilizer is 3000 kg / ha.

4. The method according to claim 1, characterized in that, Step (1) The bacterial content of Beauveria bassiana in the biological pesticide is 15 billion spores / g.

5. The method according to claim 4, characterized in that, The application rate of biological pesticides is 750-1500 g / ha, preferably 1125 g / ha.

6. The method according to claim 1, characterized in that, Step (1) The application method for organic fertilizer and biological pesticides is as follows: Organic fertilizer is mainly applied starting from the second crop or subsequent crops each year, with additional fertilization applied after every two harvests; and / or, The application of biological pesticides is mainly based on the local pest occurrence patterns, and is carried out during the larval stage or when harmful insects are found.

7. The method according to any one of claims 1-7, characterized in that, Step (2) includes: chopping the harvested alfalfa raw material, controlling the moisture content to 50-70%, inoculating it with Lactobacillus paracasei and / or Lactobacillus plantarum inoculum, and ensiling it at an average temperature of 0-40℃ for 30-360 days to obtain alfalfa silage. Preferably, the Lactobacillus plantarum has the preservation number CGMCC No. 33705.

8. The method according to claim 7, characterized in that, The bacterial content of the bacterial agent is 10. 5 -10 6 cfu / g.

9. Alfalfa silage prepared by the method according to any one of claims 1-8.

10. The application of the alfalfa silage according to claim 9 in animal husbandry; The animals mentioned include cattle and sheep.

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