CN109874920B

CN109874920B - Compound microbial feed additive and preparation method thereof

Info

Publication number: CN109874920B
Application number: CN201910278906.2A
Authority: CN
Inventors: 王健; 刘坤; 唐跃辉; 齐静; 徐克东
Original assignee: Zhoukou Normal University
Current assignee: Liaoning Qixing Biotechnology Co ltd
Priority date: 2019-04-09
Filing date: 2019-04-09
Publication date: 2022-01-11
Anticipated expiration: 2039-04-09
Also published as: CN109874920A

Abstract

本发明公开了一种复合微生物饲料添加剂，由以下重量份的原料制备而成：乳酸菌发酵液30‑50份、胶冻样芽孢杆菌发酵液10‑15份、长枝木霉发酵液10‑15份、小球藻藻液20‑30份。本发明还提供了一种复合微生物饲料添加剂的制备方法。本发明提供的复合微生物饲料添加剂能改善仔猪肠道功能，提高断奶仔猪日增重量和饲料转化效率，还具有显著的抗菌、抗病毒的作用，可以显著的提高仔猪的免疫力，有效的预防仔猪断奶后出现腹泻的现象，具备广泛应用前景。The invention discloses a composite microbial feed additive, which is prepared from the following parts by weight of raw materials: 30-50 parts of lactic acid bacteria fermentation liquid, 10-15 parts of Bacillus jelly-like fermentation liquid, and 10-15 parts of Trichoderma longbranchus fermentation liquid parts, 20-30 parts of Chlorella algal liquid. The invention also provides a preparation method of the compound microorganism feed additive. The compound microbial feed additive provided by the invention can improve the intestinal function of piglets, increase the daily weight gain and feed conversion efficiency of weaned piglets, and also have obvious antibacterial and antiviral effects, can significantly improve the immunity of piglets, and effectively prevent piglets. The phenomenon of diarrhea after weaning has a wide application prospect.

Description

Compound microbial feed additive and preparation method thereof

Technical Field

The invention belongs to the technical field of animal feed processing, and particularly relates to a compound microbial feed additive and a preparation method thereof.

Background

The microbial feed additive is a special feed additive, promotes the growth of cultured animals through the physiological metabolism of microbial strains, reduces the use of medicaments, improves the quality of the cultured animals, and realizes green culture. Due to the excellent properties of the microbial feed additives, more and more researchers have started the research, and especially the application of the microbial feed additives in the pig industry has become a research hotspot at present.

In the pig raising industry, in order to reduce epidemic diseases spread from sows to piglets, piglets of 3-5 weeks are weaned, but the weaned piglets often suffer from diarrhea due to incomplete self-digestion function and low immunity, so the normal growth and development of the piglets are influenced, and the piglets can die if the self-digestion function is severe, and the popularization and the application of early weaning technology of the piglets are restricted by the occurrence of the condition.

The main measures adopted at present for the condition are to add antibiotics into feed of weaned pigs to improve the immunity of the piglets, prevent diarrhea of the piglets and promote growth, but the antibiotics are easy to generate drug-resistant bacteria after long-term use, and the bacteria easily influence the health of human beings through food chains. Therefore, it is necessary to apply the microbial feed additive to the breeding process of the weaned piglets to solve the problem that the weaned piglets are easy to diarrhea at present.

Disclosure of Invention

The invention provides a compound microbial feed additive, which solves the problems that in the prior art, antibiotics are used for improving the immunity of piglets, drug-resistant bacteria are easy to generate in piglet diarrhea prevention, and the drug-resistant bacteria are easy to influence the health of human beings through a food chain.

The invention aims to provide a compound microbial feed additive which is prepared from the following raw materials in parts by weight: 30-50 parts of lactobacillus fermentation liquor, 10-15 parts of bacillus mucilaginosus fermentation liquor, 10-15 parts of trichoderma longibrachiatum fermentation liquor and 20-30 parts of chlorella liquid.

Wherein the effective viable count of the lactobacillus fermentation liquor and the bacillus mucilaginosus fermentation liquor is 1.0 multiplied by 10¹⁰-3.0×10¹⁰CFU/mL; the spore amount in the Trichoderma longibrachiatum fermentation liquor is 1.0 multiplied by 10⁸-1.5×10⁹Per mL; the chlorella solution has a concentration of 1.0 × 10⁵-2.0×10⁵one/mL.

The second purpose of the invention is to provide a preparation method of the compound microorganism feed additive, which comprises the following steps:

step 1, respectively preparing lactobacillus seed liquid and bacillus mucilaginosus seed liquid;

step 2, preparing lactobacillus fermentation liquor

Inoculating the lactobacillus seed liquid into a YPD liquid culture medium, and then culturing at 35 +/-2 ℃ at an oscillation speed of 200r/min to the end of logarithm to obtain lactobacillus fermentation liquid after the culture is finished;

step 3, preparing the bacillus mucilaginosus fermentation liquor

Drying persimmon peel, and grinding into powder to obtain persimmon peel powder;

drying persimmon stone, and grinding into powder to obtain persimmon stone powder;

mixing persimmon peel powder, persimmon stone powder and beef extract peptone liquid culture medium according to the ratio of 1-2: 2-5: 20 to obtain the bacillus mucilaginosus culture medium;

inoculating a bacillus mucilaginosus seed solution into the bacillus mucilaginosus culture medium, uniformly mixing, and then culturing at 28 +/-2 ℃ at an oscillation speed of 200r/min to the late logarithmic phase to obtain a bacillus mucilaginosus fermentation solution;

step 4, preparing trichoderma longibrachiatum fermentation liquor

Inoculating the trichoderma longibrachiatum into a PDB liquid culture medium according to the inoculation amount of 5% by mass, and culturing for 5 days at the temperature of 30 +/-2 ℃ to obtain a trichoderma longibrachiatum fermentation product; then eluting trichoderma longibrachiatum spores on the trichoderma longibrachiatum fermentation product by using distilled water to obtain trichoderma longibrachiatum fermentation liquor;

step 5, preparing chlorella liquid

Inoculating chlorella into BG11 culture medium according to an inoculation amount of 5% by mass, and culturing at 25 + -1 deg.C under illumination intensity of 5000lux and light-dark ratio of 12 h: culturing for 12h to logarithmic growth phase to obtain chlorella solution;

step 6, weighing 30-50 parts of lactobacillus fermentation liquor, 10-15 parts of bacillus mucilaginosus fermentation liquor, 10-15 parts of trichoderma longibrachiatum fermentation liquor and 20-30 parts of chlorella liquid according to parts by weight, uniformly mixing to obtain mixed fermentation liquor, and carrying out ventilation drying on the mixed fermentation liquor at 35-40 ℃ until the water content is less than or equal to 10% to obtain the compound microorganism feed additive.

Preferably, the inoculation amount of the lactobacillus seed solution and the bacillus mucilaginosus seed solution is 10ml of seed solution per kilogram of matrix.

Preferably, the granularity of the persimmon peel powder and the granularity of the persimmon kernel powder are both 10-20 meshes.

Compared with the prior art, the invention has the beneficial effects that:

1) the invention adopts a plurality of microorganisms for matching use, and the synergistic effect is utilized to play a role; the lactobacillus can promote the absorption of the piglets on nutrient substances in the feed, and also has the effects of increasing intestinal beneficial flora and improving the gastrointestinal tract function of the piglets; the bacillus mucilaginosus can generate various hormone substances, biological enzymes, amino polysaccharides, proteins and amino acid substances in intestinal tracts of piglets, and can promote the growth and development of the piglets and enhance the antiviral capacity of the piglets; the trichoderma longibrachiatum can grow and reproduce by taking pathogenic bacteria mycelia as nutrition, can induce the trichoderma longibrachiatum to generate a plurality of extracellular cell wall hydrolases, disintegrates pathogenic bacteria cells by the synergistic enzyme dissolving effect, can generate strong antibacterial active substances, and can inhibit the growth of pathogenic bacteria in intestinal tracts of piglets; the chlorella is rich in nutrition, contains natural proteins, amino acids, unsaturated fatty acids, vitamins, mineral substances and the like, can provide comprehensive nutrition for the growth of piglets, has a certain antibacterial effect, and prevents the growth and reproduction of pathogenic bacteria in intestinal tracts of piglets under the combined action of multiple mechanisms of bacillus mucilaginosus, trichoderma longibrachiatum and chlorella, thereby enhancing the disease-resistant effect of the piglets.

2) The compound microbial feed additive provided by the invention has remarkable antibacterial and antiviral effects, can remarkably improve the immunity of piglets and effectively prevent the phenomenon of diarrhea after the piglets are weaned; can also improve the intestinal function of the piglets, improve the daily weight gain and the feed conversion efficiency of the weaned piglets and have wide application prospect.

Detailed Description

In order to make the technical solutions of the present invention better understood and enable those skilled in the art to practice the present invention, the following embodiments are further described, but the present invention is not limited to the following embodiments.

The lactic acid bacteria, the bacillus mucilaginosus, the trichoderma longibrachiatum and the chlorella used in the following examples are all the existing strains which can be purchased in the microorganism strain preservation management center, do not relate to the development of new strains, and only relate to the application of the existing strains. In addition, the lactobacillus used in the following examples is specifically a lactobacillus which has a preservation number of CCTCC NO: a strain of M2017382; the bacillus mucilaginosus is a strain with the preservation number of CGMCC No.16005 of China general microbiological culture Collection center; the trichoderma longibrachiatum is specifically a strain with the preservation number of CGMCC No.8723 of China Committee for culture Collection of microorganisms; the chlorella is preserved in China center for type culture collection with the preservation number of CCTCC NO: m209256;

in the examples, both the lactic acid bacteria seed solution and the Bacillus mucilaginosus seed solution were obtained by culturing according to a conventional method, and the test methods described in the following examples are according to a conventional method unless otherwise specified.

Example 1

A composite microbial feed additive is prepared from the following raw materials in parts by weight: 30 parts of lactobacillus fermentation liquor, 15 parts of bacillus mucilaginosus fermentation liquor, 10 parts of trichoderma longibrachiatum fermentation liquor and 20 parts of chlorella liquid;

wherein the effective viable count of lactobacillus fermentation broth and Bacillus mucilaginosus fermentation broth is 2.6 × 10¹⁰CFU/mL; the spore amount in the fermentation liquid of the trichoderma longibrachiatum is 1.0 multiplied by 10⁸Per mL; the chlorella solution has a concentration of 1.8 × 10⁵one/mL.

The preparation method comprises the following steps:

step 2, preparing lactobacillus fermentation liquor

step 3, preparing the bacillus mucilaginosus fermentation liquor

Drying persimmon peel, grinding into powder, and sieving with a 10-mesh sieve to obtain persimmon peel powder;

drying persimmon kernels, grinding the dried persimmon kernels into powder, and sieving the powder with a 10-mesh sieve to obtain persimmon kernel powder;

mixing persimmon peel powder, persimmon stone powder and beef extract peptone liquid culture medium according to the proportion of 1: 2: 20 to obtain the bacillus mucilaginosus culture medium;

step 4, preparing trichoderma longibrachiatum fermentation liquor

step 5, preparing chlorella liquid

Inoculating chlorella into BG11 culture medium according to an inoculation amount of 5% by mass, and culturing at 25 + -1 deg.C under illumination intensity of 5000lux and light-dark ratio of 12 h: culturing for 12h to logarithmic phase to obtain concentration of 1.8 × 10⁵Per mL of chlorella algae solution;

step 6, weighing 30-50 parts of lactobacillus fermentation liquor, 10-15 parts of bacillus mucilaginosus fermentation liquor, 10-15 parts of trichoderma longibrachiatum fermentation liquor and 20-30 parts of chlorella liquid according to parts by weight, uniformly mixing to obtain mixed fermentation liquor, and carrying out ventilation drying on the mixed fermentation liquor at 35-40 ℃ until the water content is 5.3%, thus obtaining the compound microorganism feed additive.

Example 2

A composite microbial feed additive is prepared from the following raw materials in parts by weight: 40 parts of lactobacillus fermentation liquor, 10 parts of bacillus mucilaginosus fermentation liquor, 12 parts of trichoderma longibrachiatum fermentation liquor and 25 parts of chlorella liquid;

wherein the effective viable count of lactobacillus fermentation broth and Bacillus mucilaginosus fermentation broth are all1.0×10¹⁰CFU/mL; the spore amount in the fermentation liquid of the trichoderma longibrachiatum is 1.0 multiplied by 10⁹Per mL; the chlorella solution has a concentration of 1.5 × 10⁵one/mL.

The preparation method is the same as that of example 1, except that the formula of example 1 is changed to that of example 2, and the preparation method of the bacillus mucilaginosus fermentation liquid is as follows:

drying persimmon peel, grinding into powder, and sieving with a 20-mesh sieve to obtain persimmon peel powder;

drying persimmon kernels, grinding the dried persimmon kernels into powder, and sieving the powder with a 20-mesh sieve to obtain persimmon kernel powder;

mixing persimmon peel powder, persimmon stone powder and beef extract peptone liquid culture medium according to the proportion of 1: 5: 20 to obtain the bacillus mucilaginosus culture medium;

inoculating the bacillus mucilaginosus seed solution into the bacillus mucilaginosus culture medium, uniformly mixing, and then culturing at 28 +/-2 ℃ at the oscillation speed of 200r/min to the late logarithmic phase to obtain the bacillus mucilaginosus fermentation solution. The moisture content of the composite microbial feed additive prepared in example 2 was 10%.

Example 3

A composite microbial feed additive is prepared from the following raw materials in parts by weight: 50 parts of lactobacillus fermentation liquor, 12 parts of bacillus mucilaginosus fermentation liquor, 15 parts of trichoderma longibrachiatum fermentation liquor and 30 parts of chlorella liquid;

wherein the effective viable count of lactobacillus fermentation broth and Bacillus mucilaginosus fermentation broth is 3.0 × 10¹⁰CFU/mL; the spore amount in the fermentation liquid of the trichoderma longibrachiatum is 1.5 multiplied by 10⁹Per mL; the chlorella solution has a concentration of 1.0 × 10⁵one/mL.

The preparation method is the same as that of example 1, except that the formula of example 1 is changed to that of example 3, and the preparation method of the bacillus mucilaginosus fermentation liquid is as follows:

mixing persimmon peel powder, persimmon stone powder and beef extract peptone liquid culture medium according to the ratio of 2: 2: 20 to obtain the bacillus mucilaginosus culture medium;

inoculating the bacillus mucilaginosus seed solution into the bacillus mucilaginosus culture medium, uniformly mixing, and then culturing at 28 +/-2 ℃ at the oscillation speed of 200r/min to the late logarithmic phase to obtain the bacillus mucilaginosus fermentation solution. The moisture content of the composite microbial feed additive prepared in example 3 was 3.8%.

Example 4

wherein the effective viable count of lactobacillus fermentation broth and Bacillus mucilaginosus fermentation broth is 1.8 × 10¹⁰CFU/mL; the spore amount in the fermentation liquid of the trichoderma longibrachiatum is 3.5 multiplied by 10⁸Per mL; the chlorella solution has a concentration of 2.0 × 10⁵one/mL.

The preparation method is the same as that of example 1, except that the preparation method of the bacillus mucilaginosus fermentation liquid is as follows:

mixing persimmon peel powder, persimmon stone powder and beef extract peptone liquid culture medium according to the ratio of 2: 5: 20 to obtain the bacillus mucilaginosus culture medium;

inoculating the bacillus mucilaginosus seed solution into the bacillus mucilaginosus culture medium, uniformly mixing, and then culturing at 28 +/-2 ℃ at the oscillation speed of 200r/min to the late logarithmic phase to obtain the bacillus mucilaginosus fermentation solution. The moisture content of the composite microbial feed additive prepared in example 4 was 8.7%.

In order to verify the effects of the persimmon peel powder and the persimmon stone powder, the following comparative examples were provided.

Comparative example 1

The preparation method of the bacillus mucilaginosus fermentation liquor comprises the following steps:

mixing persimmon peel powder and beef extract peptone liquid culture medium according to the ratio of 1: 20 to obtain the bacillus mucilaginosus culture medium;

inoculating the bacillus mucilaginosus seed solution into the bacillus mucilaginosus culture medium, uniformly mixing, and then culturing at 28 +/-2 ℃ at the oscillation speed of 200r/min to the late logarithmic phase to obtain the bacillus mucilaginosus fermentation solution.

Comparative example 2

mixing persimmon stone powder and a beef extract peptone liquid culture medium according to the ratio of 2: 20 to obtain the bacillus mucilaginosus culture medium;

Comparative example 3

inoculating the bacillus mucilaginosus seed solution into a beef extract peptone liquid culture medium, uniformly mixing, and then culturing at 28 +/-2 ℃ at the oscillation speed of 200r/min to the late logarithmic phase to obtain the bacillus mucilaginosus fermentation solution.

The effective viable count of Bacillus mucilaginosus in the Bacillus mucilaginosus fermented liquid obtained in example 1 and comparative examples 1-3 was determined, and is shown in Table 1.

TABLE 1 effective viable count in Bacillus mucilaginosus fermentation broth

Method	Example 1	Comparative example 1	Comparative example 2	Comparative example 3
					Effective viable count (10)¹⁰CFU/ml)	2.6	1.2	1.8	0.5

As can be seen from table 1, the effective viable count of bacillus mucilaginosus in the bacillus mucilaginosus fermentation liquid of comparative example 3 is the lowest, and the other groups are higher, which indicates that the addition of persimmon peel powder and persimmon stone powder in the fermentation medium is helpful for increasing the growth of bacillus mucilaginosus, while the common beef extract peptone liquid medium does not contain persimmon peel powder and persimmon stone powder, so the effective viable count is lower;

the persimmon peel powder and the persimmon core powder are added in the embodiment 1 and the comparative examples 1-2, so that the number of effective viable bacteria is higher, the effect is better than that of a beef extract peptone liquid culture medium, and the persimmon peel powder and the persimmon core powder play an important role in promoting the growth of bacillus mucilaginosus; by comparing the data of example 1, comparative example 1 and comparative example 2, we find that the effect of the persimmon kernel powder is better than that of the persimmon peel powder, and the effect of the matching use of the persimmon peel powder and the persimmon kernel powder is better, which indicates that the persimmon peel powder and the persimmon kernel powder can be synergistic.

The test results show that the culture medium provided by the embodiment of the invention has the effect of obviously increasing the effective viable count of the bacillus mucilaginosus.

In order to further illustrate the effect of the invention, the invention is also provided with the following comparative examples:

comparative example 4

The microbial feed additive only contains lactobacillus fermentation liquor.

Comparative example 5

A compound microorganism feed additive has the same formula as that of example 1, except that Bacillus mucilaginosus fermentation liquor is not added in the formula of comparative example 1, and the preparation method does not contain the preparation step of the Bacillus mucilaginosus fermentation liquor.

Comparative example 6

A compound microorganism feed additive has the same formula as that of the feed additive in the embodiment 1, and is different in that the formula of the comparative example 1 is not added with Trichoderma longibrachiatum fermentation liquor, and the preparation method does not contain the preparation step of the Trichoderma longibrachiatum fermentation liquor.

Comparative example 7

A compound microorganism feed additive has the same formula as that of example 1, except that the formula of comparative example 1 does not contain chlorella solution, and the preparation method does not contain the preparation step of chlorella solution.

In the examples and comparative examples of the present invention, the inoculation amounts of the lactic acid bacteria seed solution and the bacillus mucilaginosus seed solution were 10ml of seed solution per kg of substrate.

Hereinafter, only the performance test of the microbial feed additives prepared in example 1 and comparative examples 4 to 7 will be described to illustrate the effects of the present invention.

1. Test object

The experimental animals are selected from 120 healthy piglets which are 28 +/-1 day old and 5.5-6.5kg in average weight and are bred under the same breeding condition, the healthy piglets are randomly divided into 6 groups, 20 piglets in each group have no obvious difference among the groups. The test group was 1 group, and the microbial feed additive prepared in example 1 plus the general feed was fed; the control group is 4 groups, and the microbial feed additive and the common feed prepared in the comparative examples 1 to 4 are fed respectively; the drug control group was fed with normal feed plus antibiotics (10 mg/kg of colistin sulfate plus 25mg/kg of lidomycin). All piglets were infused with 5mL sheep red blood cell suspension with mass concentration of 1.0% via ear vein on the day of 28 days of age, and the test period was 14 days.

The mass ratio of the addition amount of the microbial feed additive to the addition amount of the common feed in each group is 1: 99, the mass ratio of the antibiotics to the common feed is also 1: 99 and the normal feed is a 10% piglet compound premix feed purchased from Anhui Xinhui animal science and technology development Limited.

2. Test method

The piglets of each group are fed by high bed leakage, and the pigsty is disinfected according to the conventional method so as to be fed and drunk freely. Before, after 7d and 14d, a small amount of blood was collected from 9 groups of living bodies via the auricular vein and 8-10mL (anticoagulated by adding 1% heparin sodium) via the jugular vein respectively under fasting condition, and then the percentage of neutrophils and peripheral blood lymphocytes, the antibody titer of sheep erythrocytes, the content of serum gamma-globulin and the proliferation rate of monocytes in peripheral blood were measured.

Wherein, the percentage of the neutrophils and the peripheral blood lymphocytes is measured by adopting the auricular venous blood to carry out conventional white blood cell differential counting;

the sheep red blood cell antibody titer is determined by adopting auricular venous blood and using a conventional micro-method hemagglutination test to determine the piglet sheep red blood cell antibody titer (the sheep red blood cell concentration is 1.0%);

the method for measuring the content of the gamma-globulin in the serum comprises the following steps: separating blood serum from auricular venous blood, separating each protein component in the blood serum by an acetate fiber thin film electrophoresis method, measuring an OD value on a spectrophotometer at 620nm after dyeing and elution, and measuring the relative content of gamma-globulin;

the method for measuring the proliferation rate of monocytes in peripheral blood is as follows: diluting Hank's solution of heparin sodium anticoagulated blood, adding Ficoll separating medium (GE Healthcare), and horizontally centrifuging at 2000r/min for 30 min; sucking the mononuclear cell interface layer, washing with Hank's solution for 2 times, and adjusting the cell concentration to 2 × 10 with RPMI 1640 complete culture solution⁶Perml, 40 well sterile cell culture plates were added, 0.1mL of cell suspension was added per well, and R with or without PHA was addedPMI1640 complete medium (final concentration 10. mu.g/mL) was used as a control for stimulated or spontaneous transformation, 3 wells per sample. Placing at 37 ℃ and 5% CO₂The culture was carried out in an incubator for 66 hours, and then MTT solution (prepared in PBS, pH 7.2) was added to give a final concentration of 0.5 mg/mL. The culture was continued for 6h, followed by addition of 20% SDS solution (containing 50% dimethylformamide, pH 4.7) and incubation at 37 ℃ for 12h, and OD was measured at 570nm on a spectrophotometer.

During the trial period, the diarrhea rate of each group of piglets was recorded every day during the trial period, and the piglets were weighed on an empty stomach on the 14 th morning of the trial period, and the pig weight gain and the feed-weight ratio were calculated.

3. Data processing

Statistical processing is carried out by using SPSS12.0 statistical software, data of measured data are expressed, single-factor variance analysis is carried out by comparing average numbers of a plurality of samples, and counting data are x²And (6) checking.

4. Test results

The test results are shown in table 2.

TABLE 2 Effect on percentage of neutrophils and peripheral blood lymphocytes in piglets/%

As can be seen from Table 2, the microbial feed additive provided in example 1 of the present invention can significantly increase neutrophils in the blood system of piglets and significantly reduce lymphocytes, and the effect is equivalent to that of antibiotics.

TABLE 3 Effect on piglet serum sheep red blood cell antibody titers (log2)

As can be seen from Table 3, the feed additive provided in example 1 of the present invention can significantly improve the immune response of piglets, and the effect is equivalent to that of antibiotics.

TABLE 4 Effect on piglet serum Gamma-globulin content (OD value)

As can be seen from Table 4, the feed additive provided by the embodiment 1 of the invention can obviously enhance the immunity of piglets, and the effect is equivalent to that of antibiotics.

TABLE 5 Effect on the proliferation Rate (OD value) of monocytes in peripheral blood of piglets

As can be seen from table 5, the peripheral blood mononuclear cell proliferation rate was significantly reduced by using the antibiotics, but the examples and comparative examples according to the present invention had no significant effect on the peripheral blood mononuclear cell proliferation rate.

TABLE 6 Effect on piglet Productivity

As can be seen from table 6, after the compound microbial feed additive of example 1 is fed, the daily feed intake, daily gain and diarrhea control rate of piglets within 2 weeks are all equivalent to those of the drug control group, which indicates that the compound microbial feed additive provided by the invention can significantly improve the intestinal development and health of piglets, relieve the stress reaction of piglets after weaning, effectively improve the feed intake and body mass of piglets, and effectively solve the problem of diarrhea of piglets.

While the present invention has been described with respect to preferred embodiments, additional variations and modifications will occur to those embodiments once the basic inventive concepts are known to those skilled in the art. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.

It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.

Claims

1. a compound microorganism feed additive, is characterized in that, is prepared from the raw material of following parts by weight: 30-50 parts of lactic acid bacteria fermentation liquid, 10-15 parts of jelly-like Bacillus fermentation liquid, 10 parts of Trichoderma longbranchus fermentation liquid -15 parts, 20-30 parts of Chlorella algae liquid.

Wherein, the number of effective viable bacteria in the lactic acid bacteria fermentation liquid and the Bacillus jelly-like fermentation liquid are both 1.0×10 ¹⁰ -3.0×10 ¹⁰ CFU/mL; the amount of spores in the Trichoderma longbranchus fermentation liquid is 1.0 ×10 ⁸ -1.5×10 ⁹ cells/mL; the concentration of the Chlorella algal liquid is 1.0×10 ⁵ -2.0×10 ⁵ cells/mL;

The compound microbial feed additive is prepared according to the following steps:

Step 1, prepare lactic acid bacteria seed liquid and jelly-like bacillus seed liquid respectively;

Step 2, prepare lactic acid bacteria fermentation broth

The lactic acid bacteria seed liquid is inoculated into the YPD liquid medium, and then cultivated to the end logarithmic stage at an oscillation speed of 200 r/min at 35 ± 2 °C, and the lactic acid bacteria fermentation broth is obtained after the cultivation is completed;

Step 3, Preparation of Bacillus jelly-like fermentation broth

The persimmon skin is dried and ground into powder to obtain persimmon skin powder;

The persimmon core is dried and ground into powder to obtain persimmon core powder;

The persimmon skin powder, the persimmon core powder and the beef extract peptone liquid medium are uniformly mixed according to the mass ratio of 1-2:2-5:20 to obtain a jelly-like Bacillus culture medium;

Inoculate the jelly-like Bacillus seed solution into the jelly-like Bacillus culture medium, mix well, and then culture at 28±2°C with a shaking speed of 200 r/min to the end of the logarithmic phase to obtain a jelly-like Bacillus germinative solution ;

Step 4, prepare Trichoderma longbranchs fermentation broth

Inoculate Trichoderma longbranchs in the PDB liquid medium according to the mass fraction of 5% of the inoculum, and cultivate at 30±2°C for 5 days to obtain Trichoderma longbranchs fermentation product; The Trichoderma longbranchs spores obtain Trichoderma longbranchs fermentation broth;

Step 5, prepare Chlorella algal liquid

Chlorella was inoculated into BG11 medium according to the mass fraction of 5% inoculum, and cultivated to the logarithmic growth phase under the conditions of 25±1°C, light intensity of 5000 lux, and light-dark ratio of 12h: 12h to obtain Chlorella. algal fluid;

Step 6, weigh 30-50 parts of lactic acid bacteria fermentation liquid, 10-15 parts of jelly-like Bacillus fermentation liquid, 10-15 parts of Trichoderma longbranchus fermentation liquid, 20-30 parts of Chlorella algae liquid by weight, mix uniformly, to obtain a mixed fermentation liquid, and the mixed fermentation liquid is ventilated and dried at 35-40° C. to a moisture content of ≤10% to obtain the compound microbial feed additive.

2 . The compound microbial feed additive according to claim 1 , wherein the inoculum amount of the lactic acid bacteria seed liquid and the jelly-like Bacillus seed liquid is 10 ml of seed liquid per kilogram of substrate. 3 .

3 . The compound microbial feed additive according to claim 1 , wherein the particle sizes of the persimmon skin powder and the persimmon core powder are both 10-20 meshes. 4 .