A microecological composite preparation for improving boar semen quality and its preparation method and application

Abstract

The present invention discloses a microecological composite preparation for improving the quality of boar semen and its preparation method and application, which belongs to the field of biological agricultural microecological preparation and animal husbandry breeding technology, including: fermenting with probiotics and cyperus juncea meal to obtain cyperus juncea fermented product; mixing cyperus juncea fermented product with tryptone, liver extract, L-cysteine, and fructose to obtain the inner layer mixture of the microecological composite preparation; preparing a barrier film colloid, using the barrier film colloid to coat the surface of the inner layer mixture of the microecological composite preparation with a barrier film; coating the surface film on the outside of the barrier film to obtain the microecological composite preparation. After feeding the feed mixed with the microbial composite preparation of the present invention, the various indicators of the boar's sperm motility, sperm density, forward motility sperm count, and sperm deformity rate have been significantly improved, which significantly improves the semen quality of the boar.

Description

Technical Field

[0001] The present invention belongs to the field of biological agricultural microecological preparations and animal husbandry breeding technology, and more specifically, the present invention relates to a microecological composite preparation for improving the quality of boar semen, and a preparation method and application thereof. Background Art

[0002] Boar reproductive capacity is a key factor affecting the annual productivity of sows. Low sperm motility and high deformity rate are the main reasons for the reduction of boar reproductive capacity. Improving boar reproductive capacity through nutritional means is a research hotspot in animal nutrition regulation. As a potential feed resource with broad application prospects, Cyperus oleiferus has a role in improving male animal sexual desire, semen quality, sperm in vitro fertilization ability, removing reactive oxygen and lipid peroxidation, and preventing sperm oxidation loss. However, there are no reports on whether adding Cyperus oleiferus to the diet can improve boar reproductive capacity. Among them, Cyperus oleiferus ( Cyperus esculentus L. ), also known as tiger nut, is a perennial herbaceous plant of the Cyperus genus of the Cyperaceae family. Cyperus oleifera has the advantages of wide adaptability, short growth period, large biomass, and high oil content. Cyperus oleifera stems and leaves can be used as silage, and tubers can be eaten raw or cooked. In addition, Cyperus oleifera can be used to extract oil, and the oil cake after oil extraction can be used as feed. Therefore, Cyperus oleifera, as an emerging crop that integrates grain, oil, animal husbandry, and feed, has high economic value and development potential.

[0003] At the same time, to ensure that boars have good reproductive capacity and high semen quality, it is also necessary to ensure that boars have good growth status, which makes it particularly important to ensure the microecological balance in the boar's intestines. To keep the microbial community in the boar's intestines healthy and stable, adding a certain amount of microecological preparations during feeding is a common method. Probiotics in microecological preparations can produce hydrolases, fermentation enzymes and respiratory enzymes, which are beneficial to the degradation of protein, fat and more complex carbohydrates in feed. In addition, microecological preparations have no toxic side effects and have the advantages of regulating the balance of intestinal flora, promoting animal growth, improving feed conversion rate and improving the immune function of boars. Since the pig's gastric juice is highly acidic, its pH value is between 2 and 3, and the microecological preparations will stay in the boar's stomach for a long time, so after the outermost layer of biomolecular polymers is eroded by gastric acid, the microecological preparations inside will be eroded by gastric acid, resulting in the probiotics losing their activity before entering the intestines, and the effect of the microecological preparations will be greatly reduced. Therefore, it is necessary to add a barrier film to the microbial preparations to protect the probiotics to a certain extent. Meanwhile, there are no reports on the use of cyperus oleifera and microbial preparations together as feed additives for boars. Summary of the invention

[0004] An object of the present invention is to solve at least the above-mentioned problems and / or disadvantages and to provide advantages which will be described hereinafter.

[0005] In order to achieve these purposes and advantages according to the present invention, a method for preparing a microecological composite preparation for improving the quality of boar semen is provided, comprising the following steps:

[0006] Step 1: fermenting the cyperus oleifera meal with probiotics to obtain cyperus oleifera fermented product;

[0007] Step 2: mixing the fermented product of Cyperus oleifera with tryptone, liver extract, L-cysteine ​​and fructose to obtain an inner layer mixture of the microecological composite preparation;

[0008] Step 3, preparing a barrier film colloid, and using the barrier film colloid to coat the surface of the inner layer mixture of the microecological composite preparation with a barrier film;

[0009] Step 4: Coat the surface film on the outside of the barrier film to obtain a microecological composite preparation.

[0010] Preferably, the specific method of step 1 includes:

[0011] S11, selecting bacterial strains for activation respectively to obtain activated bacterial strains;

[0012] S12, crushing cyperus chinensis meal and sieving through 120 mesh to obtain cyperus chinensis powder; adding calcium carbonate and calcium hydrogen phosphate to the cyperus chinensis powder, then inoculating activated bacteria, stirring evenly, placing in a constant temperature incubator for culture at a temperature of 37° C., and fermenting for 7 to 16 days; wherein the mass ratio of calcium carbonate, calcium hydrogen phosphate and cyperus chinensis powder is 1 to 5:1 to 3:1000 to 2000.

[0013] Preferably, in S11, the bacterial species is a mixed bacteria of Lactobacillus plantarum and Lactobacillus salivarius, and the ratio of the inoculation volume of the activated bacterial solution of Lactobacillus plantarum and Lactobacillus salivarius is 1:2-4.

[0014] Preferably, in the step 2, the mass ratio of the cyperus oleifera fermentation product to tryptone, liver extract, L-cysteine ​​and fructose is 2-5:0.05-0.1:0.3-1:0.1-0.2:0.2-1.

[0015] Preferably, in step 3, the method for preparing the barrier film colloid comprises:

[0016] S31, weighing 2-4 parts of modified konjac glucomannan, 1-2 parts of starch, 1-2 parts of soybean flour, and 0.2-0.5 parts of chitosan by weight;

[0017] S32, first dispersing starch in 200-300 parts of deionized water to obtain a starch dispersion with a concentration of 2-10 mg / mL; heating the starch dispersion to 40-70° C., stirring continuously at a speed of 200-500 rpm, adding modified konjac glucomannan, and cooling to room temperature after keeping warm for 10-20 min, stopping stirring, and obtaining a gel system;

[0018] S33. Add soybean powder and chitosan to the gel system, disperse them ultrasonically at a frequency of 60-80 kHz for 1-3 hours, and then freeze-dry them at -20--4°C to obtain a barrier film colloid with a water content of 25%-33%.

[0019] Preferably, in said S31, the preparation method of modified konjac glucomannan comprises:

[0020] S311, using konjac glucomannan powder to prepare a konjac glucomannan solution with a concentration of 1.5-2wt%, adding sodium carbonate thereto to adjust the pH of the konjac glucomannan solution to 9-10, stirring for 2h, and leaving it to stand for 24-36h;

[0021] S312, adding guar gum to the konjac glucomannan solution to make the concentration of guar gum be 0.5-1wt%;

[0022] S313, adding nano silicon dioxide powder and sodium tripolyphosphate to the mixed solution, stirring and heating to 50-80 ℃ with a rotating speed of 600-1200 rpm, keeping warm for 2-5 hours, cooling to room temperature, evaporating and concentrating, and drying to obtain modified konjac glucomannan; wherein the mass ratio of konjac glucomannan powder, nano silicon dioxide powder and sodium tripolyphosphate is 4:0.5-1:0.05-0.1.

[0023] Preferably, in said S313, the particle size of the nano silicon dioxide powder is 20-80 nm, wherein the D90 particle size is below 55 nm.

[0024] Preferably, the preparation method of the surface film comprises: weighing 0.2-0.6 parts of xanthan gum, 0.2-0.4 parts of sodium carboxymethyl cellulose, 0.3-0.8 parts of jatropha meal, and 1-1.5 parts of starch by weight, adding them into 10-12 parts of sterile water, stirring evenly to obtain a crude liquid, evaporating the crude liquid and coating it on the surface of the barrier film to obtain the surface film.

[0025] A microecological composite preparation for improving boar semen quality is prepared by a method for preparing a microecological composite preparation for improving boar semen quality.

[0026] An application of a microecological composite preparation for improving the quality of boar semen, wherein the microecological composite preparation is applied as an additive to boar feed, and the mixing ratio of the microecological composite preparation to feed is 0.003-0.005:1; wherein the storage condition of the microecological composite preparation is dry storage at 2-4°C away from light, and the storage time does not exceed 7 days.

[0027] The present invention has at least the following beneficial effects:

[0028] The present invention uses cyperus oleifera meal as the main raw material of the microecological composite preparation, ferments the cyperus oleifera meal after mixed inoculation with Lactobacillus plantarum and Lactobacillus salivarius, mixes the fermented cyperus oleifera meal fermentation product with tryptone, liver extract, L-cysteine ​​and fructose to serve as the inner layer mixture of the microecological composite preparation; the results show that after feeding the feed mixed with the microbial composite preparation of the present invention, various indicators of the boar's sperm motility, sperm density, forward-moving sperm count and sperm deformity rate are significantly improved, and the semen quality of the boar is significantly improved.

[0029] At the same time, in order to ensure that the microecological composite preparation has gastric acid resistance and that the probiotics in the microecological composite preparation are not prematurely inactivated, the present invention uses a barrier film to coat the inner layer mixture of the microecological composite preparation; wherein the barrier film uses modified konjac glucomannan as a main raw material, utilizes the good film-forming property of konjac glucomannan and the property that it is not easily decomposed by digestive enzymes in gastric acid, uses nano-silicon dioxide powder and guar gum to cross-link and modify the konjac glucomannan, reduces the swelling property of konjac glucomannan, further improves the gastric acid resistance of the barrier film, ensures the survival number of Lactobacillus plantarum and Lactobacillus salivarius, enables most of Lactobacillus plantarum and Lactobacillus salivarius to reach the intestinal tract of the boar for fixed-point release and proliferation, so as to achieve the purpose of maintaining the microecological balance of the intestinal colony of the boar, and is conducive to making the boar reach a normal growth state, so as to produce high-quality semen.

[0030] Other advantages, objectives and features of the present invention will be embodied in part through the following description, and in part will be understood by those skilled in the art through study and practice of the present invention. DETAILED DESCRIPTION

[0031] The present invention is described in further detail below so that those skilled in the art can implement it according to the description.

[0032] It should be understood that the terms such as “having”, “including” and “comprising” used herein do not exclude the existence or addition of one or more elements or combinations.

[0033] The Lactobacillus salivarius used in each example was purchased from Shanghai Ruichu Biotechnology Co., Ltd. The strain number is SHMCC (SHBCC) D10168, and the number of viable bacteria is 2.7×10 9 CFU / g; Lactobacillus plantarum was purchased from Shanghai Lianmai Bioengineering Co., Ltd., with the product number ATCC337212, and the number of viable bacteria was 4.3×10 9 CFU / g.

[0034] The activation method of Lactobacillus plantarum used in each embodiment is as follows: 0.2 mL of sterile water is drawn with a sterile pipette, and 10 mg of purchased Lactobacillus plantarum powder is dissolved to obtain a bacterial solution; the raw material components of the culture medium are: 10 g of casein peptone, 10 g of beef extract, 10 g of yeast extract, 5 g of glucose, 1 g of Tween 80, 2 g of potassium hydrogen phosphate, 5 g of diamine citrate, 0.2 g of MgSO 4 7H 2 O, 0.05g 0.2g MnSO 4 7H 2 O, 1000mL distilled water, medium pH value is 6.5, sterilized at 121℃ for 15min, then the bacterial liquid is streaked into the medium, gently shaken to make the bacteria evenly distributed; the inoculated nutrient broth medium (Qingdao High-tech Industrial Park Haibo Biotechnology Co., Ltd.) is placed in an incubator and cultured at 37℃ for 48h; a single colony is selected from the nutrient broth medium and placed in the nutrient broth medium, and the nutrient broth medium is placed on a shaker at 180r / min for 24h to obtain an activated Lactobacillus plantarum bacterial liquid. The above activation method is also used for Lactobacillus salivarius.

[0035] Embodiment 1:

[0036] This embodiment provides a method for preparing a microecological composite preparation for improving the quality of boar semen, comprising the following steps:

[0037] Step 1, crushing cyperus oleifera meal and sieving through 120 meshes to obtain cyperus oleifera powder; adding 200g cyperus oleifera powder, 1g calcium carbonate, and 0.6g calcium hydrogen phosphate to 100mL sterile water, and then inoculating 0.5mL of activated bacterial solution of Lactobacillus germinalis and 1.5mL of activated bacterial solution of Lactobacillus salivarius respectively, stirring evenly, placing in a constant temperature incubator for culture at a temperature of 37°C and fermenting for 7 days to obtain cyperus oleifera fermentation product;

[0038] Step 2: Mix 200 g of cyperus oleifera fermentation product with 5 g of tryptone, 30 g of liver extract, 10 g of L-cysteine, and 20 g of fructose to obtain an inner layer mixture of the microecological composite preparation;

[0039] Step 3: preparing a barrier film colloid, using the barrier film colloid to coat the surface of the inner layer mixture of the microecological composite preparation with a barrier film; wherein the preparation method of the barrier film colloid includes:

[0040] S31, weighing 200g of modified konjac glucomannan, 100g of starch, 100g of soybean powder, and 20g of chitosan; wherein the preparation method of modified konjac glucomannan comprises:

[0041] S311, using 20g konjac glucomannan powder to prepare a konjac glucomannan solution with a concentration of 2wt%, sodium carbonate was added thereto to adjust the pH of the konjac glucomannan solution to 10, and after stirring for 2h, the solution was allowed to stand for 24h;

[0042] S312, add 5g guar gum to the konjac glucomannan solution, stir and mix to obtain a mixed solution;

[0043] S313, adding 2.5g of nano silicon dioxide powder and 0.3g of sodium tripolyphosphate to the mixed solution, stirring at a speed of 1200rpm and heating to 60°C, keeping the temperature for 2h, cooling to room temperature, evaporating and concentrating, and drying to obtain modified konjac glucomannan;

[0044] S32, first dispersing starch in 20000mL deionized water to obtain a starch dispersion with a concentration of 5mg / mL; heating the starch dispersion to 40°C, stirring continuously at a speed of 300rpm, adding modified konjac glucomannan, keeping the temperature for 20min and then cooling to room temperature, stopping stirring, and obtaining a gel system;

[0045] S33, adding soybean powder and chitosan to the gel system, dispersing them ultrasonically at a frequency of 60 kHz for 1 hour, and freeze-drying them at -20°C to obtain a barrier film colloid with a water content of 30%;

[0046] Step 4: Coat the outside of the barrier film with a surface film to obtain a microecological composite preparation; wherein, the preparation method of the surface film comprises: weighing 20g of xanthan gum, 20g of sodium carboxymethyl cellulose, 30g of cyperus oleifera meal, and 100g of starch by weight, adding the above substances into 10000mL of sterile water, stirring evenly to obtain a crude liquid, evaporating the crude liquid and coating it on the surface of the barrier film to obtain a surface film, and the preparation of the microecological composite preparation is completed.

[0047] Embodiment 2:

[0048] This embodiment provides a method for preparing a microecological composite preparation for improving the quality of boar semen. The difference between this embodiment and Example 1 is that the fermentation days in step 1 are 10 days, and the preparation methods and process parameters of steps 2 to 4 are the same as those in Example 1.

[0049] Embodiment 3:

[0050] This embodiment provides a method for preparing a microecological composite preparation for improving the quality of boar semen. The difference between this embodiment and Example 1 is that the fermentation days in step 1 are 14 days, and the preparation methods and process parameters of steps 2 to 4 are the same as those in Example 1.

[0051] Embodiment 4:

[0052] This embodiment provides a method for preparing a microecological composite preparation for improving the quality of boar semen, comprising the following steps:

[0053] Step 1, crushing cyperus oleifera meal and sieving through 120 meshes to obtain cyperus oleifera powder; adding 200g cyperus oleifera powder, 0.5g calcium carbonate, and 0.3g calcium hydrogen phosphate to 100mL sterile water, and then inoculating 0.5mL of activated bacterial solution of Lactobacillus germinalis and 2.0mL of activated bacterial solution of Lactobacillus salivarius respectively, stirring evenly, placing in a constant temperature incubator for cultivation at a temperature of 37°C and a fermentation period of 7 days to obtain cyperus oleifera fermentation product;

[0054] Step 2: Mix 300 g of Cyperus oleifera fermentation product with tryptone, 50 g of liver extract, 20 g of L-cysteine, and 50 g of fructose to obtain an inner layer mixture of the microecological composite preparation;

[0055] Step 3: preparing a barrier film colloid, using the barrier film colloid to coat the surface of the inner layer mixture of the microecological composite preparation with a barrier film; wherein the preparation method of the barrier film colloid includes:

[0056] S31, weighing 300g modified konjac glucomannan, 150g starch, 100g soybean powder, 40g chitosan; wherein, the preparation method of modified konjac glucomannan comprises:

[0057] S311, using 20g konjac glucomannan powder to prepare a konjac glucomannan solution with a concentration of 2wt%, sodium carbonate was added thereto to adjust the pH of the konjac glucomannan solution to 10, and after stirring for 2h, the solution was allowed to stand for 24h;

[0058] S312, adding 10g guar gum to the konjac glucomannan solution, stirring and mixing to obtain a mixed solution;

[0059] S313, adding 4g of nano-silicon dioxide powder and 0.4g of sodium tripolyphosphate to the mixed solution, stirring at a speed of 1200rpm and heating to 65°C, keeping the temperature for 2h, cooling to room temperature, evaporating and concentrating, and drying to obtain modified konjac glucomannan;

[0060] S32, first dispersing starch in 20000mL deionized water to obtain a starch dispersion with a concentration of 7.5mg / mL; heating the starch dispersion to 40°C, stirring continuously at a speed of 300rpm, adding modified konjac glucomannan, keeping the temperature for 20min and then cooling to room temperature, stopping stirring, and obtaining a gel system;

[0061] S33, adding soybean powder and chitosan to the gel system, dispersing them ultrasonically at a frequency of 60 kHz for 1 hour, and freeze-drying them at -20°C to obtain a barrier film colloid with a water content of 30%;

[0062] Step 4: Coat the outside of the barrier film with a surface film to obtain a microecological composite preparation; wherein, the preparation method of the surface film comprises: weighing 20g of xanthan gum, 20g of sodium carboxymethyl cellulose, 30g of cyperus oleifera meal, and 100g of starch by weight, adding the above substances into 10000mL of sterile water, stirring evenly to obtain a crude liquid, evaporating the crude liquid and coating it on the surface of the barrier film to obtain a surface film, and the preparation of the microecological composite preparation is completed.

[0063] Embodiment 5:

[0064] This embodiment provides a method for preparing a microecological composite preparation for improving the quality of boar semen, comprising the following steps:

[0065] Step 1, crushing cyperus oleifera meal and sieving through 120 meshes to obtain cyperus oleifera powder; adding 200g cyperus oleifera powder, 0.8g calcium carbonate, and 0.3g calcium hydrogen phosphate to 100mL sterile water, and then inoculating 0.5mL of activated bacterial solution of Lactobacillus germinalis and 2.0mL of activated bacterial solution of Lactobacillus salivarius respectively, stirring evenly, placing in a constant temperature incubator for culture at a culture temperature of 37°C and fermenting for 7 days to obtain cyperus oleifera fermentation product;

[0066] Step 2: Mix 500 g of Cyperus oleifera fermentation product with 5 g of tryptone, 30 g of liver extract, 20 g of L-cysteine, and 20 g of fructose to obtain an inner layer mixture of the microecological composite preparation;

[0067] Step 3: preparing a barrier film colloid, using the barrier film colloid to coat the surface of the inner layer mixture of the microecological composite preparation with a barrier film; wherein the preparation method of the barrier film colloid includes:

[0068] S31, weighing 400g modified konjac glucomannan, 200g starch, 100g soybean powder, 20g chitosan; wherein, the preparation method of modified konjac glucomannan comprises:

[0069] S311, using 20g konjac glucomannan powder to prepare a konjac glucomannan solution with a concentration of 2wt%, sodium carbonate was added thereto to adjust the pH of the konjac glucomannan solution to 10, and after stirring for 2h, the solution was allowed to stand for 24h;

[0070] S312, in the konjac glucomannan solution, add 12g guar gum, stir and mix, obtain a mixed solution;

[0071] S313, adding 5g of nano silicon dioxide powder and 0.45g of sodium tripolyphosphate to the mixed solution, stirring at a speed of 1200rpm and heating to 70°C, keeping the temperature for 2h, cooling to room temperature, evaporating and concentrating, and drying to obtain modified konjac glucomannan;

[0072] S32, first dispersing starch in 20000mL deionized water to obtain a starch dispersion with a concentration of 5mg / mL; heating the starch dispersion to 60°C, stirring continuously at a speed of 500rpm, adding modified konjac glucomannan, and cooling to room temperature after keeping warm for 20min, stopping stirring, and obtaining a gel system;

[0073] S33, adding soybean powder and chitosan to the gel system, dispersing them ultrasonically at a frequency of 60 kHz for 1 hour, and freeze-drying them at -20°C to obtain a barrier film colloid with a water content of 30%;

[0074] Step 4: Coat the outside of the barrier film with a surface film to obtain a microecological composite preparation; wherein, the preparation method of the surface film comprises: weighing 20g of xanthan gum, 20g of sodium carboxymethyl cellulose, 30g of cyperus oleifera meal, and 100g of starch by weight, adding the above substances into 10000mL of sterile water, stirring evenly to obtain a crude liquid, evaporating the crude liquid and coating it on the surface of the barrier film to obtain a surface film, and the preparation of the microecological composite preparation is completed.

[0075] Comparative Example 1:

[0076] This comparative example provides a method for preparing a microecological composite preparation for improving the quality of boar semen. The difference from Example 1 is that the fermentation day of step one is 0 days, and the preparation methods and process parameters of the remaining steps of this comparative example are the same as those of Example 1.

[0077] Comparative Example 2:

[0078] This comparative example provides a method for preparing a microecological composite preparation for improving the quality of boar semen. The difference from Example 4 is that the fermentation day of step one is 0 days, and the preparation methods and process parameters of the remaining steps of this comparative example are the same as those of Example 4.

[0079] Comparative Example 3:

[0080] This comparative example provides a method for preparing a microecological composite preparation for improving the quality of boar semen. The difference from Example 1 is that in step three, 200 g of modified konjac glucomannan is replaced with 200 g of unmodified konjac glucomannan, and the preparation methods and process parameters of the remaining steps of this comparative example are the same as those in Example 1.

[0081] Comparative Example 4:

[0082] This comparative example provides a method for preparing a microecological composite preparation for improving the quality of boar semen. The difference from Example 4 is that in step three, 300 g of modified konjac glucomannan is replaced with 300 g of unmodified konjac glucomannan, and the preparation methods and process parameters of the remaining steps of this comparative example are the same as those in Example 4.

[0083] The number of live bacteria in the microecological composite preparations prepared in Examples 1 to 5 and Comparative Examples 1 to 4 after gastric acid digestion was detected by a gastric acid simulation test. The reagent preparation method used in the gastric acid simulation test is as follows: 16.4 mL of 0.1 kg / L hydrogen chloride solution is dissolved in 800 mL of water, 10 g of pepsin (1000 NFu / mg) is added, the pH is adjusted to 2.0, and the volume is fixed to 1000 mL; Filter and sterilize with a 0.22 micron sterile filter for later use.

[0084] Conduct simulated gastric fluid test: add 0.5g of microecological compound preparation into sterile simulated gastric fluid, incubate anaerobically at 37℃ for 2h, dilute and spread the counts at 0 and 2h, and calculate the survival rate.

[0085] The viable bacteria counting method was used to detect the initial viable bacteria count of each species in the microecological composite preparations of each embodiment and comparative example, and the viable bacteria count of each species was detected after 100 days at 4°C. The results are shown in Table 1:

[0086] Table 1 The number of viable bacteria of each species in the microecological composite preparations of Examples 1 to 5 and Comparative Examples 1 to 4

[0087]

[0088] As can be seen from the above table, the microecological composite preparations prepared in Examples 1 to 5 all have excellent gastric acid resistance and can maintain high activity in a gastric acid simulated environment. Among them, since Comparative Examples 1 and Comparative Examples 2 are not fermented, they have excellent gastric acid resistance and also have more initial viable bacteria.

[0089] The raw materials were ground into powder according to the ratio of 65% corn, 13% wheat bran, 13% soybean meal, 5% cottonseed meal, 3% fish meal, 0.76% calcium hydrogen phosphate, and 0.24% salt to prepare boar feed; the boar feed was mixed with the proecological composite preparations prepared in Examples 1 to 5 and Comparative Examples 1 to 4 at a ratio of 5kg: 1t, and boar feed not formulated with the proecological composite preparation was used as a blank control.

[0090] 500 boars in good growth condition were selected and divided into 10 groups, 50 in each group. Each group of boars was fed with the microecological composite preparations prepared in Examples 1 to 5 and Comparative Examples 1 to 4, a mixture of boar feed, and a blank control boar feed. After 60 days of feeding, 10 boars were randomly selected from each group of boars, and semen was extracted. According to the detection method in GB 23238-2021 Normal temperature semen of breeding pigs, the ambient temperature was measured at 18 ° C, and the sperm motility, sperm density, number of forward-moving sperm, and sperm deformity rate of each group of boars were tested respectively, and Tables 2 to 11 were obtained:

[0091] Table 2 Sperm test data of boars fed with the microecological composite preparation prepared in Example 1

[0092]

[0093] Table 3 Sperm test data of boars fed with the microecological composite preparation prepared in Example 2

[0094]

[0095] Table 4 Sperm test data of boars fed with the microecological composite preparation prepared in Example 3

[0096]

[0097] Table 5 Sperm test data of boars fed with the microecological composite preparation prepared in Example 4

[0098]

[0099] Table 6 Sperm test data of boars fed with the microecological composite preparation prepared in Example 5

[0100]

[0101] Table 7 Sperm test data of boars fed with the microecological composite preparation prepared in Comparative Example 1

[0102]

[0103] Table 8 Sperm test data of boars fed with the microecological composite preparation prepared in Comparative Example 2

[0104]

[0105] Table 9 Sperm test data of boars fed with the microecological composite preparation prepared in Comparative Example 3

[0106]

[0107] Table 10 Sperm test data of boars fed with the microecological composite preparation prepared in Comparative Example 4

[0108]

[0109] Table 11 Sperm test data of boars fed blank control feed

[0110]

[0111] It can be seen from the above tables that after feeding the boar feed prepared with the microecological composite preparations of Examples 1 to 4, the boars' sperm motility, sperm density, number of forward-moving sperm, and sperm deformity rate were significantly better than those of Comparative Examples 1 to 4 and the blank control group, and the semen quality was significantly improved.

[0112] The number of devices and processing scales described here are used to simplify the description of the present invention. Applications, modifications and variations of the present invention will be obvious to those skilled in the art.

[0113] Although the embodiments of the present invention have been disclosed as above, they are not limited to the applications listed in the specification and the implementation modes, and they can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to the specific details and the examples shown and described herein.

Claims

1. A method for preparing a microecological composite preparation for improving the quality of boar semen, characterized in that: The following steps are involved: Step 1: fermenting cyperus oleifera meal with a mixed bacteria of Lactobacillus plantarum and Lactobacillus salivarius at an inoculation volume ratio of 1:2-4 to obtain cyperus oleifera fermented product; Step 2: mixing the fermented product of Cyperus oleifera with tryptone, liver extract, L-cysteine ​​and fructose to obtain an inner layer mixture of the microecological composite preparation; Step 3: Preparation of barrier film colloid: S31, weighing 2-4 parts of modified konjac glucomannan, 1-2 parts of starch, 1-2 parts of soybean flour, and 0.2-0.5 parts of chitosan by weight; S32, first disperse the starch in 200-300 parts of deionized water to obtain a starch dispersion with a concentration of 2-10 mg / mL; heat the starch dispersion to 40-70° C., and continue stirring at a speed of 200-500 rpm, Add modified konjac glucomannan, keep warm for 10-20 minutes, then cool to room temperature, stop stirring, and obtain a gel system; S33, adding soybean powder and chitosan to the gel system, ultrasonically dispersing at a frequency of 60-80 kHz for 1-3 hours, and freeze-drying at -30--10°C to obtain a barrier film colloid with a water content of 25%-33%; The preparation method of modified konjac glucomannan comprises: S311, using konjac glucomannan powder to prepare a konjac glucomannan solution with a concentration of 1.5-2wt%, adding sodium carbonate to adjust the pH of the konjac glucomannan solution to 9-10, stirring for 2h, and leaving it to stand for 24-36h; S312, in konjac glucomannan solution, add guar gum, make the concentration of guar gum be 0.5 ~ 1wt%, stir and mix, obtain mixed solution; S313, adding nano silicon dioxide powder and sodium tripolyphosphate with a particle size of 20-80 nm and a D90 particle size of less than 55 nm to the mixed solution, stirring at a speed of 600-1200 rpm and heating to 50-80° C., keeping warm for 2-5 hours, cooling to room temperature, evaporating and concentrating, and drying to obtain modified konjac glucomannan; the mass ratio of konjac glucomannan powder, nano silicon dioxide powder, and sodium tripolyphosphate is 4:0.5-1:0.05-0.1; using a barrier film colloid to coat the surface of the inner layer mixture of the microecological composite preparation with a barrier film; Step 4: Coat the surface film on the outside of the barrier film to obtain a microecological composite preparation.

2. The method for preparing the microecological composite preparation for improving the quality of boar semen according to claim 1, characterized in that: The specific method of step one includes: S11, selecting bacterial strains for activation respectively to obtain activated bacterial strains; S12, crushing cyperus chinensis meal and sieving through 120 mesh to obtain cyperus chinensis powder; adding calcium carbonate and calcium hydrogen phosphate to the cyperus chinensis powder, then inoculating activated bacteria, stirring evenly, placing in a constant temperature incubator for culture at a temperature of 37° C., and fermenting for 7 to 16 days; wherein the mass ratio of calcium carbonate, calcium hydrogen phosphate and cyperus chinensis powder is 1 to 5:1 to 3:1000 to 2000.

3. The method for preparing the microecological composite preparation for improving the quality of boar semen according to claim 1, characterized in that: In the step 2, the mass ratio of the cyperus oleifera fermentation product to tryptone, liver extract, L-cysteine, and fructose is 2-5: 0.05-0.1: 0.3-1: 0.1-0.2: 0.2-1.

4. The method for preparing the microecological composite preparation for improving boar semen quality according to claim 1, characterized in that: The preparation method of the surface film comprises: weighing 0.2-0.6 parts of xanthan gum, 0.2-0.4 parts of sodium carboxymethyl cellulose, 0.3-0.8 parts of cyperus oleifera meal, and 1-1.5 parts of starch by weight, adding them into 10-12 parts of sterile water, stirring evenly to obtain a crude liquid, evaporating the crude liquid and coating it on the surface of the barrier film to obtain the surface film.

5. A microecological composite preparation for improving the quality of boar semen, characterized in that: The microecological composite preparation for improving the quality of boar semen is prepared by the preparation method of any one of claims 1 to 4.

6. An application of the microecological composite preparation for improving the quality of boar semen as claimed in claim 5, characterized in that: The microecological composite preparation is used as an additive in boar feed, and the mixing ratio of the microecological composite preparation to feed is 0.003-0.005:1; wherein, the storage condition of the microecological composite preparation is to store it in a dry place away from light at 2-4°C, and the storage time shall not exceed 7 days.

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