CN113854406A - Application of nitrate in improving morphological development of gastrointestinal tract tissue of Hu sheep and improving microorganisms - Google Patents
Application of nitrate in improving morphological development of gastrointestinal tract tissue of Hu sheep and improving microorganisms Download PDFInfo
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- CN113854406A CN113854406A CN202110981618.0A CN202110981618A CN113854406A CN 113854406 A CN113854406 A CN 113854406A CN 202110981618 A CN202110981618 A CN 202110981618A CN 113854406 A CN113854406 A CN 113854406A
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- calcium nitrate
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Images
Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/33—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from molasses
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/174—Vitamins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/22—Compounds of alkali metals
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/24—Compounds of alkaline earth metals, e.g. magnesium
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
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- A23K20/30—Oligoelements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
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Abstract
The invention provides application of nitrate in improving gastrointestinal tract tissue morphological development and microorganism improvement of Hu sheep, 1-3% of nitrate is added into feed to feed Hu sheep, so that the tissue morphological development of the gastrointestinal tract of Hu sheep can be effectively improved, the population of microorganisms can be improved, the weight of the body, head, hoof and internal organs of Hu sheep can be effectively improved, the fiber decomposition capacity of colon can be improved, and the feed can be better absorbed and utilized.
Description
Technical Field
The invention belongs to the technical field, and particularly relates to application of nitrate in improving the morphological development of gastrointestinal tract tissues of Hu sheep and improving microorganisms.
Background
The rumen of ruminants plays a critical role in the digestive process, and is rich in various microorganisms. After the animals eat the feed, the forage enters the rumen, various nutrient substances in the forage are degraded under the action of rumen microorganisms, and degradation products are converted into various nutrient substances and energy required by the animals after being absorbed by the microorganisms. Rumen microorganisms can degrade crude fiber and carbohydrates into Volatile Fatty Acids (VFAs) and release energy. The protein in the feed can be decomposed into amino acid and small peptide, and then the amino acid, the small peptide and other non-protein nitrogen in rumen are utilized to synthesize the bacterial protein with higher quality. Nitrate has been reported to have an effect on ruminal fermentation in ruminants. After ruminants eat a feed containing nitrate, rumen microorganisms firstly reduce the feed into nitrite through decomposition under the anaerobic condition in rumen, and finally generate ammoniacal nitrogen. In the process that nitrate generates ammonia nitrogen through nitrite in rumen, the generation of propionic acid can be inhibited through consuming hydrogen, and the proliferation of fiber degrading bacteria is promoted, so that the fermentation mode of rumen is influenced. But morphological development and microbial changes of gastrointestinal tract tissues of the Hu sheep have not been reported.
Disclosure of Invention
In order to solve the technical problems, the invention provides the application of nitrate in improving the morphological development of gastrointestinal tract tissues of Hu sheep and improving microorganisms.
In order to achieve the purpose, the invention adopts the following technical scheme that:
the nitrate is applied to improving the morphological development of the gastrointestinal tract tissue of the Hu sheep and improving the microorganism.
In the application, the nitrate is preferably used in an amount of 1-3% of the feed.
The application is preferably that the feed comprises 15-18 parts of corn straws, 9-11 parts of soybean hulls, 1-3 parts of sunflower hulls, 45-50 parts of corns, 10-12 parts of soybean meal, 2-4 parts of sesame cakes, 3-6 parts of molasses, 0.2-0.5 part of mountain flour, 0.60-0.8 part of NaCl, 0.5-1.50 parts of premix and 1-3.5 parts of nitrate in parts by mass.
For use as described above, the premix preferably provides 25mg of iron (Fe), 40mg of manganese (Mn), 40mg of zinc (Zn), 8mg of copper (Cu), 0.3mg of iodine (I), 0.2mg of selenium (Se), 0.1mg of cobalt (Co), 940IU of Vitamin A (VA), 111IU of Vitamin D (VD), and 20IU of Vitamin E (VE) per kg of diet.
The feed comprises, by mass, 17.76 parts of corn straws, 10.34 parts of soybean hulls, 2.00 parts of sunflower hulls, 48.60 parts of corns, 11.35 parts of soybean meal, 3.00 parts of sesame cakes, 5.00 parts of molasses, 0.25 part of stone powder, 0.70 part of NaCl, 1.00 part of premix and 3 parts of nitrate.
For the above-mentioned applications, preferably, the nitrate is calcium nitrate.
The invention has the beneficial effects that:
according to the method provided by the invention, calcium nitrate is added into the feed to feed the Hu sheep, so that the rumen nipple width, the thickness of a muscular layer and the width of ileum villus of the Hu sheep are obviously increased, the thickness of abomasum mucous membrane, the length of jejunum villus, the width of villus and the villus hiding ratio are obviously reduced, the calcium nitrate has certain influence on the gastrointestinal tract tissue morphological development of the Hu sheep, the rumen and ileum tissue morphological development is obviously improved, and the nutrient absorption utilization rate is improved; the weight of the Hu sheep carcass, head, hoof and internal organs can be effectively increased; obviously reduces the methanococcus duodenale and colon, obviously reduces the methanobrevibacterium colocolinum and has the tendency of reducing the methanobrevibacterium rumen, and shows that the calcium nitrate can inhibit the methane generation in the gastrointestinal tract of the Hu sheep and is beneficial to environmental protection. Although calcium nitrate inhibits rumen fermentation of Hu sheep to reduce fiber decomposition capacity, calcium nitrate improves the abundance of Hu sheep colon vibrio succinogenes to improve the decomposition and utilization efficiency of fiber substances in colon, and can be better absorbed and utilized for diet.
Drawings
FIG. 1 is a dilution curve of species diversity.
FIG. 2 is a bar graph of the relative abundance of species at the level of the gastroenterology gate.
Fig. 3 is a graph of the principal coordinate analysis of the rumen microflora.
FIG. 4 is a diagram showing the principal coordinate analysis of the microbial flora of duodenum.
FIG. 5 is a diagram of the principal coordinates of the colon microbial flora.
FIG. 6 is a UPGMA clustering tree and a species relative abundance distribution plot at the phylum level for each sample.
Detailed Description
The following examples are intended to further illustrate the invention but should not be construed as limiting it. Modifications and substitutions may be made thereto without departing from the spirit and scope of the invention.
Unless otherwise indicated, the technical means used in the examples are conventional means well known to those skilled in the art, and unless otherwise specified, the reagents used in the methods are analytically pure or above.
Example 1
Selecting 30 male Hu sheep lambs which are healthy, have good growth and development and have close weight to 120 days old for a growth fattening test, and randomly dividing the selected Hu sheep into 2 groups according to the principle that the weights of the groups are similar, wherein each group comprises 15 lambs, and each lambs is a repetition. The control group of Hu sheep was fed basal diet, and the test group of Hu sheep was fed diet containing 3% calcium nitrate. And (4) slaughtering the Hu sheep at a transition period of 15d and a positive test period of 60d when the Hu sheep is raised to 61 d.
Before selecting test sheep, the sheep house used in the test is disinfected in all directions by using powerful disinfectant, and the disinfected sheep house allows disinfectant in the sheep house to volatilize for a period of time and then enters the sheep so as to avoid discomfort of the sheep. After the test formally begins, the test is fed in a single cage, and the test is fed with the feed once every morning and afternoon, so that the test can be fed freely and can drink water freely. The test is completed in Defu agriculture science and technology limited company in civil service county of Gansu province. The used feed is produced and processed by Gansu Runming bioengineering Limited liability company, and the composition (in parts by mass) and the nutritional ingredients of the feed are shown in Table 1.
TABLE 1 feed composition and nutrient composition table (Dry matter basis)
Wherein, the premix is provided for each kg of diet: 25mg of iron (Fe), 40mg of manganese (Mn), 40mg of zinc (Zn), 8mg of copper (Cu), 0.3mg of iodine (I), 0.2mg of selenium (Se), 0.1mg of cobalt (Co), 940IU of Vitamin A (VA), 111IU of Vitamin D (VD), and 20IU of Vitamin E (VE).
Firstly, collecting a sample:
1. tissue and organ sample
After the test sheep is slaughtered, the head, hooves and internal organs are weighed in sequence, and data are recorded. And measuring the size of the sheep skin and recording data. And the ratio of the live weight to the weight of the live weight before slaughter was calculated, and the results are shown in Table 1.
TABLE 1 influence of calcium nitrate addition to diets on the slaughter performance and tissue and organs of Hu sheep
It can be seen that the addition of calcium nitrate to the diet has an effect of improving the slaughtering performance and tissues and organs of the Hu sheep, but is not significant (P is more than 0.05).
2. Gastrointestinal tract sample
After the test sheep is slaughtered, the gastrointestinal tract is separated, different parts of the gastrointestinal tract are weighed after separation, the weight is recorded, and the length of each intestinal section is measured. ComputingDifferent stomach weights account for the total stomach weight ratio, different intestine sections account for the total intestine section weight ratio, and different intestine section lengths account for the total intestine section length ratio. Meanwhile, cutting the rumen, the duodenum and the colon by using scissors to enable the contents to leak out, collecting rumen, duodenum and colon chyme by using a sterilized cryopreservation tube, quickly freezing the cryopreservation tube filled with gastrointestinal chyme in liquid nitrogen, and transferring to a refrigerator at minus 80 ℃ for storage for subsequent microbial determination. Then using scissors to respectively cut about 2cm from the rumen, the duodenum and the middle part of the colon2And (3) quickly placing the tissue in 4% paraformaldehyde for fixation, and using the tissue for subsequent gastrointestinal tract tissue morphology section preparation and observation.
Second, test methods and results
Preparation and observation of gastrointestinal tract tissue slices: the collected gastrointestinal tissues are sent to Changsha Zhifan biotechnology Limited liability company in Hunan to be sliced by a hematoxylin-eosin staining method. The sections were observed using an Olympus-DP71 microscope, and the thickness of the muscular layer of the intestinal tissue, the depth of the crypt, and the length and width of the villus were counted, and the hiding ratio of the villus was calculated. The results of counting the length, width, thickness of muscular layer and thickness of epithelial cell of rumen tissue are shown in Table 2.
TABLE 2 Effect of calcium nitrate addition to diets on the relative quality and relative length of the Hu sheep gastrointestinal tract
The results show that the addition of calcium nitrate in the diet significantly reduces the weight of the cecum/total intestinal tract of the Hu sheep (P < 0.05), and has no significant influence on the relative quality and relative length of other gastrointestinal tracts.
TABLE 3 influence of calcium nitrate addition to diets on Hu sheep gastrointestinal morphology
The results in Table 3 show that the addition of calcium nitrate to the feed significantly increases the rumen papilla width, the thickness of the muscular layer and the width of the ileal villus of the Hu sheep (P < 0.05), significantly reduces the thickness of the abomasum mucosa, the length of the jejunum villus, the width of the villus and the villus hiding ratio (P < 0.05), and has no significant influence on the shapes of other gastrointestinal tract tissues (P > 0.05). This shows that calcium nitrate has certain effect on the morphological development of gastrointestinal tract tissues of Hu sheep and has an improvement effect on the morphological development of rumen and ileum tissues.
Third, determination of gastrointestinal tract microbial diversity
1 determination of microbial diversity
The collected rumen chyme, colon chyme and duodenum chyme are sent to Beijing Nuo He Zhi Yuan science and technology Co., Ltd, and sequencing is carried out by utilizing Illumina NovaSeq platform. The main operation flow is as follows: extracting total genomic DNA of the chyme sample by adopting a CTAB method. The extracted genomic DNA was then monitored for concentration and purity using a 1% agarose gel, and the results are shown in FIG. 1. In the figure, sr.a (duodenum control group), sr.c (duodenum calcium nitrate group), jc.a (colon control group), jc.c (colon calcium nitrate group), lw.a (rumen control group), lw.c (rumen calcium nitrate group). The result shows that the dilution curves of all samples tend to be flat, which indicates that the sequencing data amount is gradual and reasonable, and can well reflect most of microorganism conditions in gastrointestinal tract samples.
After the DNA quality is qualified, a specific primer 16S V3-V4 is used
The 16S rRNA of the genomic DNA of the sample was amplified using the High-Fidelity PCR Master Mix with GC Buffer kit (NEW England Biolabs). The amplified PCR products were mixed with the same volume of 1X carrier fluid (containing SYB green) and then detected by electrophoresis on a 2% agarose gel. The PCR products are mixed according to equal density proportion. The mixture PCR products were then purified using a Qiagen gel extraction kit (Qiagen, germany). Then use
The DNA PCR-Free sample preparation kit (Illumina, USA) generates a sequencing library and adds an index code. Library quality was assessed on a Qubit @2.0 fluorometer (Thermo Scientific) and Agilent Bioanalyzer 2100 system. And finally, sequencing the library on an Illumina NovaSeq platform to generate the paired end reads of 250 bp.
1,285,763 original off-machine sequences are obtained by rumen through Illumina NovaSeq sequencing, 799,104 effective sequences for analysis are remained after splicing and filtering, each sample contains 49,944 effective sequences on average, and the length of each sequence is 417bp on average. 1,697,211 original off-line sequences are obtained from the colon, 1,043,921 effective sequences for analysis are remained after splicing and filtering, each sample contains 65,245 effective sequences on average, and the length of each sequence is 413bp on average. 1,695,420 original off-machine sequences are obtained in duodenum, 1,045,420 effective sequences for analysis are remained after splicing and filtering, each sample contains 65,338 effective sequences on average, and the length of each sequence is 410bp on average. Common and specific microorganisms between the control and calcium nitrate groups were analyzed based on clustering results and study requirements for OTUs. At 97% similarity level, a total of 2248 OUTs were produced in the duodenal chyme sample, of which 1848 in the control group, 1951 in the calcium nitrate group, and 1551 in the 2 groups. A total of 2554 OUTs were produced in the colonic chyme sample, with 2180 in the control group, 1798 in the calcium nitrate group and 1424 shared by the 2 groups. A total of 3149 OUTs were produced in the rumen chyme sample, with 2344 in the control group, 2429 in the calcium nitrate group and 1624 shared by 2 groups.
2 analysis of biological information
Firstly, carrying out FLASH to merge and splice paired end readings of an original sequence obtained after sequencing; and then, carrying out quality filtration on the original sequence under a specific filtration condition, comparing the sequence with a reference database, detecting a chimera sequence, removing chimeras and finally obtaining an effective sequence. Sequences were analyzed using Upearse software and sequences with > 97% similarity were assigned to the same OUT. For each representative sequence, classification information was annotated using a Silva database. To study the phylogenetic relationship of different OTUs and the difference of dominant species in different samples (groups), multiple sequence alignments were performed using the MUSCLE software. The abundance information of OTUs was normalized using the sequence number standard corresponding to the least sequenced sample. Alpha and Beta diversity were then analyzed based on this output normalized data. Alpha diversity was shown by R software (version 2.15.3) using QIIME (version 1.7.0) software calculations. Beta diversity for weighted and unweighted unifrc was calculated using QIIME software (version 1.9.1). And according to the species annotation result, selecting the top 10 highest abundance species of each group on the gate classification level, generating a column accumulation graph of the relative abundance of the species, and calculating the difference significance among the groups. And (4) a main coordinate analysis chart of the gastrointestinal tract microbial community drawn based on Weighted Unifrac distance. And performing phylogenetic clustering analysis on different gastrointestinal tract microorganisms, and drawing a sample clustering tree of species composition distribution with the maximum relative abundance ranking as the top ten, wherein the result is as follows:
alpha diversity analysis
At the 97% consistency threshold, the Alpha diversity index (shannon, simpson, chao1, ACE, goods _ coverage) of different gastrointestinal tracts of the hu sheep is shown in table 4.
TABLE 4 influence of calcium nitrate addition to diets on Hu sheep gastrointestinal tract microbial Alpha diversity index
As can be seen from Table 4, the addition of calcium nitrate to the diet significantly reduced the chao1 index and ACE index (P < 0.05) in the Hu sheep colonic chyme, significantly increased the coverage in the colonic chyme (P < 0.05), and had no significant effect on the other gastrointestinal tract Alpha diversity index (P > 0.05). Indicating that calcium nitrate reduced the total number of microbial species and the number of OUT in the colony in the colon of Hu sheep. In the invention, 9741 OUT are obtained in the colon of the Hu sheep of the control group, and 1271 OUT are obtained in each sample on average; the calcium nitrate group yielded 8560 OUT's, with an average of 1070 OUT's per sample. The rumen of the Hu sheep of the control group is separated by 6295 OUT, and 787 OUT are averagely obtained in each sample; the calcium nitrate group yielded 5899 OUT, with an average of 737 OUT per sample. The number of OUT is higher than that of Chengxiang nitrate, the influence of nitrate on the disappearance rate of nitrate nitrogen in the rumen of Hu sheep, the fermentation characteristics and the microflora [ D ], the average detection result of 515OUT per sample of the Yangzhou university of Jiangzhou, 2016, is much higher than that of Pandya P, Singh K, Parnerkar S, et al. This shows that the invention has more abundant gastrointestinal tract microorganism information for research, and the research result is more representative.
Beta diversity analysis
A histogram of the relative abundance of species at the level of the gastrointestinal tract portal is shown in FIG. 2, in which: sr.a (duodenal control group), sr.c (duodenal calcium nitrate group), jc.a (colonic control group), jc.c (colonic calcium nitrate group), lw.a (rumen control group), lw.c (rumen calcium nitrate group). The effect of calcium nitrate addition to the diet on the abundance of microbial species in the gastrointestinal tract at the level of the phylum Hu sheep is shown in Table 5.
TABLE 5 Effect of calcium nitrate addition to diets on the abundance of microbial species in the gastrointestinal tract at the level of the phylum Hu sheep
As can be seen from fig. 2 and table 5, the relative abundance of bacteroidetes (bacteroideta), actinomycetes (actinobacteroideta), Firmicutes (Firmicutes), Proteobacteria (Proteobacteria) and eurycota (euryarchaeotada) in the gastrointestinal tract of different groups of hu sheep is about 90%. Wherein, the dominant bacteria in the rumen are bacteroidetes (Bacteroidota) and Firmicutes (Firmicutes), which respectively account for 47.40 percent and 28.65 percent; the dominant bacteria in the colon are bacteroidetes (Bacteroidota) and Firmicutes (Firmicutes), which respectively account for 21.92% and 57.34%; the dominant bacteria in the duodenum are the phylum actinomycetemcomita, Firmicutes and euryarchaeotid (actinobacillus), accounting for 22.96%, 53.60% and 13.14%, respectively. In addition, the microbial flora composition in the gastrointestinal tract of the Hu sheep was also very different for the different treatments.
As can be seen from Table 5, the addition of calcium nitrate to the diet significantly reduced the abundance of Cellobiotomycota (Fibromobacterota), Actinomycetes (Actionbacteriota) and eurycota (Euryarchaeotid) in the rumen of Hu sheep (P < 0.05), significantly increased the abundance of Proteobacteria (Proteobacteria) and Others (P < 0.05) in the duodenum, and had no significant effect on the abundance of other gastrointestinal microbial species (P > 0.05).
The principal coordinate analysis plots of the gastrointestinal microflora are shown in FIGS. 3-5, in which SR.A (samples of duodenum control group), SR.C (samples of calcium nitrate group of duodenum), JC.A (samples of colon control group), JC.C (samples of calcium nitrate group of colon), LW.A (samples of rumen control group), LW.C (samples of calcium nitrate group of rumen). In fig. 3, it can be seen that when the contribution rate of PCoA1 is 35.71%, and the contribution rate of PCoA2 is 22.87%, the distance between the calcium nitrate group and the control group of the ruminal sample of the hu sheep is relatively short, indicating that the similarity of the structural composition of the ruminal microflora of the hu sheep between the two treatment groups is relatively high. The distribution of the calcium nitrate group Hu sheep rumen samples is relatively discrete, while the comparison of the calcium nitrate group Hu sheep rumen samples is centralized, which indicates that the structural similarity of rumen microorganisms among individuals in the calcium nitrate group is lower than that among individuals in the control group. As can be seen from fig. 4, when the contribution rate of PCoA1 was 38.75% and the contribution rate of PCoA2 was 22.51%, the calcium nitrate group and the control group of the sample of the duodenum of the hu sheep were distinguished, indicating that the structural similarity of the microflora of the hu sheep was low between the two treatment groups. As can be seen from fig. 5, when the contribution rate of PCoA1 was 45.91% and the contribution rate of PCoA2 was 12.87%, the calcium nitrate group and the control group of the hu sheep colon samples were clearly distinguished, indicating that the structural similarity of the hu sheep colon microbial community between the two treatment groups was low.
The results of the effect of calcium nitrate on the abundance of methanogens in the gastrointestinal tract at the level of Hu sheep are shown in Table 6, and it can be seen from the results in the table that the addition of calcium nitrate to the diet significantly reduces the abundance of Brevibacterium methanolicum (Methanobacter) and Methanosphaericus Methanosphaera (Methanosphaera) in the colon of Hu sheep (P < 0.05), and significantly reduces the abundance of Methanosphaera Methanosphaera (P < 0.05).
TABLE 6 Effect of calcium nitrate addition to diets on Hu sheep level gastrointestinal methanogen abundance
The results of the effect of calcium nitrate addition to the diet on Hu sheep level gastrointestinal tract cellulolytic bacteria are shown in Table 7. It can be seen that the addition of calcium nitrate to the diet significantly reduced the abundance of the Hu sheep rumen filamentous bacterium (Fibrobacter) (P < 0.05), with no significant effect on the fibrolytic bacteria at other GI levels (P > 0.05).
TABLE 7 Effect of calcium nitrate addition to diets on Hu sheep-level gastrointestinal fibrolytic bacteria
The results show that calcium nitrate has an increasing tendency towards Vibrio succinogenes in the Hu sheep colon (P < 0.10), which indicates that calcium nitrate can increase the fiber-decomposing capacity of the Hu sheep colon. The cellulolytic bacteria produce hydrogen gas when decomposing fibers, and the reduction process of nitrate needs to consume hydrogen, thereby promoting the rapid decomposition of the cellulolytic bacteria and enhancing the decomposition capability thereof.
The relative abundance distribution of species at the phylum level for the UPGMA cluster tree and each sample is shown in figure 6. The left side of the graph is the clustering tree structure and the right side is the distribution graph of the relative abundance of species at the portal level for each group of samples, wherein sr.a (duodenal control group), sr.c (duodenal calcium nitrate group), jc.a (colonic control group), jc.c (colonic calcium nitrate group), lw.a (rumen control group), lw.c (rumen calcium nitrate group). Firmicutes, bacteroidetes (bacteroidata), Proteobacteria (Proteobacteria), unidentified Bacteria (unidentified _ Bacteria), actinomycetes (Actinobacteriota), spirochetes (spirochaeta), acidobacteria (acidobacteria), Verrucomicrobiota, clodotomyelia (Chloroflexi), eurycota (euryarchaeta), other Bacteria atheros. It can be seen that the different gastrointestinal tract microbial organisms of the rumen, colon and duodenum of the Hu sheep are differentiated, which indicates that the different gastrointestinal tract microbial compositions are different. The microbial similarity of the colon of Hu sheep in the control group and the calcium nitrate group is low, and two clusters are obviously formed, which indicates that the calcium nitrate has a large influence on the colon microbial composition. After calcium nitrate is added into the feed, the similarity of the microbial composition in the rumen and the duodenum of the Hu sheep is higher, which shows that the calcium nitrate has less influence on the microbial composition in the rumen and the duodenum of the Hu sheep.
From the above, it can be seen that the addition of calcium nitrate to the diet improves the gastrointestinal microbial composition of the Hu sheep. The regulation and control of the Hu sheep gastrointestinal tract microflora by the calcium nitrate can effectively reduce the Hu sheep gastrointestinal tract methane emission and realize the environment-friendly Hu sheep feeding. In addition, the digestion, absorption and utilization efficiency of fibrous substances in the Hu sheep colon is improved by regulating and controlling the Hu sheep colon microflora.
Compared with the effect of feed additives or methane inhibitors used in feeding other ruminants, the research of the invention finds that the use of a proper amount of calcium nitrate in the feeding of the Hu sheep effectively reduces the discharge of methane in the gastrointestinal tract of the Hu sheep by inhibiting the abundance of gastrointestinal methanogenic archaea on the premise of not influencing the production performance of the Hu sheep, and is the best choice for building environment-friendly and sustainable development of the Hu sheep breeding industry at present.
Claims (6)
1. The nitrate is applied to improving the morphological development of the gastrointestinal tract tissue of the Hu sheep and improving the microorganism.
2. The use according to claim 1, wherein the nitrate is present in an amount of 1-3% of the feed.
3. The application of the feed as claimed in claim 1, wherein the feed comprises 15-18 parts of corn straw, 9-11 parts of soybean hull, 1-3 parts of sunflower hull, 45-50 parts of corn, 10-12 parts of soybean meal, 2-4 parts of sesame cake, 3-6 parts of molasses, 0.2-0.5 part of stone powder, 0.60-0.8 part of NaCl0.5-1.50 parts of premix and 1-3.5 parts of nitrate by mass.
4. The use according to claim 3 wherein the premix provides 25mg of iron, 40mg of manganese, 40mg of zinc, 8mg of copper, 0.3mg of iodine, 0.2mg of selenium, 0.1mg of cobalt, 940IU of vitamin A, 111IU of vitamin D, 20IU of vitamin E per kg of diet.
5. The use according to claim 1, characterized in that the feed comprises, by mass, 17.76 parts of corn stalks, 10.34 parts of soybean hulls, 2.00 parts of sunflower hulls, 48.60 parts of corns, 11.35 parts of soybean meal, 3.00 parts of sesame cakes, 5.00 parts of molasses, 0.25 part of stone powder, 0.70 part of NaCl, 1.00 part of premix and 3 parts of nitrate.
6. Use according to claim 1, characterized in that the nitrate is calcium nitrate.
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| 陈志远;马婷婷;方伟;左晓昕;林淼;赵国琦;: "日粮硝酸盐水平对湖羊瘤胃硝态氮动态消失率、发酵参数及血液高铁血红蛋白含量的影响" * |
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| CN115462447A (en) * | 2022-10-06 | 2022-12-13 | 中国科学院亚热带农业生态研究所 | Method for preparing sorghum straw whole-plant corn mixed storage forage grass |
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