Traditional Chinese and Tibetan medicine compound preparation for promoting postpartum recovery of yaks and shortening estrus cycle as well as preparation method and application of traditional Chinese and Tibetan medicine compound preparation

Abstract

The invention discloses a Chinese and Tibetan medicine compound preparation for promoting postpartum recovery of yaks and shortening the estrus cycle as well as a preparation method and application thereof, and belongs to the technical field of Chinese and Tibetan medicines. The traditional Chinese and Tibetan medicine compound preparation is prepared from the following raw materials in parts by weight: 15-20 parts of scutellaria baicalensis, 15-20 parts of Tibetan codonopsis pilosula, 15-20 parts of astragalus membranaceus, 15-20 parts of radix paeoniae alba, 5-10 parts of Tibetan rhizoma polygonati and 5-10 parts of liquorice. After the Chinese and Tibetan medicine compound preparation is applied to postpartum female yaks, the physique of the postpartum female yaks can be enhanced, the oxidation resistance of the postpartum female yaks can be improved, and therefore the effects that the body functions of the postpartum female yaks are recovered, immunity is enhanced, the hormone level is improved, and the estrus cycle is shortened are achieved; the problem of low breeding efficiency caused by insufficient nutrition and severe environment in the traditional breeding process is effectively solved.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese and Tibetan medicines, and particularly relates to a traditional Chinese and Tibetan medicine compound preparation for promoting the postpartum recovery of yaks and shortening the estrus cycle, and a preparation method and application thereof. Background Art

[0002] Yaks mainly inhabit the Qinghai-Tibet Plateau and its surrounding areas in China, and are one of the important livestock species indispensable to the animal husbandry in this region.

[0003] However, the reproductive characteristics of yaks pose challenges to their breeding efficiency. Yaks have obvious estrus seasonality, usually concentrated from June to September, and the gestation period is as long as 259 days. This results in the entire pregnancy period being in the cold season. Especially in the later stage of rapid fetal development, female yaks face serious nutritional deficiencies, which in turn affect the maternal health and subsequent reproductive ability, and also weaken the resistance of newborn yaks. In addition, the traditional "mother-calf" nursing mode requires female yaks to consume a large amount of nutrients for lactation after giving birth, further delaying the process of their physical recovery and the restoration of the reproductive system, resulting in an inefficient breeding situation of "one calf every two years" or even "one calf every three years" for most yaks, seriously restricting the sustainable development of the yak industry.

[0004] Therefore, developing a drug that is both efficient and safe, does not cause drug resistance, and can improve the physique of female yaks, enhance reproductive performance, promote postpartum recovery, strengthen immunity and shorten the estrus cycle has become one of the urgent problems to be solved by those skilled in the art. Summary of the Invention

[0005] In view of the above deficiencies of the prior art, the present invention discloses a traditional Chinese and Tibetan medicine compound preparation for promoting the postpartum recovery of yaks and shortening the estrus cycle, and a preparation method and application thereof. After the traditional Chinese and Tibetan medicine compound preparation disclosed by the present invention is applied to postpartum female yaks, it can enhance the physique of postpartum female yaks, significantly improve the antioxidant capacity of postpartum female yaks, etc., so as to achieve the effects of restoring the body functions of postpartum female yaks, enhancing immunity, increasing hormone levels to shorten the estrus cycle, etc., and effectively solve the problem of low breeding efficiency caused by nutritional deficiencies and harsh environments in the traditional breeding process.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] The present invention discloses a traditional Chinese and Tibetan medicine compound preparation for promoting the postpartum recovery of yaks and shortening the estrus cycle. The traditional Chinese and Tibetan medicine compound preparation is made from the following raw materials in parts by weight: 15-20 parts of Scutellaria baicalensis Georgi, 15-20 parts of Codonopsis thalictrifolia Wall., 15-20 parts of Astragalus membranaceus (Fisch.) Bunge, 15-20 parts of Paeonia lactiflora Pall., 5-10 parts of Polygonatum cirrhifolium (Wall.) Royle, and 5-10 parts of Glycyrrhiza uralensis Fisch.

[0008] Preferably, the traditional Chinese and Tibetan medicine compound preparation is made from the following raw materials in parts by weight: 18 parts of Scutellaria baicalensis, 18 parts of Codonopsis tangshen, 18 parts of Astragalus membranaceus, 18 parts of Paeonia lactiflora, 7 parts of Polygonatum sibiricum, and 7 parts of Glycyrrhiza uralensis.

[0009] The present invention provides a preparation method of the above-mentioned traditional Chinese and Tibetan medicine compound preparation for promoting the postpartum recovery of yaks and shortening the estrus cycle, comprising the following steps:

[0010] S1. Weigh appropriate amounts of Scutellaria baicalensis, Codonopsis tangshen, Astragalus membranaceus, Paeonia lactiflora, Polygonatum sibiricum, and Glycyrrhiza uralensis respectively according to parts by weight. Each medicinal material is dried one by one, and then the dried medicinal materials are respectively pulverized to obtain medicinal material powders.

[0011] S2. The pulverized medicinal material powders in step S1 are respectively screened through a sieve, and then the screened medicinal material powders of each kind are mixed evenly according to the above-mentioned parts by weight to obtain the traditional Chinese and Tibetan medicine compound preparation.

[0012] Preferably, in step S2, the screened medicinal material powders are mixed according to parts by weight as follows: 15 - 20 parts of Scutellaria baicalensis, 15 - 20 parts of Codonopsis tangshen, 15 - 20 parts of Astragalus membranaceus, 15 - 20 parts of Paeonia lactiflora, 5 - 10 parts of Polygonatum sibiricum, and 5 - 10 parts of Glycyrrhiza uralensis.

[0013] Preferably, in step S2, the mesh number of the sieve is 30 - 50 meshes.

[0014] The present invention also discloses the application of the above-mentioned traditional Chinese and Tibetan medicine compound preparation in the preparation of drugs for promoting the postpartum recovery of yaks or shortening the estrus cycle.

[0015] Preferably, the application of the traditional Chinese and Tibetan medicine compound preparation in the preparation of drugs or feed additives for postpartum weaned female yaks.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] (1) The traditional Chinese and Tibetan medicine compound preparation provided by the present invention is added to the feed for supplementary feeding of postpartum weaned female yaks. Feeding the feed containing the traditional Chinese and Tibetan medicine compound preparation can significantly improve the antioxidant capacity and regulate the inflammatory level of postpartum weaned female yaks, and at the same time have a positive impact on the rumen microbial diversity and the composition structure of the flora, promoting the postpartum female yaks to recover their body functions, enhance their physique and immunity.

[0018] (2) The traditional Chinese and Tibetan medicine compound preparation provided by the present invention reduces harmful bacteria by enriching beneficial bacteria, effectively regulates the composition structure of the intestinal flora, promotes beneficial changes in the intestinal flora of postpartum female yaks, especially increases the relative abundance of beneficial bacteria and reduces the relative abundance of harmful bacteria. After being applied to postpartum female yaks, it can effectively restore the body functions of female yaks and enhance their immunity, providing a new method and reference basis for the breeding of postpartum weaned female yaks in the plateau area.

[0019] (3) The traditional Chinese and Tibetan medicine compound preparation provided by the present invention can increase the levels of estrogen, luteinizing hormone, and follicle-stimulating hormone in postpartum female yaks, thereby helping to shorten the estrus cycle of female yaks, improving the reproductive efficiency of yaks, and promoting the sustainable development of the yak industry in the Qinghai-Tibet Plateau region. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Detection of serum indexes of postpartum female yaks by the traditional Chinese and Tibetan medicine compound preparation provided by the present invention;

[0021] Figure 2 Analysis of the alpha diversity of rumen microorganisms of postpartum female yaks by the traditional Chinese and Tibetan medicine compound preparation provided by the present invention;

[0022] Figure 3 Venn diagram of rumen microorganisms of postpartum female yaks by the traditional Chinese and Tibetan medicine compound preparation provided by the present invention;

[0023] Figure 4 Analysis of the beta diversity of rumen microorganisms of postpartum female yaks by the traditional Chinese and Tibetan medicine compound preparation provided by the present invention;

[0024] Figure 5 Effect of the traditional Chinese and Tibetan medicine compound preparation provided by the present invention on rumen microorganisms of postpartum female yaks at different taxonomic levels;

[0025] Figure 6 LEfSe analysis of rumen microorganisms of postpartum female yaks by the traditional Chinese and Tibetan medicine compound preparation provided by the present invention;

[0026] Figure 7 Analysis of the alpha diversity of intestinal flora of postpartum female yaks by the traditional Chinese and Tibetan medicine compound preparation provided by the present invention;

[0027] Figure 8 Venn diagram of intestinal flora of postpartum female yaks by the traditional Chinese and Tibetan medicine compound preparation provided by the present invention;

[0028] Figure 9 Analysis of the beta diversity of intestinal flora of postpartum female yaks by the traditional Chinese and Tibetan medicine compound preparation provided by the present invention;

[0029] Figure 10 Effect of the traditional Chinese and Tibetan medicine compound preparation provided by the present invention on intestinal flora of postpartum female yaks;

[0030] Figure 11 LEfSe analysis of intestinal flora of postpartum female yaks by the traditional Chinese and Tibetan medicine compound preparation provided by the present invention;

[0031] Figure 12 Effect of the traditional Chinese and Tibetan medicine compound preparation provided by the present invention on the hormone levels of postpartum female yaks;

[0032] Figure 13 Statistical results of the time of the second estrus of postpartum female yaks for the traditional Chinese and Tibetan medicine compound preparation provided by the present invention. Specific embodiments

[0033] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0034] The experimental methods used in the following embodiments are all conventional methods unless otherwise specified.

[0035] The materials, reagents, etc. used in the following embodiments can be obtained from commercial channels unless otherwise specified.

[0036] The present invention discloses a traditional Chinese and Tibetan medicine compound preparation for promoting the postpartum recovery of yaks and shortening the estrus cycle. The traditional Chinese and Tibetan medicine compound preparation is made from the following raw materials in parts by weight: 15-20 parts of Scutellaria baicalensis Georgi, 15-20 parts of Codonopsis pilosula Franch. var. thalictrifolia (Wall.) Tsai & Tseng, 15-20 parts of Astragalus membranaceus (Fisch.) Bunge, 15-20 parts of Paeonia lactiflora Pall., 5-10 parts of Polygonatum cirrhifolium (Wall.) Royle, 5-10 parts of Glycyrrhiza uralensis Fisch.

[0037] The above traditional Chinese and Tibetan medicine compound preparation is most preferably made from the following raw materials in parts by weight: 18 parts of Scutellaria baicalensis Georgi, 18 parts of Codonopsis pilosula Franch. var. thalictrifolia (Wall.) Tsai & Tseng, 18 parts of Astragalus membranaceus (Fisch.) Bunge, 18 parts of Paeonia lactiflora Pall., 7 parts of Polygonatum cirrhifolium (Wall.) Royle, 7 parts of Glycyrrhiza uralensis Fisch.

[0038] The present invention provides a preparation method of the above traditional Chinese and Tibetan medicine compound preparation for promoting the postpartum recovery of yaks and shortening the estrus cycle, including the following steps:

[0039] S1. Weigh 15-20 parts of Scutellaria baicalensis Georgi, 15-20 parts of Codonopsis pilosula Franch. var. thalictrifolia (Wall.) Tsai & Tseng, 15-20 parts of Astragalus membranaceus (Fisch.) Bunge, 15-20 parts of Paeonia lactiflora Pall., 5-10 parts of Polygonatum cirrhifolium (Wall.) Royle, and 5-10 parts of Glycyrrhiza uralensis Fisch. by weight. Each medicinal material is dried one by one, and then the dried medicinal materials are respectively pulverized to obtain medicinal material powders.

[0040] S2. The medicinal material powders pulverized in step S1 are respectively screened through a sieve with a mesh size of 30-50 meshes, and then each screened medicinal material powder is mixed evenly according to the above weight parts to obtain the traditional Chinese and Tibetan medicine compound preparation.

[0041] Among them, the screened medicinal material powders are mixed in parts by weight as 15-20 parts of Scutellaria baicalensis Georgi, 15-20 parts of Codonopsis pilosula Franch. var. thalictrifolia (Wall.) Tsai & Tseng, 15-20 parts of Astragalus membranaceus (Fisch.) Bunge, 15-20 parts of Paeonia lactiflora Pall., 5-10 parts of Polygonatum cirrhifolium (Wall.) Royle, and 5-10 parts of Glycyrrhiza uralensis Fisch.

[0042] The present invention also discloses the application of the above-mentioned traditional Chinese and Tibetan medicine compound preparation in the preparation of a drug for promoting the postpartum recovery of yaks or shortening the estrus cycle.

[0043] Among them, the application of the above-mentioned Tibetan medicine compound preparation in the preparation of a drug or feed additive for postpartum weaned female yaks.

[0044] The technical solution of the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments:

[0045] Example 1

[0046] In this example, the preparation method of the traditional Chinese and Tibetan medicine compound preparation for promoting the postpartum recovery of yaks and shortening the estrus cycle is as follows:

[0047] S1. By weight, 18 parts of Scutellaria baicalensis, 18 parts of Codonopsis thalictrifolia Wall., 18 parts of Astragalus membranaceus, 18 parts of Paeonia lactiflora, 7 parts of Polygonatum cirrhifolium (Wall.) Royle, and 7 parts of Glycyrrhiza uralensis Fisch. are weighed respectively. Each medicinal material is dried one by one, and then the dried medicinal materials are respectively pulverized to obtain medicinal material powders.

[0048] S2. The medicinal material powders obtained by pulverization in step S1 are respectively screened through a 30-mesh sieve, and then each screened medicinal material powder is mixed evenly according to 18 parts of Scutellaria baicalensis, 18 parts of Codonopsis thalictrifolia Wall., 18 parts of Astragalus membranaceus, 18 parts of Paeonia lactiflora, 7 parts of Polygonatum cirrhifolium (Wall.) Royle, and 7 parts of Glycyrrhiza uralensis Fisch., thus preparing the traditional Chinese and Tibetan medicine compound preparation.

[0049] Example 2

[0050] This example is about the effects of the traditional Chinese and Tibetan medicine compound preparation prepared in Example 1 on the serum indexes and rumen microbial diversity of postpartum female yaks.

[0051] Twenty postpartum weaned female yaks 80 ± 15 days after parturition were selected for the experiment and randomly divided into a control group (DH) and a traditional Chinese and Tibetan medicine compound preparation supplementary feeding group (DE), with 10 heads in each group. The traditional Chinese and Tibetan medicine compound preparation supplementary feeding group (DE) was fed a basal diet (5 kg) supplemented with 5% of the traditional Chinese and Tibetan medicine compound preparation prepared in Example 1 every day, while the blank control group (DH group) was fed 5 kg of basal diet every day. The environmental conditions were kept consistent in all groups. The first 7 days before the start of the experiment were the adaptation period. From the 1st to the 4th day, 1 / 4 of the experimental dose of the traditional Chinese and Tibetan medicine compound preparation was given, and from the 5th to the 7th day, 1 / 2 of the experimental dose was given to observe whether there were any adverse reactions in the postpartum female yaks; from the 8th day, the experiment period officially began, and the dosing dose was adjusted to 5% of the traditional Chinese and Tibetan medicine compound preparation. The experiment lasted for 30 days; the daily feeding time was 9:30 and 15:00 to ensure that the traditional Chinese and Tibetan medicine compound preparation was fully mixed with the basal diet, and the female yaks drank water freely during the experiment.

[0052] (ⅰ) On the 15th and 30th days of the experiment, blood samples of all female yaks were collected by jugular venipuncture. After standing at room temperature for 2 - 3 h, the serum was separated by centrifugation at 3500 r / min for 15 min. The serum was used to detect the indexes related to the immune capacity and antioxidant capacity of female yaks, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10), malondialdehyde (MDA), total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD).

[0053] (ⅱ) Before morning feeding on the 15th and 30th days of the experiment, 50 mL of rumen fluid was drawn from the rumen of each female yak in the DE group and the DH group respectively. The rumen fluid samples on the 15th day were classified as DE1 (divided into DA1 - DA6 between groups) and DH1 (divided into DD1 - DD6 between groups), and the samples on the 30th day were classified as DE2 (divided into DA7 - DA12 between groups) and DH2 (divided into DD7 - DD12 between groups). After being filtered through four layers of sterilized gauze, the rumen fluid was sub-packed for subsequent 16S rRNA sequencing analysis to evaluate the changes in rumen microbial diversity. Through the above experimental design, the comprehensive effects of the traditional Chinese and Tibetan medicine compound preparation of the present invention on the physiology and microbial community of female yaks were explored, and the specific analysis is as follows.

[0054] (1) Figure 1For the detection of the serum indexes of postpartum female yaks on the 15th and 30th days of the experiment for the prepared traditional Chinese and Tibetan medicine compound preparation, it can be seen from the figure that: (i) On the 15th and 30th days of the experiment, the superoxide dismutase (SOD) activity of the DE group was significantly higher than that of the DH group, indicating that the traditional Chinese and Tibetan medicine compound preparation can enhance the activity of superoxide dismutase in female yaks and improve the ability to scavenge free radicals; the total antioxidant capacity (T-AOC) of the DE group was also significantly higher than that of the DH group, indicating that the traditional Chinese and Tibetan medicine compound preparation effectively improved the overall antioxidant level of female yaks; the glutathione peroxidase (GSH-Px) activity of the DE group increased significantly, further confirming the positive effect of the traditional Chinese and Tibetan medicine compound preparation on the antioxidant system of female yaks; while the content of malondialdehyde (MDA) in the DE group was significantly lower than that of the DH group, indicating that the traditional Chinese and Tibetan medicine compound preparation can reduce the generation of lipid peroxidation products and reduce oxidative stress. Through the above analysis, it can be known that by supplementing the traditional Chinese and Tibetan medicine compound preparation, the antioxidant ability of postpartum female yaks can be significantly improved. (ii) On the 15th and 30th days of the experiment, the anti-inflammatory related index interleukin-10 (IL-10) in the DE group increased significantly, while the inflammatory related factors interleukin-6 (IL-6) and interleukin-1β (IL-1β) decreased significantly; specifically, the IL-10 level in the DE group was significantly higher than that in the DH group, indicating that the traditional Chinese (Tibetan) medicine compound can promote the production of anti-inflammatory factors and help inhibit the inflammatory response; at the same time, the IL-6 and IL-1β levels in the DE group were significantly lower than those in the DH group, indicating that the traditional Chinese and Tibetan medicine compound preparation can effectively reduce the expression of inflammatory factors, thereby reducing the inflammation occurrence in postpartum weaned female yaks. It can be seen from this that supplementing the traditional Chinese and Tibetan medicine compound preparation prepared by the present invention can not only significantly enhance the antioxidant ability of postpartum female yaks, but also effectively reduce the inflammatory response, providing a positive protective effect on the health of female yaks.

[0055] (2) Figure 2 For the alpha diversity analysis of the rumen microorganisms of postpartum female yaks by the traditional Chinese and Tibetan medicine compound preparation on the 15th and 30th days of the experiment, it can be seen from the figure that: The species accumulation curve ( Figure 2 C) used to measure and predict the change of species richness with the increase of sample size shows that as the sample number increases, the curve tends to be flat, and the Good's coverage index of each group is close to 100%, indicating that the current sequencing depth is sufficient to cover all existing species, ensuring the reliability of the data. Figure 2In A, the α-diversity indices of the DE group and the DH group were compared. Specifically, the Chao1 index of the DE group increased significantly, which means that there are more operational taxonomic units (OTUs) in the rumen microorganisms, indicating a higher species richness; the Pielou evenness index of the DE group also increased significantly, indicating that the community composition is more uniform and the distribution of each species in the community is more balanced; although the Shannon index and Simpson index showed differences between the two groups, they did not reach a significant level, indicating that the DE group and the DH group have similar trends in species diversity and dominance; although the DE group has a larger number of OTUs and a more uniform community composition, compared with the DH group, this difference is not significant, indicating that the rumen microorganisms between the two groups show certain similarities in overall richness, diversity and evenness. Figure 2 B further shows the change trend of the Good's coverage index. The results show that the coverage of both groups is close to 100%, verifying the sufficiency of the sequencing depth again.

[0056] (3) As Figure 3 shown, the number of ASVs (amplicon sequence variants) in the DE1 and DE2 groups is higher than that in the DH1 and DH2 groups. Specifically, in Figure 3 A, the DE1 group has 11,554 unique ASVs, the DH1 group has 11,072 unique ASVs, and there are 3,868 ASVs in common between DE1 and DH1; in Figure 3 B, the DE2 group has 12,368 unique ASVs, the DH2 group has 10,897 unique ASVs, and there are 4,106 ASVs in common between DE2 and DH2. By comparing the number of ASVs and their common parts in the DE1, DE2 and DH1, DH2 groups, it can be seen that after adding the traditional Chinese and Tibetan medicine compound preparation, there are significant differences in the rumen microbial composition of the DE group. The DE group not only has a higher total number of ASVs than the DH group, but also shows higher diversity in the number of unique ASVs. Although there are some common ASVs between the two groups, the number of unique ASVs in the DE group is significantly more, indicating that the traditional Chinese and Tibetan medicine compound preparation prepared by the present invention has a positive promoting effect on the richness and diversity of rumen microorganisms.

[0057] (4) As Figure 4 shown, on the 15th day (DE1, DH1) and the 30th day (DE2, DH2) of the experiment, the rumen microbial composition structure of postpartum female yaks was analyzed by PCoA (principal coordinates analysis) and NMDS (non-metric multidimensional scaling analysis). In Figure 4In group A: On the 15th day of the experiment (DE1 and DH1), the points between the two groups were significantly separated in the PCoA space, indicating that there were significant differences in the rumen microbial composition structure between the DE1 group and the DH1 group after feeding the traditional Tibetan and Chinese compound preparation; on the 30th day of the experiment (DE2 and DH2), the points of the DE2 group and the DH2 group also showed obvious separation, further confirming the long-term effect of the traditional Tibetan and Chinese compound preparation on the rumen microbial composition. In summary, it can be known that for the postpartum female yaks fed with the traditional Tibetan and Chinese compound preparation, the composition structure of their rumen microorganisms changed significantly on the 15th and 30th days of the experiment. Both PCoA and NMDS analyses showed that the points of the DE group and the DH group were significantly separated, and the stress value of the NMDS analysis was lower than 0.2, further verifying the reliability of the analysis results; thus, it shows that the traditional Tibetan and Chinese compound preparation prepared by the present invention has a significant effect on the composition structure of the rumen microorganisms of postpartum female yaks.

[0058] (5) As Figure 5 shown, on the 15th day (DE1, DH1) and the 30th day (DE2, DH2), the top three compositions of the relative abundances of rumen microorganisms at the genus level in each group were roughly the same, but there were certain differences in their relative abundances. Specifically: (i) In the Figure 5 overall analysis of A, on the 15th day of the experiment, the dominant genera in both the DE1 group and the DH1 group were Limimorpha, SFMI01, Prevotella, and Cryptobacteroides. These dominant genera had relatively high relative abundances in both groups, indicating that they were the main components of rumen microorganisms during this period; on the 30th day of the experiment, the dominant genera in both the DE2 group and the DH2 group were still Limimorpha, SFMI01, Prevotella, and Cryptobacteroides, which were basically the same as the composition on the 15th day of the experiment; however, their relative abundances changed slightly. The relative abundances of Prevotella and SFMI01 in the DE group increased significantly, indicating that adding the traditional Tibetan and Chinese compound preparation prepared by the present invention had an obvious promoting effect on these genera. (ii) In the Figure 5 individual analysis of B, on the 30th day of the experiment, from each individual, the relative abundances of Limimorpha and Prevotella in the DE group also increased significantly, further confirming the significant effect of the traditional Tibetan and Chinese compound preparation prepared by the present invention on the rumen microbial composition.

[0059] (6) As Figure 6As shown in the figure, through LEfSe (Linear Discriminant Analysis Effect Size) analysis, it was found that: only the DH group had significant biomarkers, while the DE group did not have significant biomarkers. Specifically, the biomarkers in the DH1 group were o_LD1_PB3, g_Lenti_01, f_Lenti_01, and c_Kiritimatiellae_777785, and the relative abundances of these microorganisms in the DH1 group were significantly higher than those in other groups; the biomarkers in the DH2 group were f_Carnobacteriaceae, g_Carnobacterium_A_320743, o_Lactobacillales, p_Firmicutes_D, and c_Bacilli, and the relative abundances of these microorganisms in the DH2 group were significantly higher than those in other groups. In summary, it can be seen that adding the traditional Chinese and Tibetan medicine compound preparation prepared by the present invention has a significant impact on the rumen microbial composition of postpartum female yaks, especially the promoting effect on certain specific genera and taxonomic units.

[0060] Example 3

[0061] This example is about the effect of the traditional Chinese and Tibetan medicine compound preparation prepared in Example 1 above on the intestinal flora of postpartum female yaks.

[0062] Twenty postpartum female yaks weaned 80 ± 15 days after giving birth were selected for the experiment and randomly divided into a control group (DH) and a traditional Chinese and Tibetan medicine compound preparation supplementary feeding group (DE), with 10 yaks in each group. The traditional Chinese and Tibetan medicine compound preparation supplementary feeding group (DE) was fed a basal diet (5 kg) supplemented with 5% of the traditional Chinese and Tibetan medicine compound preparation prepared in Example 1 every day, while the blank control group (DH group) was fed 5 kg of basal diet every day. The environmental conditions were kept consistent in all groups. The first 7 days before the experiment were the adaptation period. From the 1st to the 4th day, 1 / 4 of the test dose of the traditional Chinese and Tibetan medicine compound preparation was given, and from the 5th to the 7th day, 1 / 2 of the test dose was given to observe whether there were any adverse reactions in the postpartum female yaks; starting from the 8th day, the formal experimental period began, and the dosing dose was adjusted to 5% of the traditional Chinese and Tibetan medicine compound preparation, and the experiment lasted for 30 days; the daily feeding time was 9:30 and 15:00 to ensure that the traditional Chinese and Tibetan medicine compound preparation was fully mixed with the basal diet, and the female yaks drank water freely during the experiment. On the 15th and 30th days of the experiment, 200 g of fresh rectal fecal samples of postpartum female yaks in each group were collected respectively; the samples collected on the 15th day were classified as DE1 (divided into DA1 - DA6 between groups) and DH1 (divided into DD1 - DD6 between groups), and the samples collected on the 30th day were classified as DE2 (divided into DA7 - DA12 between groups) and DH2 (divided into DD7 - DD12 between groups); these collected fresh fecal samples will be used for 16S rRNA high-throughput sequencing to deeply analyze the effect of the traditional Chinese and Tibetan medicine compound preparation prepared by the present invention on the intestinal flora of postpartum female yaks, and the specific analysis is as follows.

[0063] (1) AsFigure 7 As shown, the intestinal flora diversity and species richness of the DE group and the DH group were evaluated by multiple indices on the 15th and 30th days of the experiment. From the species accumulation curve ( Figure 7 C), it can be seen that as the sample size increases, the number of species gradually tends to be stable, indicating that the current sequencing depth is sufficient to cover all species; moreover, the Good's coverage index of each group is close to 100%, further verifying the sufficiency of the sequencing depth and ensuring the reliability of the data. In Figure 7 the comparison of α-diversity indices, on the 15th and 30th days of the experiment, there were no significant differences in the Simpson index, Shannon index, Pielou_e index, Faith_pd index, and Goods_coverage index between the DE group and the DH group (p > 0.05), indicating that the diversity levels of the two groups were similar in these aspects; however, there were significant differences in the Chao1 index (p = 0.017) and Observed_species index (p = 0.02) between the DE group and the DH group. Specifically, the Chao1 index of the DE group was significantly higher than that of the DH group, indicating that the DE group had a higher potential species richness; the Observed_species index of the DE group was also significantly higher than that of the DH group, further confirming its higher species richness. Through comprehensive analysis, although there were no significant differences in multiple diversity indices between the DE group and the DH group, the significant advantages of the DE group in the Chao1 index and Observed_species index indicated its higher species richness; thus, it shows that the Tibetan and Chinese medicine compound preparation prepared by the present invention has a positive effect on the intestinal flora richness of postpartum female yaks, which helps to improve their intestinal health and overall physiological state.

[0064] (2) As Figure 8 shown, it can be seen from the figure that: in Figure 8 A, the DE1 group had 6230 unique ASVs, the DH1 group had 6117 unique ASVs, and there were 2657 ASVs in common between the DE1 and DH1 groups; in Figure 8 B, the DE2 group had 5419 unique ASVs, the DH2 group had 5649 unique ASVs, and there were 2432 ASVs in common between the DE2 group and the DH2 group. In summary, the Tibetan and Chinese medicine compound preparation supplementary feeding group (DE group) and the blank control group (DH group) prepared by the present invention showed different intestinal flora composition characteristics at different stages of the experiment. Although there was an overlap in some common ASVs between the two groups, the DE group was richer in the number of unique ASVs; thus, it shows that the Tibetan and Chinese medicine compound preparation prepared by the present invention may have a positive effect on the intestinal flora diversity and richness of postpartum female yaks, which helps to improve their intestinal health status.

[0065] (3) As Figure 9As shown, on the 15th day (DE1, DH1) and the 30th day (DE2, DH2) of the experiment, the intestinal flora structure of postpartum female yaks was comparatively analyzed by PCoA (Principal Coordinates Analysis) and NMDS (Non-metric Multidimensional Scaling). At Figure 9 In A, in the PCoA graph, the points of the DE1 group and the DH1 group were significantly separated, indicating that there were significant differences in the intestinal flora structure between the two groups on the 15th day of the experiment, suggesting that the Tibetan and Chinese medicine compound preparation prepared by the present invention had begun to affect the composition and diversity of intestinal microorganisms; on the 30th day of the experiment (DE2 and DH2), the points of the DE2 group and the DH2 group also showed varying degrees of separation, further confirming that with the extension of the experimental time, the influence of the Tibetan and Chinese medicine compound preparation prepared by the present invention on the intestinal flora gradually emerged and persisted. At Figure 9 In the B NMDS analysis, the points of the DE1 group and the DH1 group still maintained an obvious separation trend, which was consistent with the results of the PCoA analysis, verifying again the significant differences in the intestinal flora structure between the two groups; the points of the DE2 group and the DH2 group also showed a certain degree of separation, further illustrating the long-term influence of the Tibetan and Chinese medicine compound preparation prepared by the present invention on the intestinal flora structure.

[0066] (4) As Figure 10 shown, the top two compositions of the relative abundances of the intestinal flora of each group at the genus level on the 15th day and the 30th day were roughly the same, but there were certain differences in their relative abundances. Specifically, (i) in the Figure 10 overall analysis of A, on the 15th day of the experiment, Faecousia, Cryptobacteroides, Arthrobacter_B, and Akkermansia were the four most common genera in the DE1 group and the DH1 group; among them, the relative abundance of Faecousia in the DE1 group increased significantly, indicating that the Tibetan and Chinese medicine compound preparation prepared by the present invention might have a promoting effect on Faecousia, increasing its proportion in the intestinal flora. On the 30th day of the experiment, these four dominant genera still existed in the DE2 group and the DH2 group, but their relative abundances changed slightly; the Faecousia in the DE2 group continued to maintain a high abundance, further verifying the continuous influence of the Tibetan and Chinese medicine compound preparation prepared by the present invention on it. (ii) In the Figure 10In the individual analysis of Group B, on the 15th day of the experiment, from the perspective of each individual, the relative abundance of Faecousia in the DE group generally increased, indicating that the traditional Tibetan and Chinese medicine compound preparation prepared by the present invention had a positive effect on Faecousia in different individuals; on the 30th day of the experiment, in addition to Faecousia, the relative abundance of Arthrobacter_B in the DE group also increased significantly, indicating that as the experiment time extended, the traditional Tibetan and Chinese medicine compound preparation prepared by the present invention not only continued to promote the growth of Faecousia, but also had a positive effect on Arthrobacter_B. This shows that the traditional Tibetan and Chinese medicine compound preparation prepared by the present invention can have a positive impact on the intestinal health of postpartum female yaks by regulating the abundance of specific genera of bacteria.

[0067] (5) As Figure 11 shown, through LEfSe (Linear Discriminant Analysis Effect Size) analysis, it was found that there were significant differences in the composition and structure of the intestinal flora between the DE group and the DH group, specifically manifested in the enrichment at different phylum and genus levels. In the DE1 group, bacteria such as c_Clostridia_258483 and p_Firmicute_A were significantly enriched, indicating that on the 15th day of the experiment, the relative abundance of these specific bacteria in the DE group was significantly higher than that in other groups; on the 30th day of the experiment, significant enrichment of bacteria such as g_Peribacillus301373, f_DSM_1321_301386, and o_Bacillales_B_302584 was detected in the DE2 group, indicating that as the experiment time extended, the impact of the traditional Tibetan and Chinese medicine compound preparation prepared by the present invention on the intestinal flora gradually emerged and persisted, and its promoting effect on specific bacteria. For the DH group: in the DH1 group, bacteria such as c_Bacilli and p_Firmicute_D were significantly enriched, indicating that on the 15th day of the experiment, the relative abundance of these bacteria in the DH group was relatively high; on the 30th day of the experiment, significant enrichment of bacteria such as f_Micrococcaceae and o_Actinomycetia was detected in the DH2 group, indicating that as time passed, the intestinal flora structure of the control group also changed, but this change trend was different from that of the DE group, reflecting the natural evolution process of the intestinal flora in the control group. This shows that the traditional Tibetan and Chinese medicine compound preparation prepared by the present invention significantly affected the intestinal flora structure of the DE group, promoted the enrichment of specific bacteria (such as c_Clostridia_258483, g_Peribacillus301373, etc.), and had specificity in regulating the intestinal flora of postpartum female yaks.

[0068] Example 4

[0069] In this example, the traditional Tibetan and Chinese medicine compound preparation prepared in Example 1 above was added to the basic feed for postpartum female yaks at a ratio of 5% for the experiment.

[0070] Twenty postpartum weaned female yaks 80 ± 15 days after parturition were selected for the experiment and randomly divided into a control group (DH) and a supplementary feeding group of the traditional Chinese and Tibetan medicine compound preparation (DE), with 10 heads in each group. The supplementary feeding group of the traditional Chinese and Tibetan medicine compound preparation (DE) was fed a basal diet (5 kg) supplemented with 5% of the traditional Chinese and Tibetan medicine compound preparation prepared in Example 1 every day, while the blank control group (DH group) was fed 5 kg of basal diet every day. The environmental conditions were kept consistent in all groups. The first 7 days before the start of the experiment were the adaptation period. On the 1st to 4th days, 1 / 4 of the experimental dose of the traditional Chinese and Tibetan medicine compound preparation was given, and on the 5th to 7th days, 1 / 2 of the experimental dose was given to observe whether there were any adverse reactions in the postpartum female yaks. From the 8th day, the experiment period officially began, and the dosing dose was adjusted to 5% of the traditional Chinese and Tibetan medicine compound preparation. The experiment lasted for 30 days, and the whole experimental cycle was 90 days. The daily feeding time was 9:30 am and 15:00 pm to ensure that the traditional Chinese medicine compound was fully mixed with the basal diet and to ensure that the female yaks could drink water freely during the experiment. On the 15th, 30th, 45th, and 60th days of the experiment, blood samples of all female yaks were collected. The blood collection method was jugular vein blood collection. After collection, the blood was allowed to stand at room temperature for 2 - 3 h, and then the serum was separated by centrifugation at a speed of 3500 r / min for 15 min for detecting the levels of estrogen (E2), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) to evaluate the effect of the traditional Chinese and Tibetan medicine compound preparation prepared by the present invention on the endocrine function of postpartum female yaks.

[0071] As Figure 12As shown, it can be seen from the figure that the levels of estrogen (E2), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) in the serum of female yaks in the DE group are significantly higher than those in the DH group, especially on the 45th and 60th days. Specifically, it is manifested as follows: (i) Estrogen (E2): On the 15th day, there is no obvious difference in the E2 level between the DE group and the DH group; on the 30th day, the E2 level in the DE group is slightly higher than that in the DH group; on the 45th day, the E2 level in the DE group is significantly higher than that in the DH group (p < 0.01), showing an obvious improvement effect; on the 60th day, the E2 level in the DE group continues to maintain at a high level and still has a significant difference compared with the DH group (p < 0.01). (ii) Luteinizing hormone (LH): On the 15th day, there is no significant difference in the LH level between the two groups; on the 30th day, the LH level in the DE group begins to be higher than that in the DH group; on the 45th day, the LH level in the DE group is significantly higher than that in the DH group (p < 0.05), indicating that the traditional Chinese and Tibetan medicine compound preparation prepared in the present invention has a promoting effect on the LH level; on the 60th day, the LH level in the DE group further increases and maintains a significant difference with the DH group (p < 0.05). (iii) Follicle-stimulating hormone (FSH): On the 15th day, there is no significant difference in the FSH level between the DE group and the DH group; on the 30th day, the FSH level in the DE group is slightly higher than that in the DH group; on the 45th day, the FSH level in the DE group is significantly higher than that in the DH group (p < 0.05), showing the promoting effect of the traditional Chinese and Tibetan medicine compound preparation prepared in the present invention on the FSH level; on the 60th day, the FSH level in the DE group continues to maintain at a high level and still has a significant difference compared with the DH group (p < 0.05).

[0072] Through comprehensive analysis, it can be known that after feeding the traditional Chinese and Tibetan medicine compound preparation prepared in the present invention, the levels of estrogen (E2), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) in the serum of female yaks are significantly higher than those of the control group of female yaks, especially on the 45th and 60th days of the experiment. This shows that the traditional Chinese and Tibetan medicine compound preparation prepared in the present invention can effectively regulate the endocrine function of postpartum female yaks, has a significant promoting effect on the reproductive hormone level, and provides a scientific basis for the health management and improvement of reproductive performance of postpartum female yaks.

[0073] Example 5

[0074] In this example, the traditional Chinese and Tibetan medicine compound preparation prepared in Example 1 above was added to the basic feed of postpartum female yaks at a ratio of 5% for testing.

[0075] Twenty postpartum weaned female yaks 80 ± 15 days after parturition were selected for the experiment and randomly divided into a control group (DH) and a supplementary feeding group of the traditional Chinese and Tibetan medicine compound preparation (DE), with 10 yaks in each group. The supplementary feeding group of the traditional Chinese and Tibetan medicine compound preparation (DE) was fed a basal diet (5 kg) supplemented with 5% of the traditional Chinese and Tibetan medicine compound preparation prepared in Example 1 every day, while the blank control group (DH group) was fed 5 kg of the basal diet every day. The environmental conditions were kept consistent in all groups. The first 7 days before the start of the experiment were the adaptation period. On the 1st - 4th days, 1 / 4 of the experimental dose of the traditional Chinese and Tibetan medicine compound preparation was given, and on the 5th - 7th days, 1 / 2 of the experimental dose was given to observe whether there were any adverse reactions in the postpartum female yaks. From the 8th day, the experiment period officially began, and the dosing dose was adjusted to 5% of the traditional Chinese and Tibetan medicine compound preparation. The experiment lasted for 30 days, and the whole experimental cycle was 90 days. The daily feeding time was 9:30 am and 15:00 pm to ensure that the traditional Chinese medicine compound was fully mixed with the basal diet and to ensure that the female yaks could drink water freely during the experiment. During the experiment, the time of the female yaks' estrus recurrence and behavioral characteristics were closely observed and recorded, including abnormal behaviors such as restlessness, jumping, running, etc., sniffing, biting or licking companions, actively approaching or chasing bulls; accepting or actively performing mounting behaviors; congestion, swelling of the vulva accompanied by the discharge of transparent or viscous secretions; and making low growls or long howls and other behaviors different from the daily ones for statistical analysis.

[0076] As Figure 13 shown, it can be seen from the figure that the average estrus time of the DE group (supplementary feeding group of the traditional Chinese and Tibetan medicine compound preparation) was about 60 days, and the average estrus time of the DH group (blank control group) was about 68 days. This indicates that the estrus time of the DE group was significantly shorter than that of the DH group. The DE group came into estrus about 8 days earlier on average than the DH group, and the difference was statistically significant (p < 0.01). In summary, it can be known that after feeding the traditional Chinese and Tibetan medicine compound preparation prepared by the present invention, it can effectively promote the estrus cycle of postpartum female yaks and shorten the time of postpartum female yaks' estrus recurrence (p < 0.01). This result shows that the traditional Chinese and Tibetan medicine compound preparation prepared by the present invention has a significant effect on promoting the reproductive health of postpartum female yaks, providing a scientific basis for improving the reproductive efficiency of yaks.

[0077] The above has introduced in detail a traditional Chinese and Tibetan medicine compound preparation for promoting postpartum recovery of yaks and shortening the estrus cycle, its preparation method and application. Specific examples are used in this article to elaborate on the principle and implementation mode of the present invention. The description of the above examples is only used to help understand the method and its core idea of the present invention. It should be noted that for those of ordinary skill in the art in this technical field, without departing from the principle of the present invention, several improvements and modifications can still be made to the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.

Claims

1. A Chinese and Tibetan medicine compound preparation for promoting postpartum recovery of yaks and shortening the estrus cycle, characterized in that: The Chinese-Tibetan medicine compound preparation is prepared from the following raw materials in parts by weight: 15-20 parts of scutellaria baicalensis, 15-20 parts of codonopsis pilosula, 15-20 parts of astragalus, 15-20 parts of white peony root, 5-10 parts of polygonatum sibiricum and 5-10 parts of liquorice.

2. The Chinese-Tibetan medicinal compound preparation for promoting postpartum recovery of yaks and shortening the estrus cycle according to claim 2, characterized in that: The Chinese and Tibetan medicine compound preparation is prepared from the following raw materials in parts by weight: 18 parts of scutellaria baicalensis, 18 parts of codonopsis pilosula, 18 parts of astragalus, 18 parts of white peony root, 7 parts of polygonatum sibiricum and 7 parts of liquorice.

3. The method for preparing the Chinese-Tibetan medicinal compound preparation for promoting postpartum recovery of yaks and shortening the estrus cycle according to any one of claims 1 to 2, characterized in that: The following steps are involved: S1. Weigh appropriate amounts of Scutellaria baicalensis, Codonopsis pilosula, Astragalus membranaceus, Paeonia lactiflora, Polygonatum sibiricum, and Licorice by weight, dry each herb one by one, and then crush the dried herbs to obtain herb powder; S2. The powders of the medicinal materials crushed in step S1 are screened through a sieve respectively, and then each of the screened medicinal material powders is evenly mixed according to the above weight proportions to obtain a Chinese Tibetan medicine composite preparation.

4. The method for preparing the Chinese-Tibetan medicinal compound preparation for promoting postpartum recovery of yaks and shortening the estrus cycle according to claim 3, characterized in that: In step S2, the screened medicinal material powders are mixed in parts by weight, including 15 to 20 parts of Scutellaria baicalensis, 15 to 20 parts of Codonopsis pilosula, 15 to 20 parts of Astragalus membranaceus, 15 to 20 parts of Paeonia lactiflora, 5 to 10 parts of Polygonatum sibiricum, and 5 to 10 parts of Licorice.

5. The method for preparing the Chinese-Tibetan medicinal compound preparation for promoting postpartum recovery of yaks and shortening the estrus cycle according to claim 3, characterized in that: In step S2, the mesh size of the sieve is 30 to 50 meshes.

6. Use of the Chinese-Tibetan medicinal compound preparation according to any one of claims 1 to 2 in the preparation of a drug for promoting postpartum recovery or shortening the estrus cycle of yaks.

7. The use according to claim 6, characterized in that: The Chinese and Tibetan medicine compound preparation is used in preparing medicines or feed additives for postpartum weaned yaks.

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