Methane inhibitor based on plant functional components and use thereof

Abstract

This invention discloses a methane inhibitor based on plant functional components and its application. The inhibitor, by mass fraction (100%), comprises the following raw materials: 1.5-6% p-hydroxybenzoic acid, 0.4-2.4% astragaloside, 5-40% polyethylene glycol 4000, and 81.6-99.999% bentonite. The methane inhibitor provided by this invention uses p-hydroxybenzoic acid and astragaloside as active ingredients. Through their synergistic effect, it effectively inhibits the activity of methanogenic bacteria and the methane production pathway in the rumen of ruminants, while simultaneously improving the rumen fermentation pattern. The above-mentioned methane inhibitor uses natural plant-derived active ingredients, is safe, non-toxic, environmentally friendly, and has a simple preparation process suitable for large-scale production.

Description

Technical Field

[0001] This invention belongs to the field of animal husbandry and environmental protection technology, specifically relating to a methane inhibitor based on plant functional components and its application. Background Technology

[0002] Methane emissions are a major contributor to global warming and climate change. Ruminant microbial fermentation produces large amounts of methane. Global livestock methane emissions increased from 68.0479 million tons in 1961 to 103.5291 million tons in 2019, accounting for 97.5% of total livestock methane emissions and approximately 20% of global methane emissions. Methane's greenhouse effect is dozens of times greater than that of carbon dioxide, and methane emissions also mean a loss of feed energy. Reducing methane emissions from ruminants and improving dietary energy utilization efficiency have attracted widespread attention.

[0003] Currently, measures to reduce rumen methane emissions in ruminants mainly include: adjusting feed nutrient composition, protozoan treatment, adding electron acceptors (such as fumaric acid), adding halogenated compounds and derivatives, and adding ion carriers (such as monensin). However, these methods have problems such as short duration of action, microbial adaptation and tolerance, strong host specificity, reduced fiber digestibility, and toxicity, which greatly limit their application in actual production.

[0004] In recent years, plant secondary metabolites have received widespread attention in the field of methane emission reduction in ruminants due to their advantages such as naturalness, safety, and high efficiency. p-Hydroxybenzoic acid (p-Hydroxybenzoic acid) is a naturally occurring phenolic acid found in many plants, possessing multiple functions including antibacterial and antiseptic properties, influence on microbial flora, and potential pharmacological activities. Studies have found that p-Hydroxybenzoic acid can reduce NH3-N concentration and increase gas production. In the applicant's previous research, p-Hydroxybenzoic acid was found to have the potential to inhibit methane production during rumen fermentation. Kaempferol-3-O-glucoside, a natural flavonoid compound, exhibits various potential pharmacological activities in anti-inflammatory, antioxidant, and neuroprotective effects. It also has inhibitory effects on various microorganisms, and may also inhibit methane-producing bacteria, thereby suppressing methane production. However, there are no reports in the existing technology of combining p-Hydroxybenzoic acid and kaempferol-3-O-glucoside in methane inhibitors, and whether the two have a synergistic effect in inhibiting methane production is also undisclosed.

[0005] In view of this, the present invention is proposed. Summary of the Invention

[0006] To address the aforementioned technical problems, the present invention aims to provide a methane inhibitor based on plant functional components and its application. Using p-hydroxybenzoic acid and astragaloside as active ingredients, the inhibitor effectively inhibits the activity of methanogenic bacteria and the methane generation pathway in the rumen of ruminants through their synergistic effect, while improving the rumen fermentation pattern.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides a methane inhibitor based on plant functional components, comprising, by weight (100%), the following raw materials: 1.5-6% p-hydroxybenzoic acid, 0.4-2.4% astragaloside, 5-40% polyethylene glycol 4000, and 81.6-99.999% bentonite. In the above composition, polyethylene glycol 4000 acts as an encapsulating and diluting dispersant, and bentonite acts as a carrier.

[0008] As a preferred embodiment of the present invention, the methane inhibitor is composed of the following raw materials, with a total mass fraction of 100%: 3-5% p-hydroxybenzoic acid, 0.8-2% astragaloside, 6.5-8.5% polyethylene glycol 4000, and 84.5-89.7% bentonite.

[0009] As a further preferred embodiment of the technical solution of the present invention, the methane inhibitor is composed of the following raw materials, with a total mass fraction of 100%: 3.5~4.5% p-hydroxybenzoic acid, 0.8~2% astragaloside, 7~8% polyethylene glycol 4000, and 85.5~88.7% bentonite.

[0010] As the optimal technical solution of the present invention, the methane inhibitor is, by way of example, composed of the following raw materials: 3% p-hydroxybenzoic acid, 1.6% astragaloside, 5.4% polyethylene glycol 4000, and 90% bentonite.

[0011] Secondly, the present invention provides a method for preparing the above-mentioned methane inhibitor, comprising the following steps: weighing p-hydroxybenzoic acid, astragaloside and polyethylene glycol 4000 in proportion, crushing and sieving polyethylene glycol and mixing it with p-hydroxybenzoic acid and astragaloside and stirring evenly, and then mixing it with bentonite evenly.

[0012] As a preferred embodiment of the technical solution of the present invention, polyethylene glycol 4000 is sieved through a mesh size of 100.

[0013] Thirdly, the present invention also provides the application of the above-mentioned methane inhibitor in the preparation of feed additives or rumen regulators that reduce rumen methane emissions in ruminants.

[0014] As a preferred embodiment of the present invention, when applying it, the aforementioned methane inhibitor can be directly added to the feed at a dosage of 0.5~4 g / kg, and then mixed evenly.

[0015] Fourthly, the present invention also provides a ruminant feed comprising the aforementioned methane inhibitor.

[0016] As a preferred embodiment of the present invention, the methane inhibitor content in the ruminant feed is 0.05-0.5% by mass.

[0017] Compared with the prior art, the present invention has the following beneficial effects: (1) The present invention provides a methane inhibitor based on plant functional components. For the first time, p-hydroxybenzoic acid and astragaloside are combined and applied to methane inhibitors. The two inhibit the activity of rumen methanogens through different mechanisms, exerting a significant synergistic effect. The methane inhibition rate is significantly better than that of a single component.

[0018] (2) The methane inhibitor based on plant functional components provided by the present invention uses natural plant-derived active ingredients, which are safe, non-toxic, green and environmentally friendly, avoiding the toxic side effects and residue problems that may be caused by chemically synthesized inhibitors, and meeting the requirements of green animal husbandry development.

[0019] (3) The present invention provides a methane inhibitor based on plant functional components, which has little impact on normal rumen fermentation and does not affect the production of volatile fatty acids (except acetic acid). While inhibiting methane production, it does not affect feed digestibility and animal production performance.

[0020] (4) The methane inhibitor provided by the present invention has a simple preparation process and low cost, and is suitable for large-scale production and application. Attached Figure Description

[0021] Figure 1 The effect of different treatment groups on the total gas production of in vitro rumen fermentation over 48 hours; Figure 2 The effect of different treatment groups on methane production during 48 hours of in vitro rumen fermentation. Detailed Implementation

[0022] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0023] In this invention, p-hydroxybenzoic acid with a purity ≥98% and astragaloside with a purity ≥98% are both commercially available.

[0024] Polyethylene glycol 4000 was purchased from Guangzhou Meishi Chemical Co., Ltd.

[0025] The bentonite was purchased from Lingshou County Ruifeng Mineral Products Co., Ltd.

[0026] Example 1 A methane inhibitor based on plant functional components is prepared as follows: p-hydroxybenzoic acid, astragaloside, and polyethylene glycol 4000 pulverized through a 100-mesh sieve are weighed in a weight ratio of 35:8:57. The above three substances are mixed, and then the mixture is mixed with bentonite in a weight ratio of 100:920 and stirred evenly to obtain the methane inhibitor.

[0027] Example 2 A methane inhibitor based on plant functional components is prepared as follows: p-hydroxybenzoic acid, astragaloside, and polyethylene glycol 4000 pulverized through a 100-mesh sieve are weighed in a weight ratio of 30:12:58. The three substances are mixed, and then the mixture is mixed with bentonite in a weight ratio of 100:900 and stirred evenly to obtain the methane inhibitor.

[0028] Example 3 A methane inhibitor based on plant functional components is prepared as follows: p-hydroxybenzoic acid, astragaloside, and polyethylene glycol 4000 pulverized through a 100-mesh sieve are weighed in a weight ratio of 30:16:54. The three substances are mixed, and then the mixture is mixed with bentonite in a weight ratio of 100:900 and stirred evenly to obtain the methane inhibitor.

[0029] Example 4 Take the methane inhibitor prepared in Example 3 and mix it evenly into a total mixed ration (the ration consists of the following components by weight: 340 parts corn, 204 parts wheat bran, 75 parts soybean meal, 350 parts corn stalks, 6 parts calcium carbonate, 5 parts salt, and 20 parts premix) at a ratio of 1g per kilogram of dry feed (the concentrate-to-roughage ratio is 65:35) to obtain ruminant feed.

[0030] Example 5 Using artificial rumen in vitro fermentation technology, under the condition of a concentrate-to-roughage ratio of 65:35, a blank control group, a p-hydroxybenzoic acid group (0.0015%), a rutin group (0.0008%), and the composition group of the present invention (p-hydroxybenzoic acid 0.0015%, rutin 0.0008%, polyethylene glycol 4000 0.0035%, bentonite 99.9942%) were set up, and the cumulative methane yield of each group was measured after 48 hours.

[0031] Experimental method: Rumen fluid was collected, filtered through four layers of gauze, and mixed with artificial saliva at a ratio of 1:4. The mixture was then dispensed into anaerobic fermentation bottles, and appropriate additives and substrates were added. The mixture was incubated at 39°C for 48 hours, and the methane content was determined by collecting the gas.

[0032] Experimental results: The results are as follows Figure 1 and Figure 2 As shown, there was no significant difference in cumulative methane production among the four groups (P>0.05). Compared with the blank control group, the methane production in the p-hydroxybenzoic acid group decreased by approximately 9.19%, the methane production in the astragaloside group decreased by approximately 12.42%, while the methane production in the composition group of the present invention decreased by approximately 26.6%. The methane inhibition effect of the composition was significantly better than that of each individual component (P<0.05), indicating that the two have a significant synergistic effect in inhibiting methane formation.

[0033] Example 6 Based on the experiment in Example 5, fermentation parameters for each group were measured, and the specific results are shown in Table 1. The results showed that after adding the methane inhibitor of this invention, the NH3-N concentration was the lowest and significantly lower than that of the control group (P<0.05); it had no significant effect on the total volatile fatty acid concentration, or the molar percentages of acetic acid and butyric acid (P>0.05), but the propionic acid ratio significantly increased relative to the control group and p-hydroxybenzoic acid (P<0.05), while the ethyl-to-propionic acid ratio significantly decreased (P<0.05). This indicates that the methane inhibitor of this invention, while inhibiting methane production, can transform rumen fermentation from acetic acid-type fermentation to propionic acid-type fermentation, thus promoting the growth of ruminant livestock for meat production.

[0034] Table 1. Effects of methane inhibitors on rumen fermentation parameters (DMD, pH, NH3-N, VFA) Example 7 Twenty healthy sheep of similar weight were randomly divided into a control group and an experimental group, with 10 sheep in each group. The control group was fed a basal diet, while the experimental group was fed a basal diet supplemented with the methane inhibitor prepared in Example 3 of this invention (at a dosage of 1 g per kilogram of dry matter). The pre-feeding period was 7 days, and the trial period was 28 days. Methane emissions were measured using the respiratory metabolism chamber method. The test results are shown in Table 2.

[0035] Results: The daily methane emissions of sheep in the experimental group were 23.9% lower than those in the control group (P<0.05), while there was no significant difference in dry matter intake (P>0.05), and feed conversion ratio was improved. The sheep maintained good condition throughout the experiment, and no adverse reactions were observed.

[0036] Table 2. Effects of methane inhibitors on feed intake and methane emissions in sheep. The applicant declares that the present invention is illustrated by the above embodiments, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent substitutions of individual raw materials in the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A methane inhibitor based on plant functional components, characterized in that, It is composed of the following raw materials in a total mass fraction of 100%: 1.5~6% p-hydroxybenzoic acid, 0.4~2.4% astragaloside, 5~40% polyethylene glycol 4000, and 81.6~99.999% bentonite.

2. The methane inhibitor based on plant functional components according to claim 1, characterized in that, The methane inhibitor, in 100% by mass fraction, is composed of the following raw materials: 3-5% p-hydroxybenzoic acid, 0.8-2% astragaloside, 6.5-8.5% polyethylene glycol 4000, and 84.5-89.7% bentonite.

3. A methane inhibitor based on plant functional components according to claim 1, characterized in that, The methane inhibitor, in 100% by mass fraction, is composed of the following raw materials: 3.5-4.5% p-hydroxybenzoic acid, 0.8-2% astragaloside, 7-8% polyethylene glycol 4000, and 85.5-88.7% bentonite.

4. A methane inhibitor based on plant functional components according to claim 1, characterized in that, The methane inhibitor is composed of the following raw materials, with a total mass fraction of 100%: 3% p-hydroxybenzoic acid, 1.6% astragaloside, 5.4% polyethylene glycol 4000, and 90% bentonite.

5. A method for preparing the methane inhibitor according to any one of claims 1 to 4, characterized in that, The process includes the following steps: Weigh p-hydroxybenzoic acid, astragaloside, and polyethylene glycol 4000 according to the specified proportions. After crushing and sieving the polyethylene glycol, mix it with p-hydroxybenzoic acid and astragaloside and stir until homogeneous. Then, mix it with bentonite until homogeneous.

6. The preparation method according to claim 5, characterized in that, Polyethylene glycol 4000 has a sieve mesh size of 100.

7. The use of the methane inhibitor according to any one of claims 1 to 4 in the preparation of feed additives or rumen regulators that reduce rumen methane emissions in ruminants.

8. The application according to claim 7, characterized in that, The methane inhibitor described in any one of claims 1 to 4 is added to the feed at a dosage of 0.5 to 4 g / kg and mixed evenly.

9. A ruminant feed, characterized in that, It includes the methane inhibitor according to any one of claims 1 to 4.

10. The ruminant feed according to claim 9, characterized in that, The mass content of the methane inhibitor is 0.05~0.5%.

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