Application of hydrogen-rich alkaline electrolysis water in promoting rumen fermentation of beef cattle drinking water

Abstract

The application discloses application of alkaline hydrogen-rich electrolytic water in promoting rumen fermentation of beef cattle drinking water and belongs to the technical field of poultry and livestock drinking water additives. The alkaline hydrogen-rich electrolytic water is used as the drinking water for promoting rumen fermentation of beef cattle, so that the emission of methane in the in-vitro rumen fermentation of beef cattle is effectively reduced, the pH value of the in-vitro rumen fermentation liquid is improved, the crude protein content of the in-vitro rumen fermentation microorganism of beef cattle is increased, and the rumen microbial flora is improved, and finally the rumen fermentation efficiency is improved. The beef cattle can drink the alkaline hydrogen-rich electrolytic water used in the application, which can not only reduce the emission of methane in the in-vitro rumen fermentation of beef cattle, effectively improve the rumen fermentation efficiency, but also has no toxic side effects and is environment-friendly, is a very good antibiotic substitute, and avoids problems such as drug residues existing in the prior art antibiotics.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of livestock drinking water additives, and particularly relates to application of alkaline hydrogen-rich electrolytic water in promoting rumen fermentation of beef cattle. BACKGROUND

[0002] Water consumption and water quality are important guarantees for maintaining the metabolism and immunity of beef cattle and are closely related to the growth performance of beef cattle. However, the underground water of livestock and poultry farms is often contaminated by livestock and poultry manure and contains a large number of pathogenic microorganisms such as leptospira, salmonella, tularemia, anthrax bacillus, enterovirus and adenovirus. If livestock and poultry drink the underground water containing these pathogenic microorganisms, infectious diseases may break out. Studies have shown that high concentrations of total dissolved solids (CFU>1000mg / L) and total bacterial count (CFU>100mg / mL) in drinking water can reduce the production performance and disease resistance of livestock and poultry. Therefore, controlling the water quality of beef cattle is one of the important ways to promote the healthy and efficient production of beef cattle.

[0003] Electrolytic water refers to acid, neutral and alkaline functional water produced by electrolysis of dilute sodium chloride, potassium chloride, magnesium chloride and dilute hydrochloric acid solution alone or in mixture in an electrolysis chamber. In recent years, electrolytic water has been widely used in the food industry due to its efficient antibacterial activity, low operating cost and safety without pollution and residue. As a research focus of electrolytic water in recent years, alkaline hydrogen-rich electrolytic water has important significance for the relief of chronic heat stress of livestock and poultry and the enhancement of the antioxidant capacity of the body. Specifically, H2 in alkaline hydrogen-rich electrolytic water can rapidly diffuse through the lipid membrane to various subcellular structures and help maintain the stability of the respiratory chain in mitochondria, increase the concentration of coenzyme Q9 in the mitochondrial electron transport chain and promote the oxidative phosphorylation of cell mitochondria to produce ATP. On the other hand, H2 can regulate gene expression through histone modification and induce mitochondrial unfolded protein response, thereby relieving the mitochondrial oxidative stress caused by the unfolded protein response. However, there is no report on the application of alkaline hydrogen-rich electrolytic water as drinking water to regulate rumen fermentation of beef cattle, and it is unknown whether the rumen fermentation performance of beef cattle can be improved. SUMMARY

[0004] The application aims to provide application of alkaline hydrogen-rich electrolytic water in promoting rumen fermentation of beef cattle. The use of alkaline hydrogen-rich electrolytic water as drinking water for promoting rumen fermentation of beef cattle can effectively reduce the emission of methane in beef cattle rumen in vitro fermentation, increase the pH value of rumen in vitro fermentation liquid, increase the crude protein content of rumen in vitro fermentation microorganisms and improve the rumen microbial flora, and ultimately improve the rumen fermentation efficiency.

[0005] To achieve the above-mentioned purpose, the application provides the following technical scheme.

[0006] The application of an alkaline hydrogen-rich electrolytic water in promoting rumen fermentation of beef cattle, wherein the concentration of H2 in the alkaline hydrogen-rich electrolytic water is 400±50ppb.

[0007] Preferably, the alkaline hydrogen-rich electrolytic water is obtained by electrolysis of distilled water through a negative ion water machine.

[0008] The beneficial technical effects of the present application are as follows:

[0009] The present application uses an alkaline hydrogen-rich electrolytic water that is drinkable for beef cattle, which not only reduces the emission of methane in the in vitro fermentation of beef cattle rumen, effectively improves the rumen fermentation efficiency, but also has no toxic side effects and is environmentally friendly, and is a very good antibiotic substitute, avoiding the problems of drug residues and the like existing in the existing antibiotics. BRIEF DESCRIPTION OF DRAWINGS

[0010] Figure 1 The effects of preparing rumen fermentation liquid with different concentrations of alkaline hydrogen-rich electrolytic water on total gas production and methane production, wherein A is the total gas production of each group after 12h of fermentation, B is the methane production of each group after 12h of fermentation, C is the total gas production of each group after 48h of fermentation, and D is the methane production of each group after 48h of fermentation.

[0011] Figure 2 The PCoA analysis results of the effects of 400ppb alkaline hydrogen-rich electrolytic water on the beta diversity of the fermentation liquid, wherein A is the PCoA analysis result after 12h of fermentation, and B is the PCoA analysis result after 48h of fermentation.

[0012] Figure 3 The bar charts of the species composition at the door level of the DW group and the HRW group, wherein A is the bar chart of the species composition at the door level after 12h of fermentation, and B is the bar chart of the species composition at the door level after 48h of fermentation.

[0013] Figure 4 The difference analysis results of the dominant bacteria at the door level after 12h of fermentation.

[0014] Figure 5 The difference analysis results of the dominant bacteria at the door level after 48h of fermentation.

[0015] Figure 6 The bar charts of the species composition at the genus level of the DW group and the HRW group, wherein A is the bar chart of the species composition at the genus level after 12h of fermentation, and B is the bar chart of the species composition at the genus level after 48h of fermentation.

[0016] Figure 7 The difference analysis results of the dominant bacteria at the genus level after 12h of fermentation.

[0017] Figure 8 The difference analysis results of the dominant bacteria at the genus level after 48h of fermentation. DETAILED DESCRIPTION

[0018] The following detailed description of various example embodiments of the application should not be considered to be limiting of the application, but rather a description of certain example aspects, features and embodiments of the application. It is understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the application.

[0019] Further, for numerical ranges expressed in the disclosure, it is understood that every intervening value, to the upper and lower limits, is also specifically disclosed. In any stated range the intervening values include each and every sub-range therein. These sub-ranges include unitary values or discrete values. The upper and lower limits of these intervening values are also disclosed. The upper and lower limits of any of the ranges specifically and explicitly disclosed are also individually and explicitly disclosed.

[0020] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, the preferred methods and materials are described.

[0021] As used herein, the terms "comprise", "comprising", "include", "including", "have", "having" and the like are open-ended terms that are intended to permit but not limit the amount or type of ingredients or steps to be included.

[0022] Example 1

[0023] Preparation of 800 ppb alkaline hydrogen-rich electrolyzed water:

[0024] 2 L of distilled water was added into the negative ion water machine (V8 type, Zhongshan Li Fu Ren Appliance Co., Ltd.) using a measuring cylinder, and alkaline hydrogen-rich electrolyzed water was prepared by electrifying for 0.5 h. The obtained alkaline hydrogen-rich electrolyzed water had a pH of 8.69, an ORP of -554 mV, and a hydrogen concentration of 0.81 mg / L.

[0025] Example 2

[0026] Preparation of 400 ppb alkaline hydrogen-rich electrolyzed water:

[0027] 1 L of the 800 ppb alkaline hydrogen-rich electrolyzed water prepared in Example 1 was taken and 1 L of distilled water was added to obtain 400 ppb alkaline hydrogen-rich electrolyzed water. The obtained alkaline hydrogen-rich electrolyzed water had a pH of 7.64, an ORP of -72 mV, and a hydrogen concentration of 0.44 mg / L.

[0028] Example 3

[0029] Preparation of 200 ppb alkaline hydrogen-rich electrolyzed water:

[0030] Take 400 ppb of the alkaline hydrogen-rich electrolytic water prepared in Example 2, add 1 L of distilled water, to obtain 200 ppb of the alkaline hydrogen-rich electrolytic water, the obtained alkaline hydrogen-rich electrolytic water has a pH of 7.52, an ORP of -14 mv, and a hydrogen concentration of 0.22 mg / L.

[0031] Test Example

[0032] The alkaline hydrogen-rich electrolytic water prepared in Examples 1-3 was used to prepare rumen fermentation liquid.

[0033] First, artificial saliva was prepared, and the preparation steps were as follows:

[0034] Take 520.2 mL of the alkaline hydrogen-rich electrolytic water prepared in each example, and sequentially add 208.1 mL of buffer solution (4.0 g of NH4HCO3+ 35 g of NaHCO3, dissolved in distilled water, and diluted to 1000 mL), 208.1 mL of macroelement solution (9.45 g of Na2HPO4·12H2O+ 6.2 g of anhydrous KH2PO4+ 0.6 g of MgSO4·7H2O, dissolved in distilled water, and diluted to 1000 mL), 0.1 mL of trace element solution (13.2 g of CaCl2·2H2O+ 10.0 g of MnCl2·4H2O+ 1.0 g of CoCl2·6H2O+ 8.0 g of FeCl3·6H2O, dissolved in distilled water, and diluted to 1000 mL), and 62.4 mL of reducing solution (4.0 mL of 1 mol / L NaOH+ 625 mg of Na2S·9H2O+ 625 mg of cysteine hydrochloride+ 95 mL of distilled water), and pass carbon dioxide until the solution changes from light blue to colorless.

[0035] Preparation of rumen fermentation liquid:

[0036] The artificial saliva and fresh rumen liquid collected through a rumen fistula were mixed in a volume ratio of 2:1 to prepare the rumen fermentation liquid.

[0037] Feed dry sample:

[0038] The weight percentage of raw materials: wheat straw 50.71%, corn 20.00%, wheat bran 3.49%, soybean meal 19.80%, calcium bicarbonate 0.50%, calcium hydrogen phosphate 0.50%, premix 4.00%, and salt 0.50%; wherein, the premix contains vitamin A 100000 IU, vitamin D3 2700 IU, vitamin E 800 IU, 900 mg of Fe, 1100 mg of Mn, 1200 mg of Zn, 250 mg of Cu, 120 g of Ca, and 35 g of P per kilogram; and the powder was crushed through a 1 mm sieve to obtain the feed dry sample.

[0039] Investigation of the effect of rumen fermentation liquid prepared with the basic hydrogen-rich electrolytic water of examples 1-3 on rumen fermentation characteristics

[0040] (1) Test design

[0041] A single-factor test design was used, and a control group in which distilled water was used instead of basic hydrogen-rich electrolytic water to prepare the rumen fermentation liquid was set up, denoted as the DW group; a HRW 200ppb group in which the rumen fermentation liquid was prepared with 200 ppb basic hydrogen-rich electrolytic water of example 3; a HRW 400ppb group in which the rumen fermentation liquid was prepared with 400 ppb basic hydrogen-rich electrolytic water of example 2; and a HRW 800ppb group in which the rumen fermentation liquid was prepared with 800 ppb basic hydrogen-rich electrolytic water of example 1.

[0042] 60 mL of the rumen fermentation liquid of each group was taken, 0.500 g of a feed air-dried sample was added, and the mixture was placed in a constant-temperature shaker to simulate rumen fermentation. There were 10 replicates in each group, and fermentation was stopped at 12 h for 5 replicates and at 48 h for the other 5 replicates.

[0043] (2) Sample collection

[0044] After 12 h and 48 h of culture, the fermentation was terminated in an ice bath (5 replicates at each time point in each group), the gas production was determined, the fermentation liquid was filtered, the pH value of the fermentation liquid was determined, the fermentation liquid was collected and stored at -20℃, and the concentrations of ammonia nitrogen (NH3-N), microbial crude protein (MCP), and volatile fatty acid (VFA) were determined. The filter residue was transferred into a nylon bag without damage, and was dried in an oven at 65℃ to determine the dry matter content.

[0045] (3) Index determination

[0046] ① Net gas production and gas production parameters: at 3, 6, 9, 12, 18, 24, 27, 30, 36, and 48 h of culture, the culture tubes were taken out, the scale value (mL) at which the piston was located was quickly read and recorded, and the net gas production in each time period was calculated, net gas production (mL) = gas production (mL) in a certain time period - blank gas production (mL) in the same time period.

[0047] ② Dry matter: the dry matter at 12 h and 48 h was dried at 65℃ for 48 h, and was weighed after rehydration for 24 h.

[0048] ③ Determination of NH3-N concentration: a 752 ultraviolet-visible spectrophotometer was used for the determination.

[0049] (iv) VFA concentration: After the fermentation broth filtered by four layers of gauze was centrifuged at 10,000 r / min for 15 min, 1 mL of supernatant was mixed with 200 μL of 25% metaphosphoric acid, filtered through a membrane, and then detected on a machine. Gas chromatography (Shimadzu GC-2014, Japan) was used to determine the content of acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid and valeric acid, and the sum of the six volatile fatty acids was the total volatile fatty acid, and the sum of isobutyric acid and isovaleric acid was defined as branched-chain volatile fatty acid. The peak area method was used to identify and convert the content of various volatile fatty acids based on the relative retention time. The standard curve was prepared under the same conditions using the same method.

[0050] (v) MCP: The content of MCP in the fermentation broth was determined by Coomassie brilliant blue method and UV-visible spectrophotometer.

[0051] (vi) Methane production (CH4): The methane production was estimated by the formula described by Moss et al. (2000).

[0052] CH4(mmol / L) = 0.45 x C2-0.275 x C3+0.40 x C4

[0053] In the formula: C2 = concentration of acetic acid (mmol / L); C3 = concentration of propionic acid (mmol / L); C4 = concentration of butyric acid (mmol / L).

[0054] (vii) 16S rRNA high-throughput sequencing

[0055] Due to HRW 400ppb groups had a significant effect on rumen fermentation characteristics in vitro, and HRW 200ppb groups and HRW 800ppb groups did not have a significant effect on the electrolytic water at a concentration of 400 ppb, so the fermentation broth of the DW group (distilled water control group) and the HRW 400ppb group (referred to as HRW group in 16sRNA related data) was subjected to 16S rRNA high-throughput sequencing, and the test method was as follows:

[0056] DNA extraction was used to determine the quantity and quality of DNA using NanoDrop 2000 (NanoDrop Technologic, Wilmington, DE, USA); for bacteria, the present application used universal primers (338F and 806R) to amplify the V3-V4 region of the 16S rRNA gene for PCR amplification, which was performed using a 20 mL TranStart Fastpfu DNA polymerase (TransGen Biontech, Beijing, China) reaction system, including 4 microliters of 5x FastPfu buffer, 2 microliters of 2.5 mmol / L dNTPs, 0.8 microliters of 5 mmol / L upstream primer, 0.8 microliters of 5 mmol / L downstream primer, 0.4 microliters of DNA polymerase, and 10 ng of template DNA. The amplification program was as follows: first pre-denaturation at 95°C for 5 minutes, then 27 cycles of 95°C for 30 seconds, annealing at 55°C for 30 seconds, extension at 72°C for 45 seconds, and finally extension at 72°C for 10 minutes, and storage at 4°C.

[0057] After PCR amplification, all amplicon libraries were sequenced using the Illumina Miseq PE 300 platform. The raw sequences were processed using Trimmomatic software, and bases with a quality score of more than 20 were retained. FLASH software was used to assemble the end reads. Then, UPARSE (Version 7.0.1) was used for OUT clustering at a similarity threshold of 97%, and RDP classifier was used for species classification annotation of each sequence, with a bacterial database of SILVA (Version 11.9) and a set alignment threshold of 70%. A dilution curve was constructed to ensure sufficient sequencing depth, and Chao, Shannon, and Simpson indices were used to determine bacterial richness and diversity. QIME was used for beta diversity calculation. Principal coordinate analysis (PCoA) was based on the Bray-Curtis similarity matrix, and AMOVA variance analysis was performed by MOTHUR (Version 1.29) to evaluate the significant differences between each sample.

[0058] (4) Data processing and statistical analysis

[0059] SPSS19.0 was used for one-way ANOVA analysis of the three treatment groups, and the data were statistically analyzed. The results were expressed as means, and P<0.05 was considered statistically significant.

[0060] Test results

[0061] (1) Effects of different concentrations of alkaline hydrogen-rich electrolyzed water on total gas production and methane production

[0062] The test results are shown below. Figure 1 Where A represents the total gas production of each group after 12 hours of fermentation, B represents the methane production of each group after 12 hours of fermentation, C represents the total gas production of each group after 48 hours of fermentation, and D represents the methane production of each group after 48 hours of fermentation. Figure 1 It can be seen that with the increase of HRW concentration, the total gas production and CH4 production at 12h and 48h in each group showed a trend of first decreasing and then increasing. The total gas production at 12h in the 800ppb group was significantly higher than that in other groups. Figure 1 In the middle group (A, P<0.01), the total gas production over 48 hours was significantly higher than that of the 200 ppb group (). Figure 1 The concentration of C (P<0.01) was significantly higher than that of the 400 ppb group. Figure 1 (P<0.05) There was no significant difference in total gas production among the control group, the 200ppb group, and the 400ppb group at 12h and 48h of fermentation (P>0.05); however, the CH4 production in the 200ppb and 400ppb groups was significantly lower than that in the control group at 12h. Figure 1 In the 200 ppb group (P < 0.05), the CH4 yield at 48 h was significantly lower than that of other groups. Figure 1 (D, P<0.01), the CH4 yield at 48 h in the 400 ppb group was significantly lower than that in the 800 ppb group ( Figure 1 (2) Effects of different concentrations of alkaline hydrogen-rich electrolyzed water on the in vitro fermentation characteristics of rumen (P<0.05).

[0063] The results of the determination of various fermentation characteristic indicators are shown in Table 1. As can be seen from Table 1, compared with the control group, after 12 h of in vitro fermentation, the pH and NH3-N concentration of the three treatment groups increased significantly (P<0.05), and showed a decreasing trend with the increase of HRW concentration. The NH3-N concentration of the 200 ppb group was significantly higher than that of the 800 ppb group (P<0.05), but there was no significant difference compared with the 400 ppb group (P>0.05). The dry matter degradation rate of the 200 ppb group was significantly lower than that of the other groups (P<0.05). The MCP content of the 400 ppb group was significantly higher than that of the 200 ppb and 800 ppb groups (P<0.05). The MCP content of the 800 ppb group was significantly lower than that of the other groups (P<0.05). After 48 hours of fermentation, the pH and MCP content of the 200 ppb and 400 ppb groups were significantly higher than those of the control group and the 800 ppb group (P<0.05); the NH3-N concentration of the 200 ppb and 800 ppb groups was significantly lower than that of the control group (P<0.05), while there was no significant difference between the 400 ppb group and the control group (P>0.05); different concentrations of HRW had no significant effect on the dry matter degradation rate (P>0.05).

[0064] Table 1 Effect of different concentrations of alkaline hydrogen-rich electrolyzed water on rumen fermentation characteristics in vitro

[0065]

[0066]

[0067] Note: The same row data shoulder mark different letters represent significant difference (P <0.05), the same letter or no letter represents no significant difference (P >0.05); the same below.

[0068] (3) Effect of different concentrations of alkaline hydrogen-rich electrolyzed water on rumen fermentation VFA in vitro

[0069] The determination results of VFA indexes of each group are shown in Table 2. It can be seen from Table 2 that the acetic acid content of the 200 ppb group was significantly lower than that of the other groups (P <0.05), and the propionic acid content was significantly lower than that of the control group (P <0.05), and the propionic acid content of the 800 ppb group was the lowest (P <0.05); the isobutyric acid content of the 200 ppb group was significantly lower than that of the other groups (P <0.05) at 48 h, and the butyric acid content and acetic acid / propionic acid value of the 800 ppb group were significantly higher than those of the other groups (P <0.05); the pentanoic acid and iso-pentanoic acid contents of the control group were significantly higher than those of the 200 ppb group and the 800 ppb group (P <0.05) at 48 h, and there was no significant difference between the control group and the 400 ppb group (P >0.05); the TVFA content of the control group was significantly higher than that of the other groups (P <0.05), and the 200 ppb group was significantly lower than the other groups (P <0.05), and there was no significant difference between the 400 ppb group and the 800 ppb group (P >0.05); the non-gluconic acid / glucose acid value of the 800 ppb group was the highest, and the fermentation efficiency was significantly lower than that of the control group and the 200 ppb group (P <0.05), and the non-gluconic acid / glucose acid value of the 200 ppb group was significantly lower than that of the other groups (P <0.05) at 48 h, and the fermentation efficiency was significantly higher than that of the other groups (P <0.05).

[0070] Table 2 Effect of 3% acidic electrolyzed water and alkaline electrolyzed water on rumen fermentation total volatile fatty acids

[0071]

[0072]

[0073] After comprehensive consideration, the 400 ppb alkaline hydrogen-rich electrolyzed water prepared in Example 2 has the best effect on promoting rumen fermentation of beef cattle, and the effect of the alkaline hydrogen-rich electrolyzed water on the fermentation liquid bacteria flora is further investigated.

[0074] (4) Effect of 400 ppb alkaline hydrogen-rich electrolyzed water on fermentation liquid bacteria flora alpha diversity analysis

[0075] The results of each index are shown in Table 3. As shown in Table 3, after 12h fermentation, the differences in Sobs, Chao and Ace indexes between the two groups were not significant, indicating that the differences in microbial population, microbial diversity and microbial community richness between the two groups were not significant (P>0.05). After 48h fermentation, the difference in Ace index between the two groups was not significant, indicating that the difference in microbial community richness was not significant (P>0.05). After 12h fermentation, compared with the DW group, the Simpson index of the HRW group increased significantly, and the Shannon index decreased significantly (P<0.05), indicating that the microbial community richness and uniformity and community diversity of the HRW group were significantly lower than those of the DW group. After 48h fermentation, compared with the DW group, the Sobs, Shannon and Chao indexes of the HRW group increased significantly, and the Simpson index decreased significantly (P<0.05), indicating that the community diversity, microbial population and microbial community richness and uniformity of the HRW group were lower than those of the DW group. 400ppb

[0076] Table 3400ppb alkaline hydrogen-rich electrolytic water on the effect of fermentation broth microbial α diversity analysis

[0077]

[0078] (5)400ppb alkaline hydrogen-rich electrolytic water on the effect of fermentation broth microbial β diversity analysis

[0079] The PCoA analysis results are shown in Figure 2 , wherein A is the PCoA analysis result of 12h fermentation, and B is the PCoA analysis result of 48h fermentation. As shown in Figure 2 A, the samples of the DW group and the HRW group are relatively discrete, indicating that there is a large difference between different samples in the two groups; R=0.9660 is close to 1, indicating that the difference between the two groups is greater than the difference within the group. P<0.05, indicating that the test is highly reliable. As shown in Figure 2 B, the 5 samples of the DW group are relatively discrete, while the 5 samples of the HRW group are more densely distributed, indicating that the difference between different samples in the HRW group is smaller than that in the DW group; R=0.6600 is close to 1, indicating that the difference between the two groups is greater than the difference within the group. P<0.05, indicating that the test is highly reliable.

[0080] (6)400ppb alkaline hydrogen-rich electrolytic water on the effect of fermentation broth microbial species composition

[0081] ① Species abundance analysis at the phylum level

[0082] The phylum level species composition bar chart of the DW group and the HRW group is shown in Figure 3 , wherein A is the phylum level species composition bar chart of 12h fermentation, and B is the phylum level species composition bar chart of 48h fermentation. As shown in​Figure 3 As shown in the figure, the door level flora in the rumen microorganism is mainly Firmicutes, Bacteroidota, Proteobacteria.

[0083] The present application carries out difference analysis on dominant bacteria at the door level Figure 4 And Figure 5 At the fermentation time of 12h Figure 4 Compared with the DW group, there are 5 doors with significant differences in relative abundance in the HRW group, among which the relative abundance of Firmicutes, Synergistota and Spirochaetota is significantly increased (P<0.05), and the relative abundance of Bacteroidota and Proteobacteria is significantly reduced (P<0.05). The relative abundance of other doors has no significant difference. At the fermentation time of 48h Figure 5 Compared with the DW group, there are 6 doors with significant differences in relative abundance in the HRW group, among which the relative abundance of Fusobacteriota is significantly increased (P<0.05), and the relative abundance of Desulfobacterota, Synergistota, Verrucomicrobiota, Actinobacteriota and Patescibacteria is significantly reduced (P<0.05). The relative abundance of other doors has no significant difference.

[0084] 2. Species abundance analysis at the genus level

[0085] The columnar chart of the species composition at the genus level of the DW group and the HRW group is shown in Figure 6 , wherein A is the columnar chart of the species composition at the genus level of the fermentation of 12h, and B is the columnar chart of the species composition at the genus level of the fermentation of 48h. As shown in Figure 6 , at the genus level of the species composition, the microorganisms in the fermentation broth are mainly composed of Rikenellaceae-RC9-gut-group, Prevotella, Streptococcus, Selenomonas, Succinivibrio, Ruminobacter, Schwartzia, Prevotellaceae-YAB2003-group, Oribacterium and the like.

[0086] The present application carries out difference analysis on dominant bacteria at the genus level Figure 7 And Figure 8 At the fermentation time of 12h Figure 7), compared with the DW group, 7 genera in the HRW group had significant differences in relative abundance, among which Streptococcus, Shigella and Prevotellaceae YAB2003 group had significantly increased relative abundance (P<0.05), and Prevotella, Selenomonas, Succinivibrio and unclassified-f-Succinivibrionaceae had significantly decreased relative abundance (P<0.05). The relative abundance of other genera had no significant difference. At 48 h of fermentation Figure 8 ), compared with the DW group, 3 genera in the HRW group had significant differences in relative abundance, among which Prevotellaceae YAB2003 group and Oribacterium had significantly increased relative abundance (P<0.05), and Roseburia had significantly decreased relative abundance (P<0.05). The relative abundance of other genera had no significant difference.

[0087] In summary Figures 3 to 8 As can be seen by those skilled in the art, at 12 h of fermentation, by adding hydrogen-rich water, the composition of rumen microorganisms changed significantly, the abundance of cellulose-degrading flora such as Firmicutes and Spirochaetes increased, which promoted rumen fermentation by increasing the degradation of cellulose and non-cellulose polysaccharides; Synergistetes may be involved in the reduction of sulfate and nitrate, optimizing the utilization of sulfate and nitrate and participating in the degradation of protein to promote rumen fermentation. The abundance of important flora such as Bacteroidetes and Proteobacteria decreased, which may have an adverse effect on the degradation of polysaccharides and proteins, the production of short-chain fatty acids and the maintenance of rumen microbial homeostasis. The increase in the abundance of lactic acid bacteria such as Streptococcus may improve the utilization of nutrients, while the decrease in the abundance of protein / carbohydrate-degrading bacteria such as Prevotella may reduce the degradation of polysaccharides and proteins, affecting nitrogen cycling. At 48 h of fermentation, by adding hydrogen-rich water, the abundance of microorganisms involved in cellulose and fibrous polysaccharide degradation (Fusobacteria) increased, which improved feed utilization; the abundance of microorganisms involved in the production of excess hydrogen sulfide (Desulfobacteres) decreased, which may be beneficial to maintaining the stability of the rumen environment. At the same time, some protein / carbohydrate-degrading bacteria (Actinobacteria, Oribacterium and Roseburia) changed, which may affect protein degradation and protein metabolism. In addition, the effects of certain microorganisms (Patescibacteria and Prevotellaceae YAB2003 group) on rumen function are not yet clear and need further study. Overall, the addition of HRW changed the composition of rumen microbial flora, increased the relative abundance of Firmicutes, Spirochaetes and Fusobacteria, reduced hydrogen sulfide-producing bacteria (Desulfobacteres), increased the utilization of cellulose and fibrous polysaccharides, improved rumen fermentation efficiency and reduced methane production.

[0088] The above described embodiments are only to illustrate the preferred modes of the present application, and are not intended to limit the scope of the present application. Any modification and improvement made by those skilled in the art to the technical solutions of the present application without departing from the design spirit of the present application shall fall within the protection scope of the present application as defined by the claims.

Claims

1. The use of alkaline hydrogen-rich electrolytic water in the preparation of rumen fermentation drinking water for beef cattle, characterized in that, The application is to promote rumen fermentation of beef cattle by using the alkaline hydrogen-rich electrolytic water to reduce total gas production and methane production during rumen fermentation of beef cattle; the concentration of H2 in the alkaline hydrogen-rich electrolytic water is 400±50ppb.

2. Use of the basic hydrogen-rich electrolyzed water according to claim 1 for preparing rumen fermentation drinking water for beef cattle, characterized in that, The alkaline hydrogen-rich electrolytic water is obtained by electrolysis of distilled water by a negative ion water machine.

Images