Green compound micro-ecological preparation for improving growth performance and intestinal flora of yaks and application of green compound micro-ecological preparation

Abstract

The invention provides a green composite microecological preparation for improving the growth performance and intestinal flora of yaks and application of the green composite microecological preparation, and belongs to the technical field of livestock breeding. The green composite microecological preparation provided by the invention comprises the following raw materials in parts by mass: 20-25 parts of corn, 8-10 parts of corn germ meal, 4-7 parts of wheat middling, 6-8 parts of soybean meal and 1-3 parts of cottonseed protein. The green composite microecological preparation is prepared by adding the raw materials into a mixed bacteria solution and fermenting. The green compound micro-ecological preparation provided by the invention can endow feed with a micro-ecological regulation function, promote digestion and absorption of nutrients, improve the feed conversion rate, promote growth and development of yaks, shorten the breeding cycle and improve economic benefits.

Description

Technical Field

[0001] The present invention relates to the technical field of livestock breeding, and particularly to a green compound microecological preparation for improving the growth performance and intestinal flora of yaks and its application. Background Art

[0002] During the process of yak breeding, the existing ordinary feed cannot fully meet the nutritional requirements for the growth and development of yaks, resulting in a slow growth rate of yaks and a low feed conversion efficiency. At the same time, in terms of maintaining the health of the yak body and optimizing the rumen function, ordinary feed cannot effectively regulate the rumen microflora and metabolism, affecting the digestion and absorption of feed by yaks. Therefore, it is usually necessary to add concentrated supplements for yaks to further supplement nutrition. However, traditional concentrated supplements for yaks have certain limitations in terms of nutrient utilization efficiency and promoting the growth of yaks, restricting the improvement of yak growth performance and increasing breeding costs. Moreover, they have poor effects in maintaining the health of the yak body and improving the rumen function, and cannot meet the requirements of modern yak breeding for efficient and healthy breeding. Therefore, providing a feed additive that can comprehensively improve the production performance of yaks and optimize the rumen environment is of great significance for improving the breeding efficiency of yaks. Summary of the Invention

[0003] The purpose of the present invention is to provide a green compound microecological preparation for improving the growth performance and intestinal flora of yaks, improving the feed conversion efficiency, promoting the growth and development of yaks, and enhancing the physique and stress resistance of yaks.

[0004] In order to achieve the above-mentioned invention purpose, the present invention provides the following technical solutions:

[0005] The present invention provides a green compound microecological preparation for improving the growth performance and intestinal flora of yaks, which is characterized by comprising the following raw materials in parts by mass: 20 - 25 parts of corn, 8 - 10 parts of corn germ meal, 4 - 7 parts of wheat middlings, 6 - 8 parts of soybean meal, and 1 - 3 parts of cottonseed protein;

[0006] After adding the above raw materials to the mixed bacterial liquid and fermenting, it becomes the green compound microecological preparation.

[0007] Preferably, the mixed bacterial liquid is composed of Bacillus licheniformis, Candida utilis, and Saccharomyces cerevisiae in a volume ratio of 2 - 3:1 - 2:1 - 2.

[0008] Preferably, the bacterial concentrations of Bacillus licheniformis, Candida utilis, and Saccharomyces cerevisiae in the mixed bacterial liquid are all ≥ 1.0×10 7 CFU / mL.

[0009] Preferably, the inoculation amount of the mixed bacterial liquid is 1% - 3% v / w of the total weight of the raw materials, and the fermentation conditions are as follows: the moisture content is 55% - 60%, the fermentation temperature is 25 - 30 °C, and the fermentation time is 20 - 30 d.

[0010] The present invention also provides the application of the described green composite microecological preparation in preparing a feed for improving the growth performance and intestinal flora of yaks. The feed includes roughage, concentrate, and the green composite microecological preparation.

[0011] Preferably, the mass ratio of the roughage, concentrate, and green composite microecological preparation is 45 - 50:25 - 30:20 - 25.

[0012] Preferably, the roughage is wheat hay, and the concentrate includes 20 - 25 parts of corn, 8 - 10 parts of corn germ meal, 4 - 7 parts of wheat middlings, 6 - 8 parts of soybean meal, 1 - 3 parts of cottonseed protein, 0.3 - 1 part of salt, 0.1 - 0.2 part of calcium hydrogen phosphate, and 1 - 3 parts of premix.

[0013] The green composite microecological preparation for improving the growth performance and intestinal flora of yaks provided by the present invention endows the feed with a microecological regulation function through the fermentation of the mixed bacterial liquid, improves the digestion and absorption efficiency of animals for feed nutrients, enhances the feed conversion efficiency, promotes the growth and development of yaks, shortens the breeding cycle, and improves the breeding economic benefits. It can further improve the nutritional value of the feed, enhance the physical fitness of animals, improve the stress resistance, comprehensively improve the animal health level and reproductive performance, and promote the green, efficient, and sustainable development of the livestock breeding industry. And the use of microbial fermentation technology to prepare feed avoids the potential hazards brought by the use of chemical synthetic additives, conforms to the concept of green breeding, and ensures the quality and safety of livestock products. Description of the Drawings

[0014] Figure 1 It is the veen diagram of the rumen microbial flora composition in Example 2;

[0015] Figure 2 It is the diagram of the rumen microbial flora composition at the phylum level in Example 2;

[0016] Figure 3 It is the diagram of the rumen microbial flora composition at the genus level in Example 2;

[0017] Figure 4 It is the analysis diagram of the difference in the abundance of rumen microorganisms at the genus level in Example 2;

[0018] Figure 5 It is the LEfSe analysis diagram of rumen microorganisms from the phylum level to the genus level in Example 2. Detailed Embodiments

[0019] The technical solution provided by the present invention will be described in detail below in conjunction with embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0020] Example 1

[0021] 1.1 Strain activation

[0022] Use an inoculation loop to separately pick up Bacillus licheniformis, Candida utilis, and Saccharomyces cerevisiae preserved on the plate, inoculate them into the corresponding culture media, and activate and culture for 24 h to obtain the primary seed liquid. Pipette 1% of the primary seed liquid into the corresponding liquid culture medium and culture for 30 h to make the bacterial concentration reach 1×10 7 CFU / mL as the secondary seed liquid. Inoculate the secondary seed liquid into the corresponding culture medium, perform enlarged culture, and then mix Candida utilis, Saccharomyces cerevisiae, and Bacillus licheniformis in a volume ratio of 2:1:1 to obtain a mixed bacterial liquid (the bacterial concentrations of all three bacteria are ≥1.0×10 7 CFU / mL). The culture media and culture conditions used in the above culture process are shown in Table 1.

[0023] Table 1 Culture media and culture conditions corresponding to the three bacteria

[0024] Strain category Culture medium Cultivation conditions Bacillus licheniformis TSA 37 °C, 160 r / min Candida utilis NB / YM 30 °C, 160 r / min Saccharomyces cerevisiae Malted wort 30 °C, 160 r / min

[0025] 1.2 Feed fermentation

[0026] Pour corn, corn germ meal, wheat middlings, soybean meal, and cottonseed protein into the stirring tank in sequence, stir, and after stirring evenly, add the above-mentioned mixed bacterial liquid, with the addition amount being 1% v / w of the total weight of the raw materials, and stir again to make them fully mixed. Pack the mixed material into a breathing bag and seal it for fermentation treatment. Control the moisture content at 55% - 60% and the temperature below 28°C during fermentation. Ferment for 30 d to obtain a green composite microecological preparation. Measure the changes in nutrients during fermentation. As shown in Table 2.

[0027] Table 2 Changes in nutrients on the 0th, 7th, and 30th days of fermentation

[0028] Item Fermentation day 0 Fermentation day 7 Fermentation day 30 Ash content <![CDATA[12.58±0.5 a > <![CDATA[7.35±0.09 b > <![CDATA[6.97±0.05 b > Protein <![CDATA[18.77±0.1 b > <![CDATA[19.65±0.36 b > <![CDATA[25.15±2.15 a <!-- 2 -->]]> Crude fat <![CDATA[4.98±0.70 a > <![CDATA[4.81±0.50 a > <![CDATA[5.44±0.54 a > Calcium <![CDATA[0.24±0.12 b > <![CDATA[0.46±0.13 a > <![CDATA[0.59±0.05 a > Phosphorus <![CDATA[0.92±0.16 a > <![CDATA[0.53±0.02 b > <![CDATA[0.44±0.00 b > Moisture content <![CDATA[54.95±0.89 a > <![CDATA[52.62±0.68 b > <![CDATA[48.35±0.66 c >

[0029] Note: Different letters indicate significant differences in content before and after fermentation.

[0030] 1.3 Optimization of fermentation bacterial liquid

[0031] Ferment using the different fermentation bacteria combinations of Comparative Example 1 (Bacillus licheniformis + Candida utilis), Comparative Example 2 (Bacillus licheniformis + Saccharomyces cerevisiae), Comparative Example 3 (Candida utilis + Saccharomyces cerevisiae), and Comparative Example 4 (Bacillus subtilis + Candida utilis + Saccharomyces cerevisiae) respectively in the manner of 1.2. After fermentation for 30 days, a green composite microecological preparation is obtained, and the nutrient content is measured, as shown in Table 3.

[0032] Table 3 Nutrient content of the green composite microecological preparation fermented with different fermentation bacteria combinations

[0033] Item Control example 1 Control example 2 Control example 3 Control example 4 Ash content <![CDATA[8.45±0.03 a > <![CDATA[9.53±0.05 a > <![CDATA[9.97±0.05 a > <![CDATA[7.34±0.04 b > Protein <![CDATA[19.54±1.84 a > <![CDATA[20.42±2.04 a > <![CDATA[20.15±1.95 a > <![CDATA[22.31±1.74 b > Crude fat <![CDATA[5.01±0.47 a > <![CDATA[5.03±0.45 a > <![CDATA[5.09±0.51 a > <![CDATA[5.12±0.51 a > Calcium <![CDATA[0.51±0.04 a > <![CDATA[0.47±0.03 a > <![CDATA[0.49±0.02 a > <![CDATA[0.53±0.02 a > Phosphorus <![CDATA[0.62±0.02 a > <![CDATA[0.72±0.01 b > <![CDATA[0.67±0.03 a > <![CDATA[0.54±0.02 b > Moisture content <![CDATA[49.24±0.61 a > <![CDATA[47.33±0.54 a > <![CDATA[48.21±0.64 a > <![CDATA[48.15±0.47 a >

[0034] Note: Different letters indicate significant differences between groups

[0035] By comparing Table 3 and Table 2, it can be seen that whether any strain is removed from the mixed bacterial liquid used for fermentation or the strain is replaced, the fermentation effect of the bacterial liquid is significantly reduced, and the nutrient content such as protein and crude fat in the green composite microecological preparation is significantly reduced. It can be seen that there is a synergistic effect between Bacillus licheniformis, Candida utilis, and Saccharomyces cerevisiae, and a green composite microecological preparation with rich nutrient content can be fermented and prepared only when the three strains are used together.

[0036] Example 2 Feeding experiment

[0037] 1.1 Feeding grouping

[0038] Select 30 healthy male yaks with similar ages and weights (weight 96.54 ± 13.52 kg), and randomly divide them into 5 groups, with 6 yaks in each group, namely the basal diet group (control group), the fermented feed group (experimental group), the Comparative Example 1 group, the Comparative Example 2 group, and the Comparative Example 3 group.

[0039] 1.2 Feeding management method

[0040] The control group was fed a basal diet (with the mass ratio of roughage to concentrate being 50:50), the treated group was fed fermented feed (replacing 25 parts of concentrate with a green compound microecological preparation, i.e., the mass ratio of roughage, concentrate, and green compound microecological preparation being 50:25:25), the comparative example 1 group was fed fermented feed (replacing 10 parts of concentrate with a green compound microecological preparation, i.e., the mass ratio of roughage, concentrate, and green compound microecological preparation being 50:40:10), the comparative example 2 group was fed fermented feed (replacing 20 parts of concentrate with a green compound microecological preparation, i.e., the mass ratio of roughage, concentrate, and green compound microecological preparation being 50:30:20), and the comparative example 3 group was fed fermented feed (replacing 40 parts of concentrate with a green compound microecological preparation, i.e., the mass ratio of roughage, concentrate, and green compound microecological preparation being 50:10:40). The experimental period included a 10-day pre-feeding period and a 90-day formal period. The experimental cattle were fed in separate pens and fed twice a day at 07:00 and 17:00. During the feeding period, clean and fresh drinking water was ensured, disinfection, ventilation, and cattle shed cleaning were well done to prevent disease transmission. Relevant indicators were measured at the end of the experiment. The formulas of roughage and concentrate are shown in Table 4 below.

[0041] Table 4 Formulas and Nutritional Components of Roughage and Concentrate

[0042]

[0043] Note: Each kilogram of premix contains 211.3 mg of Fe, 28.69 mg of Cu, 67.47 mg of Mn, 159.82 mg of Zn, 1.11 mg of I, 0.29 mg of Se, 0.84 mg of Co, 0.49 mg of Mo, V A 6580 IU, V D 540 IU and V E 73.11 mg (purchased from Yangling Tiandi Animal Nutrition Technology Co., Ltd.); the net energy was calculated with reference to NRC (2001).

[0044] 1.3 Growth Performance

[0045] On the 1st and 90th days of the feeding experiment, the empty stomach weights of the experimental cattle were measured before morning feeding, and the average daily gain (ADG), average daily dry matter intake (ADFI), and feed to gain ratio (F / G) of the cattle were calculated.

[0046] Average daily gain = (final weight - initial weight) / number of experimental days

[0047] Average daily dry matter intake = (total feed amount per group - total remaining feed amount) / (number of feeding days × number of fed heads)

[0048] Feed to weight ratio = Average daily feed intake / Average daily gain

[0049] The test results are shown in Table 5 below.

[0050] Table 5 Data analysis of growth performance

[0051]

[0052] Note: * indicates p < 0.05 compared with the control group.

[0053] As can be seen from Table 5, compared with the control group, the feed to weight ratio of the experimental group decreased significantly, and the average daily gain increased significantly (p < 0.05). After reducing the addition amount of the green compound microecological preparation in the comparative example 1 group, compared with the control group, the feed to weight ratio hardly changed, and the average daily gain increased slightly. It can be seen that the green compound microecological preparation has a certain effect on promoting growth, but the addition amount is too low and the effect is not obvious; after significantly increasing the addition amount of the green compound microecological preparation in the comparative example 3 group, compared with the control group, the feed to weight ratio increased significantly, while the average daily gain decreased significantly, that is, after excessive addition of the green compound microecological preparation, it is not conducive to the growth of yaks.

[0054] 1.4 Determination of rumen microbiota

[0055] Before the morning feeding on the 90th day of the experiment, the yaks were fixed with a neck clamp, and 100 mL of rumen fluid was collected from each experimental cattle using a gastric tube rumen fluid sampler. The collected rumen fluid was filtered through four layers of gauze and then aliquoted into cryopreservation tubes for storage. After thawing the rumen fluid in the cryopreservation tubes, 16S rRNA sequencing was performed, and the sequencing was completed by Shanghai Majorbio Bio-pharm Technology Co., Ltd. The relative abundances at the phylum level and genus level were analyzed. The results are as Figures 1 - 5 shown.

[0056] From Figure 1 it can be seen that the microbiota of the control group consisted of 6,192 ASVs, and the unique ASVs were 4,780. The microbiota of the experimental group consisted of 7,094 ASVs, and the unique ASVs were 5,682.

[0057] From Figure 2 it can be seen that the top four phyla with the highest abundances in the rumen microbiota of the two groups were Bacteroidota, Firmicutes, Patescibacteria, and Spirochaetota. Compared with the control group, the relative abundances of Firmicutes, Patescibacteria, and Spirochaetota in the experimental group were all significantly increased (P < 0.05).

[0058] It can be seen from Figure 3 that the top five genera with the highest abundances in the two groups of rumen microorganisms are Prevotella, Rikenellaceae_RC9_gut_group, Prevotellaceae-UCG-003, Prevotellaceae-UCG-001, and Christensenellaceae_R-7_group in sequence. Compared with the control group, the relative abundances of Rikenellaceae_RC9_gut_group and Christensenellaceae_R-7_group in the experimental group were both significantly increased (P<0.05).

[0059] It can be seen from Figure 4 that compared with the control group, the relative abundances of Butyrivibrio, Lachnospiraceae_FCS020_group, and Moryella in the experimental group were all significantly increased (P<0.05).

[0060] It can be seen from Figure 5 that the differential taxa in the control group include g__U29-B03, o__Veillonellales-Selenomonadales, f__Selenomonadaceae (P<0.05). The differential taxa in the experimental group include c__Bacilli, o__Clostridia_UCG-014, o__RF39, o__Erysipelotrichales, f__Erysipelatoclostridiaceae, g__UCG-004.

[0061] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A green composite microecological preparation for improving the growth performance and intestinal flora of yaks, characterized in that, It contains the following raw materials in parts by mass: 20 - 25 parts of corn, 8 - 10 parts of corn germ meal, 4 - 7 parts of wheat middlings, 6 - 8 parts of soybean meal, and 1 - 3 parts of cottonseed protein; The green composite microecological preparation is obtained by adding the above raw materials to a mixed bacterial solution for fermentation.

2. The green composite microecological preparation according to claim 1, wherein, The mixed bacterial solution is composed of Bacillus licheniformis, Candida utilis, and Saccharomyces cerevisiae in a volume ratio of 2 - 3:1 - 2:1 - 2.

3. The green composite microecological preparation according to claim 2, wherein The bacterial concentrations of Bacillus licheniformis, Candida utilis and Saccharomyces cerevisiae in the mixed bacterial solution are all ≥ 1.0×10 7 CFU / mL.

4. The green composite microecological preparation according to claim 3, wherein The inoculation amount of the mixed bacterial solution is 1% - 3% v / w of the total weight of the raw materials, and the fermentation conditions are as follows: the moisture content is 55% - 60%, the fermentation temperature is 25 - 30 °C, and the fermentation time is 20 - 30 d.

5. Use of the green composite microecological preparation according to any one of claims 1 to 4 in the preparation of a feed for improving the growth performance and intestinal flora of yaks, characterized in that The feed includes roughage, concentrate, and the green composite microecological preparation.

6. The application according to claim 5, wherein The mass ratio of the roughage, concentrate, and green composite microecological preparation is 45 - 50:25 - 30:20 - 25.

7. The application according to claim 6, characterized in that, The roughage is wheat hay, and the concentrate includes 20 - 25 parts of corn, 8 - 10 parts of corn germ meal, 4 - 7 parts of wheat middlings, 6 - 8 parts of soybean meal, 1 - 3 parts of cottonseed protein, 0.3 - 1 part of table salt, 0.1 - 0.2 part of calcium hydrogen phosphate, and 1 - 3 parts of premix.

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