阅读说明：本技术 一种微生态微胶囊包被制剂及其制备方法 (Micro-ecological microcapsule coating preparation and preparation method thereof ) 是由 汪安国 田玉虎 李凤华 于 2021-09-26 设计创作，主要内容包括：本发明公开一种微生态微胶囊包被制剂及其制备方法,属于饲料添加剂技术领域。所述微生态微胶囊包被制剂包含动物双歧杆菌、干酪乳杆菌和布拉迪酵母的活菌,以及这三种菌的代谢产物。本发明所述微生态微胶囊包被型制剂能够有效避免菌通过饲料制粒以及胃液、胰液、胆汁等消化后和常温长期放置的损失,发挥其有效作用,延长了保质期。(The invention discloses a micro-ecological microcapsule coating preparation and a preparation method thereof, belonging to the technical field of feed additives. The micro-ecological microcapsule coating preparation comprises live bacteria of animal bifidobacterium, lactobacillus casei and saccharomyces boulardii and metabolites of the three bacteria. The micro-ecological microcapsule coated preparation can effectively avoid the loss of bacteria after feed granulation and digestion of gastric juice, pancreatic juice, bile and the like and after long-term storage at normal temperature, play the effective role of the bacteria and prolong the shelf life.)

1. A preparation method of a microecological bactericide is characterized by comprising the following steps:

(1) aerobic fermentation

According to 2% > EInoculating the Saccharomyces boulardii seed liquid to a mixed culture medium with 3 percent of inoculation amount, and carrying out aerobic fermentation at the fermentation temperature of 25-28 ℃, the pressure of 0.04-0.06 MPa, the rotation speed of 180-200 r/min and the ventilation volume of 200-250 m³Fermenting for 18-24 hours;

(2) anaerobic fermentation

Adding 1-3 wt% of sugar into the fermentation system in the step (1), and adjusting the pH value to 7.0; inoculating the bifidobacterium animalis seed liquid and the lactobacillus casei seed liquid to a fermentation system according to the inoculation amounts of 1-2% and 2-3% respectively, and carrying out anaerobic fermentation at the fermentation temperature of 35-37 ℃, the pressure of 0.04-0.06 MPa and the rotation speed of 60-90 r/min; stopping fermentation when the pH value is reduced to 3.0-3.5 to obtain fermentation liquor, and freezing or coating and drying the fermentation liquor to obtain the microecological bactericide.

2. The method for producing a microecological microbial agent according to claim 1, wherein the seed solution of Saccharomyces boulardii is obtained by inoculating Saccharomyces boulardii to a YPD liquid medium and culturing at 28 ℃ for 40 hours; the Bifidobacterium animalis seed solution is prepared by inoculating Bifidobacterium animalis into TPY liquid culture medium, and anaerobically culturing at 37 deg.C for 18 hr to obtain Bifidobacterium animalis seed solution; the preparation method of the lactobacillus casei seed liquid comprises the steps of inoculating lactobacillus casei into an MRS liquid culture medium, and culturing for 24 hours at 37 ℃ to obtain the lactobacillus casei seed liquid.

3. The preparation method of the microecological microbial inoculum according to claim 1, wherein the mixed culture medium comprises the following components in parts by weight: soybean peptone 1.5-2.0 parts, corn steep liquor dry powder 0.2-0.4 part, glucose 2.0-3.0 parts, soluble starch 0.1-0.2 part, K₂HPO₄·3H₂0.1-0.2 part of O and MgSO₄0.01 to 0.03 part, 0.1 to 0.2 part of L-cysteine and 800.1 to 0.2 part of Tween.

4. A microecological agent produced by the production method according to any one of claims 1 to 3.

5. The microecological agent according to claim 4, wherein the microecological agent comprises live bacteria of Bifidobacterium animalis, Lactobacillus casei and Saccharomyces boulardii, and metabolites of these three bacteria; preferably, the number of Bifidobacterium animalis is more than or equal to 1.0 × 10⁸CFU/g, the number of lactobacillus casei is more than or equal to 2.0 multiplied by 10⁸CFU/g, the number of bacteria of Saccharomyces boulardii is not less than 1.0 × 10⁸CFU/g; preferably, the metabolites comprise acetic acid, lactic acid, fatty acids, amino acids, and proteases.

6. Use of the microecological agent according to claim 4 for the preparation of a medicament or feed for the treatment of digestive tract disorders in an animal and for improving the immunity of an animal.

7. A preparation method of a microecological microcapsule coating preparation is characterized by adding 0.2-0.5 part of cellulose acetate phthalate, 0.1-0.2 part of polyvinyl alcohol phthalate, 10-15 parts of skim milk powder and 2.0-4.0 parts of sodium alginate into 80.3-87.7 parts of fermentation liquor according to parts by weight, uniformly mixing, and freezing or coating and drying to form the microecological microcapsule coating preparation;

the preparation method of the fermentation liquor comprises the following steps: inoculating the Saccharomyces boulardii seed liquid to a mixed culture medium according to the inoculation amount of 2-3%, and carrying out aerobic fermentation at the fermentation temperature of 25-28 ℃, the pressure of 0.04-0.06 MPa, the rotation speed of 180-200 r/min and the ventilation volume of 200-250 m³Fermenting for 18-24 hours; adding 1-3 wt% of sugar into the fermentation system, and adjusting the pH value to 7.0; inoculating the bifidobacterium animalis seed liquid and the lactobacillus casei seed liquid to a fermentation system according to the inoculation amounts of 1-2% and 2-3% respectively, and carrying out anaerobic fermentation at the fermentation temperature of 35-37 ℃, the pressure of 0.04-0.06 MPa and the rotation speed of 60-90 r/min; and stopping fermentation when the pH value is reduced to 3.0-3.5 to obtain fermentation liquor.

8. A coated microecological microcapsule prepared by the process according to claim 7.

9. Root of herbaceous plantThe microecological microcapsule coating formulation according to claim 8, wherein the microecological microcapsule coating formulation comprises live bacteria of Bifidobacterium animalis, Lactobacillus casei and Saccharomyces boulardii, and metabolites of these three bacteria; preferably, the number of Bifidobacterium animalis is more than or equal to 1.0 × 10⁸CFU/g, the number of lactobacillus casei is more than or equal to 2.0 multiplied by 10⁸CFU/g, the number of bacteria of Saccharomyces boulardii is not less than 1.0 × 10⁸CFU/g; preferably, the metabolites comprise acetic acid, lactic acid, fatty acids, amino acids, and proteases.

10. Use of the coated formulation of microecological microcapsules of claim 8 for the preparation of a medicament or feed for the treatment of digestive tract disorders in animals and for improving the immunity of animals.

Technical Field

The invention belongs to the technical field of feed additives, and particularly relates to a micro-ecological microcapsule coating preparation and a preparation method thereof.

Background

The microbial ecosystem in the animal body is composed of countless microbes such as bacteria, viruses and fungi, various beneficial bacteria and harmful bacteria play different roles in the animal body in a delicate balanced state, and the healthy and balanced in-vivo microbial ecosystem is beneficial to the metabolism of the animal and the absorption of nutrient substances, enables the animal to glow with more life vitality and becomes healthier and more active. Therefore, proper supplementation of probiotics is the most direct way to keep the animal microecology viable.

When the gastrointestinal tract of animals is infected by pathogens, antibiotic medicines are generally used for treatment, the disease condition can be controlled within a short time at first, but the effect is more and more insignificant in recent years, on one hand, the excessive use of antibiotics destroys the balance of the intestinal tract original flora, the gastrointestinal tract is disordered, and the autoimmunity is reduced; on the other hand, multiple use of antibiotics can cause the drug resistance of bacteria to increase, and induce the generation of 'super bacteria'.

The animal feed is added with the original animal bifidobacterium and lactobacillus casei, can be rapidly planted on the surface of intestinal mucosa, assists in establishing new flora balance to form a mucosal flora barrier, reduces the attachment and reproduction of pathogens by secreting bacteriostat, reducing the pH of the environment and resisting planting, and reduces the probability of infection; meanwhile, the generation of antibodies such as intestinal mucosa SIgA and the like is increased through mucosal immunity, and the humoral immunity of animals is improved. The saccharomyces boulardii is added, so that harmful bacteria such as escherichia coli and salmonella can be adsorbed, digestive enzymes such as protease and lipase can be secreted, digestion and absorption of food are promoted, and the animal is assisted to recover health.

At present, bifidobacterium animalis, lactobacillus casei and saccharomyces boulardii are all applied to animals, but have the defects in several aspects in the whole view: firstly, there is no synergistic effect between bacteria, the growth and reproduction speed is slow after entering gastrointestinal tract, the metabolite production is less, and the effect is not obvious; secondly, the three kinds of bacteria are separately fermented and produced, the bacterial quantity is not high, and the three kinds of bacteria are compounded into a product, so that the manufacturing cost is high, and the popularization is not facilitated; thirdly, after long-term storage at normal temperature, feed granulation and digestion of gastric juice, pancreatic juice, bile and the like, the loss of the three bacteria is more, and the three bacteria cannot continuously and effectively play a role. Therefore, how to increase the bacterial quantity and the metabolic products through the synergistic mixed fermentation and improve the tolerance of the products to the high-temperature environment and digestive juice become key problems to be solved urgently.

Disclosure of Invention

In order to overcome the defects in the prior art, the invention provides a preparation method of a microecological bactericide, which is prepared by mixing and fermenting animal bifidobacteria, lactobacillus casei and saccharomyces boulardii to form the microecological bactericide with high bacteria content and high metabolite content, and reduces the production cost. The technical scheme of the invention is as follows:

a preparation method of a microecological bactericide comprises the following steps:

(1) aerobic fermentation

Inoculating the Saccharomyces boulardii seed liquid to a mixed culture medium according to the inoculation amount of 2-3%, and carrying out aerobic fermentation at the fermentation temperature of 25-28 ℃, the pressure of 0.04-0.06 MPa, the rotation speed of 180-200 r/min and the ventilation volume of 200-250 m³And (h), defoaming at regular time, and fermenting for 18-24 h.

(2) Anaerobic fermentation

Adding 1-3 wt% of sugar into the fermentation system in the step (1), and adjusting the pH value to 7.0; inoculating the bifidobacterium animalis seed liquid and the lactobacillus casei seed liquid to a fermentation system according to the inoculation amounts of 1-2% and 2-3% respectively, and carrying out anaerobic fermentation at the fermentation temperature of 35-37 ℃, the pressure of 0.04-0.06 MPa and the rotation speed of 60-90 r/min; stopping fermentation when the pH value is reduced to 3.0-3.5 to obtain fermentation liquor, and freezing or coating and drying the fermentation liquor to obtain the microecological bactericide.

In the preparation method of the microecological bactericide, the preparation method of each seed solution comprises the following steps: inoculating bifidobacterium animalis into a TPY liquid culture medium, and carrying out anaerobic culture for 18h at the temperature of 37 ℃ to obtain a bifidobacterium animalis seed solution; inoculating lactobacillus casei into an MRS liquid culture medium, and culturing for 24h at 37 ℃ to obtain lactobacillus casei seed liquid; inoculating Saccharomyces boulardii into YPD liquid culture medium, and culturing at 28 deg.C for 40 hr to obtain Saccharomyces boulardii seed solution. Before the micro-ecological bacteria preparation is prepared by inoculation, the three seed solutions are all placed in a refrigerator for refrigeration and standby.

In the preparation method of the microecological bactericide, the preparation method of the mixed culture medium comprises the following steps: soybean peptone 1.5-2.0 weight parts, corn steep liquor dry powder 0.2-0.4 weight parts, glucose 2.0-3.0 weight parts, soluble starch 0.1-0.2 weight parts,

K₂HPO₄·3H₂0.1 to 0.2 part by weight of O and MgSO₄0.01 to 0.03 weight part, 0.1 to 0.2 weight part of L-cysteine and 800.1 to 0.2 weight part of tween, adjusting the pH value to 6.2 to 6.4, performing steam sterilization under the condition of 0.1MPa for 15min, and naturally cooling to 30 ℃ for later use.

The microecological bactericide contains Bifidobacterium animalis (number of bacteria ≧ 1.0 × 10)⁸CFU/g), Lactobacillus casei (number of bacteria ≧ 2.0 × 10)⁸CFU/g) and Saccharomyces boulardii (number of bacteria ≧ 1.0X 10)⁸CFU/g), and metabolites of these three bacteria. Wherein, the metabolites mainly comprise acetic acid, lactic acid, fatty acid, amino acid, protease and the like.

The microecological bactericide prepared by the preparation method has a good effect of improving the digestive tract diseases of animals, including dyspepsia, diarrhea, constipation and the like, and the immunity of the animals.

In order to adapt to the digestion of animals, in the preparation method of the microecological bactericide, after the fermentation liquor is obtained, the fermentation liquor can be prepared into a microecological microcapsule coating preparation so as to improve the tolerance of animal bifidobacterium, lactobacillus casei and saccharomyces boulardii to the digestion liquor of the animals and reduce the loss of beneficial strains, thereby enabling the strains to effectively play roles. The preparation method of the micro-ecological microcapsule coating preparation comprises the following steps: adding 0.2-0.5 part by weight of Cellulose Acetate Phthalate (CAP), 0.1-0.2 part by weight of polyvinyl alcohol phthalate (PVAP), 10-15 parts by weight of skimmed milk powder and 2.0-4.0 parts by weight of sodium alginate into 80.3-87.7 parts by weight of fermentation liquor, uniformly mixing, freezing or coating and drying to form the micro-ecological microcapsule coating preparation.

The microecological microcapsule coating preparation contains Bifidobacterium animalis (number of bacteria ≧ 1.0 × 10)⁸CFU/g), Lactobacillus casei (number of bacteria ≧ 2.0 × 10)⁸CFU/g) and Saccharomyces boulardii (number of bacteria ≧ 1.0X 10)⁸CFU/g), and metabolites of these three bacteria. Wherein, the generationThe metabolites are mainly acetic acid, lactic acid, fatty acid, amino acid, protease, etc.

The micro-ecological microcapsule coating preparation prepared by the preparation method has better improvement effect on animal digestive tract diseases including dyspepsia, diarrhea, constipation and the like and animal immunity.

The invention has the beneficial effects that:

the three strains of bifidobacterium animalis, lactobacillus casei and saccharomyces boulardii are mixed and fermented, so that the synergy among the strains is realized, the growth and reproduction speed is high after the strains enter the gastrointestinal tract, the generation amount of metabolites is large, and the use effect is obviously improved.

Compared with the single fermentation production of three kinds of bacteria, the mixed fermentation has high bacteria amount and relatively low production cost, and is beneficial to clinical popularization and use.

The invention also coats the microbial inoculum after mixed fermentation by a microcapsule coating technology, effectively avoids the loss of the microbial inoculum after feed granulation, digestion of gastric juice, pancreatic juice, bile and the like and long-term storage at normal temperature, plays the effective role of the microbial inoculum and prolongs the shelf life.

Drawings

FIG. 1 is a diagram of a test chicken house;

FIG. 2 is an egg laid by a chicken after eating the micro-ecological microcapsule coated preparation of the present invention;

FIG. 3 is a photograph of a boar in column 4 prior to consuming the coated microecological microcapsule of the present invention;

fig. 4 is a picture of a sow in the 8 th sow before consuming the micro-ecological microcapsule coating preparation of the present invention;

FIG. 5 is a photograph of a boar in column 4 after consuming the coated preparation of the micro-ecological microcapsules of the present invention;

fig. 6 is a picture of a sow in the 8 th sow after eating the micro-ecological microcapsule coating preparation of the invention;

FIG. 7 is a graph of diarrhea rate in piglets at column 4 and column 8 as a function of days.

Detailed Description

Terms used in the present invention have generally meanings as commonly understood by one of ordinary skill in the art, unless otherwise specified.

Comparison of the number of bacteria and metabolites between the Mixed fermentation and the Individual fermentation

(1) Fermentation of bifidobacterium animalis: inoculating the bifidobacterium animalis seed liquid to a mixed culture medium according to the inoculation amount of 1 percent, carrying out anaerobic fermentation, adjusting the fermentation temperature to 37 ℃, controlling the tank pressure to be 0.05MPa and the rotating speed to be 80r/min, and stopping fermentation when the pH value is reduced to 3.5.

(2) Fermentation of lactobacillus casei: inoculating the lactobacillus casei seed liquid to a mixed culture medium according to the inoculation amount of 2 percent, carrying out anaerobic fermentation, adjusting the fermentation temperature to 37 ℃, the tank pressure to 0.05MPa, the rotating speed to 80r/min, and stopping fermentation when the pH value is reduced to 3.5.

(3) Fermentation of Saccharomyces boulardii: inoculating the Saccharomyces boulardii seed liquid to the mixed culture medium according to the inoculation amount of 2%, and performing aerobic fermentation at 28 deg.C, the tank pressure of 0.05MPa, the rotation speed of 200r/min, and the air flow of 250m³And/h, defoaming at regular time and fermenting for 18 h.

(4) Mixed fermentation of bifidobacterium animalis, lactobacillus casei and saccharomyces boulardii: inoculating the Saccharomyces boulardii seed liquid to the mixed culture medium according to the inoculation amount of 2%, and performing aerobic fermentation at 28 deg.C, the tank pressure of 0.05MPa, the rotation speed of 200r/min, and the air flow of 250m³And/h, defoaming at regular time and fermenting for 18 h. Adding 2 weight percent of glucose into the fermentation system, and adjusting the pH value to 7.0; inoculating Bifidobacterium animalis seed liquid and Lactobacillus casei seed liquid to a fermentation system according to the inoculation amount of 1% and 2%, performing anaerobic fermentation at 37 deg.C under 0.05MPa at 80r/min, and stopping fermentation when pH value is reduced to 3.5.

In the fermentation process, the preparation method of each seed liquid comprises the following steps: inoculating bifidobacterium animalis into a TPY liquid culture medium, and carrying out anaerobic culture for 18h at the temperature of 37 ℃ to obtain a bifidobacterium animalis seed solution; inoculating lactobacillus casei into an MRS liquid culture medium, and culturing for 24h at 37 ℃ to obtain lactobacillus casei seed liquid; inoculating Saccharomyces boulardii into YPD liquid culture medium, and culturing at 28 deg.C for 40 hr to obtain Saccharomyces boulardii seed solution.

In the above fermentation process, the mixed culture medium is preparedThe preparation method comprises the following steps: 2.0 parts of soybean peptone, 0.4 part of corn steep liquor dry powder, 2.0 parts of glucose, 0.2 part of soluble starch and K₂HPO₄·3H₂O0.1 part by weight, MgSO₄0.01 weight part, 0.2 weight part of L-cysteine, 800.1 weight parts of tween and 994.99 weight parts of purified water, uniformly mixing, adjusting the pH value to 6.2, performing steam sterilization for 15min under the condition of 0.1MPa, and naturally cooling to 30 ℃ for later use.

Bifidobacterium animalis and Saccharomyces boulardii were obtained from Jiangsu Microkang and Angel Yeast, respectively, and Lactobacillus casei (strain number 1.8727) was purchased from the China center for general microbiological Collection.

The number of bacteria in the four fermented liquids was measured and counted, and the results are shown in table 1:

TABLE 1

The metabolites in the four fermentation broths were tested and the results are shown in table 2:

TABLE 2

As can be seen from the detection results in tables 1 and 2, compared with the single fermentation of the three strains, the final bacteria number and the main metabolites are obviously improved after the mixed fermentation, which indicates that the three strains can mutually promote the growth and the propagation in the mixed fermentation process.

The present invention will be described in further detail with reference to the following data in conjunction with specific examples. The following examples are intended to illustrate the invention and are not intended to limit the scope of the invention in any way.

Example 1

Preparing a microecological bactericide:

(1) preparation of seed liquid

Inoculating bifidobacterium animalis into a TPY liquid culture medium, and carrying out anaerobic culture for 18h at the temperature of 37 ℃ to obtain a bifidobacterium animalis seed solution; inoculating lactobacillus casei into an MRS liquid culture medium, and culturing for 24h at 37 ℃ to obtain lactobacillus casei seed liquid; inoculating Saccharomyces boulardii into YPD liquid culture medium, and culturing at 28 deg.C for 40 hr to obtain Saccharomyces boulardii seed solution. And (4) placing the seed liquid of the three kinds of bacteria in a refrigerator for refrigeration for standby.

(2) Preparation of Mixed Medium

2.0 parts of soybean peptone, 0.4 part of corn steep liquor dry powder, 2.0 parts of glucose, 0.2 part of soluble starch and K₂HPO₄·3H₂O0.1 part by weight, MgSO₄0.01 weight part, 0.2 weight part of L-cysteine, 800.1 weight parts of tween and 994.99 weight parts of purified water, uniformly mixing, adjusting the pH value to 6.2, performing steam sterilization for 15min under the condition of 0.1MPa, and naturally cooling to 30 ℃ for later use.

(3) Aerobic fermentation

Inoculating the Saccharomyces boulardii seed liquid to the mixed culture medium according to the inoculation amount of 2%, and performing aerobic fermentation at 28 deg.C, the tank pressure of 0.05MPa, the rotation speed of 200r/min, and the air flow of 250m³And/h, defoaming at regular time and fermenting for 18 h.

(4) Anaerobic fermentation

Adding 2 weight percent of glucose into the fermentation system, and adjusting the pH value to 7.0; inoculating bifidobacterium animalis seed liquid and lactobacillus casei seed liquid to a fermentation system according to the inoculation amounts of 1% and 2% respectively, and carrying out anaerobic fermentation, wherein the fermentation temperature is adjusted to 37 ℃, the tank pressure is 0.05MPa, and the rotating speed is 80 r/min; stopping fermentation when pH value is reduced to 3.5 to obtain fermentation liquor.

(5) Freezing or coating drying

And (4) freezing or coating and drying the fermentation liquor to obtain the microecological bactericide.

Example 2

Preparation of a micro-ecological microcapsule coating preparation:

adding 0.2 part by weight of Cellulose Acetate Phthalate (CAP), 0.2 part by weight of polyvinyl alcohol phthalate (PVAP), 15 parts by weight of skim milk powder and 4.0 parts by weight of sodium alginate into 80.6 parts by weight of the fermentation liquid prepared in the step (5) of the example 1, uniformly mixing, and freezing or coating and drying to form the micro-ecological microcapsule coating preparation.

Example 3 tolerance test for digestive juices

The microecological microcapsule-coated preparation prepared in example 2 was tested for tolerance to digestive juices, using the microecological agent (example 1) as a control and the survival rate of three bacteria as a tolerance indicator.

The digestive juice is replaced by artificial gastric juice and artificial intestinal juice.

Artificial gastric juice: taking 16.4mL of dilute hydrochloric acid, adding about 800mL of water and 10g of pepsin (3000NFU/g, Solebao company), stirring uniformly, adding water and fixing the volume to 1000 mL.

Artificial intestinal juice: adding 500mL of water into 6.8g of monopotassium phosphate, and adjusting the pH to 6.8 by using 0.4% sodium hydroxide solution; in addition, 10g of pancreatin (250NFU/g, Solebao Co.) was dissolved in a suitable amount of water, and the two solutions were mixed and then added with water to a constant volume of 1000 mL.

Adding the bacterial powder sample into the artificial gastric juice and the intestinal juice according to the concentration of 1%, shaking for 2 hours and 4 hours respectively at the temperature of 37 ℃ by a shaking table at 80r/min, detecting the bacterial count before and after treatment, and calculating the survival rate.

The survival rate of the bacteria is equal to the number of bacteria after treatment (artificial gastric juice or artificial intestinal juice)/the number of bacteria before treatment multiplied by 100 percent

The test results are shown in table 3:

TABLE 3

As can be seen from Table 3, if the mixed fermentation is directly spray-dried, the obtained microbial inoculum is digested by artificial gastric juice and artificial intestinal juice, and the loss of the number of bacteria is large; the survival rate of the coated micro-capsules is over 75 percent, so that the bacteria can smoothly reach the intestinal tract to play a role.

Example 4 Effect of Normal temperature storage on the viability of bacteria

The microbial viability of the coated preparation of the micro-ecological microcapsule prepared in example 2 under normal temperature storage (1 month, 3 months, 6 months, 12 months) was tested, using the micro-ecological microbial agent (example 1) as a control and the viability of three bacteria as a detection index.

The test results are shown in table 4:

TABLE 4

As can be seen from table 4, if the mixed fermentation is directly spray-dried, the obtained microbial inoculum still maintains a high survival rate after being placed at normal temperature for 3 months, but the number of bifidobacterium animalis and lactobacillus casei is reduced to below 60%, and the microbial inoculum is lost after being stored for 12 months and cannot effectively play a role; after the microcapsule coating, the survival rate of the bacteria is still over 70 percent after the bacteria are stored for 12 months, and the using effect of the product is ensured.

Example 5

Mouse safety test: 30 Kunming mice (male and female halves) with the age of 4 weeks are randomly divided into a control group, a low-dose group and a high-dose group (10 mice respectively), the mice are respectively perfused with physiological saline, a low-dose microecological microcapsule coating preparation (0.001g/mL prepared by the physiological saline) and a high-dose microecological microcapsule coating preparation (0.1g/mL prepared by the physiological saline), 1mL is carried out every time, 1 time is carried out every day, the phenomena of death and diarrhea are observed every day, the weight is weighed after the intragastric administration is carried out for 14 days, the visceral diseases are observed in an anatomical mode, the liver, the spleen and the kidney are weighed, and the visceral organ index is calculated.

Organ index is weight/body weight of the organ (liver, spleen or kidney) x 100%.

The index of the mouse organs 14 days after gavage is shown in table 5.

TABLE 5

Through experimental observation, the mice do not die and have obvious diarrhea phenomenon in the experimental period. After 14 days of gastric lavage, the stomach and the intestinal tract were dissected and observed, and no obvious lesions were found. As can be seen from Table 5, compared with the control group, the indexes of the liver, spleen and kidney of the low-dose and high-dose microecological microcapsule coating preparations in the gavage group have no obvious change, which indicates that the microecological microcapsule coating preparation prepared by the mixed fermentation of the three bacteria has no toxic or side effect.

Clinical case testing and analysis

(1) Case one

Variety: labrador retriever

Age: 2 years and 6 months

Sex: male sex

Symptoms are: diarrhea for 1 more months, orbit subsidence, stink excrement, no formation, occasional blood, borborborygmus, weight loss of about 0.9kg, inappetence, and mental depression.

Administration: when norfloxacin is used for treating for 1 week, the effect is not ideal, and after the norfloxacin is used, the compound microecological preparation (bifidobacterium, enterococcus faecalis and yeast) is used, the borborygmus is not obvious after the norfloxacin is used, the feces are not stinking, and the diarrhea phenomenon still exists.

Prescription: the microecological microcapsule coating preparation is used for 1 time a day, 10g each time and 5 days continuously.

As a result: the feces begin to form the shape the next day after use, the water feces and bloody feces disappear, the normal food intake and drinking are recovered the fourth day, the mental state is recovered to normal the fifth day, and the body weight is basically recovered to the previous level after half a month of return visit.

(2) Case two

Variety: teddy dog

Age: 1 year 4 months

Sex: male sex

Symptoms are: the sick dog has no appetite, depressed spirit, dysphagia, spitting mucus food, fishy smell, irregular cough, esophagus obstruction detected by hospital, food reflux, and megaesophagus disease.

Administration: the cisapride is used for promoting the peristalsis of the digestive tract by 0.25mg/kg, the omeprazole is used for reducing the gastric acid secretion by 1.0mg/kg, and then the compound microecologics (bifidobacterium, lactobacillus and streptococcus thermophilus) are used, so that no obvious effect is produced after the compound microecologics are used.

Prescription: the micro-ecological microcapsule coating preparation is mixed into warm water with more than 30 ℃ for pouring, 2 times a day, 5g each time and 5 days for continuous use.

As a result: the symptoms of dysphagia and cough are relieved on the second day after the use, food is not spitted on the third day, and the appetite is recovered to be normal.

(3) Case three

Variety: hailan brown egg chicken

Age: 1 year 6 months

Scale: 7000 only

Symptoms are: the feed has obvious diarrhea and peritonitis, the diarrhea rate is about 25 percent, the fatality rate is 1.2 percent, the number of malformed eggs and preserved sand eggs is large, the egg yield is reduced, and on average, each chicken lays 60g of eggs every day. The test chicken house is shown in figure 1.

Prescription: the micro-ecological microcapsule coating preparation of the invention is mixed with feed according to the dosage of 0.1 percent and is used for 5 days continuously.

As a result: after the feed is used, the diarrhea rate is reduced to 5% on the third day, death does not occur on the fourth day, each chicken can lay eggs by 63g every day on the sixth day, deformed eggs and preserved sand eggs disappear, and the economic benefit is obviously improved. The eggs laid by the chickens fed with the micro-ecological microcapsule coating preparation are shown in figure 2.

(4) Case four

Animals: about 33 piglets at 35 days of age, 15 boars at column 4 (as shown in fig. 3) and 18 sows at column 8 (as shown in fig. 4).

Indexes are as follows: diarrhea rate and hair color were scored.

Symptoms are: coarse and disorderly hair, protruding scapula and spine, diarrhea and poor appetite.

Prescription: the micro-ecological microcapsule coating preparation of the invention is mixed with feed according to the dosage of 0.1 percent and is used for 14 days continuously.

As a result: the diarrhea rate and the hair color score and the condition of the pigs are shown in fig. 5 and fig. 6, and tables 6 and 7, wherein fig. 5 shows the living state of the 4 th boar after eating the microecological microcapsule-coated preparation of the present invention, fig. 6 shows the living state of the 8 th sow after eating the microecological microcapsule-coated preparation of the present invention, fig. 7 shows the diarrhea rate of the piglets as a function of days, and tables 6 show the hair color scores of the piglets before and after the test.

TABLE 6

As can be seen from fig. 7, the diarrhea rate is continuously reduced from the severe period to the end of complete recovery, which shows that the micro-ecological microcapsule coating preparation of the invention has the function of relieving and eliminating the diarrhea of piglets. As can be seen from fig. 3 to 6 and table 6, after the test for 1 week and 2 weeks, the hair color of 4-column piglets is respectively improved by 10.5% and 31.4%, and the qualification rate is respectively improved by 20.0% and 60.0%; the hair color of 8 piglets is respectively improved by 57.7 percent and 60.3 percent, the qualification rate is respectively improved by 55.6 percent and 61.1 percent, which shows that the microecological microcapsule coating preparation has good health care and rescue effects on the piglets.

The foregoing is directed to preferred embodiments of the present invention, other and further embodiments of the invention may be devised without departing from the basic scope thereof, and the scope thereof is determined by the claims that follow. However, any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present invention are within the protection scope of the technical solution of the present invention.