CN114190486B

CN114190486B - Microbial fermentation feed for preventing and treating constipation of sows and sow breeding method

Info

Publication number: CN114190486B
Application number: CN202111538894.6A
Authority: CN
Inventors: 李建涛; 刘显军; 李海东; 程奇
Original assignee: Shenyang Agricultural University
Current assignee: Shenyang Agricultural University
Priority date: 2018-08-08
Filing date: 2018-08-08
Publication date: 2024-08-13
Anticipated expiration: 2038-08-08
Also published as: CN114190486A; CN108813106A

Abstract

The invention relates to a microbial fermentation feed for preventing and treating constipation of sows and a sow breeding method, wherein the feed comprises the following raw material components in parts by weight: 28 to 31 parts of corn, 25 to 27 parts of bean pulp, 4 to 6 parts of DDGSs, 9 to 12 parts of corn gluten feed, 9 to 11 parts of corn germ meal, 17 to 19 parts of rice bran meal, 1.5 to 2.5 parts of stone powder, 0.3 to 1 part of senna leaf, 0.1 to 0.3 part of aerobic fermentation agent and 0.1 to 0.3 part of anaerobic fermentation agent. The feed product can meet the nutrition types required by sows, the carbon nitrogen ratio, pH, moisture, fermentable carbohydrate, osmotic pressure and the like required by microbial growth, and the proper microorganisms are screened to ferment the compound feed aiming at the physiological characteristics of constipation sows, so that the compound feed has the effects of improving the feed raw material components and the biological barrier of the intestinal tracts of the constipation sows, preventing and treating the constipation of the sows and improving the reproductive performance of the sows.

Description

Microbial fermentation feed for preventing and treating constipation of sows and sow breeding method

The application relates to a microbial fermented feed for preventing and treating constipation of sows and a use method thereof, which are divisionally applied for the application of 2018, 08 and 08, 201810894039.0.

Technical Field

The invention relates to the technical field of biology, in particular to a microbial fermented feed for preventing and treating constipation of sows and a sow breeding method, namely a preparation method of the fermented feed for preventing and treating constipation of sows by adopting microbial synergistic dry feed mixed feed and a sow breeding method.

Background

The sow is the root and the core of pig farm development, and the pig farm benefit is left and right. Therefore, the quality of sow feeding determines the future development of a pig farm. However, the occurrence of some diseases in the sow raising process can affect the performance of sow production. Sow constipation is a relatively easily occurring disease, especially in the late gestation period and before and after birth, and sometimes the incidence rate can be up to 60%. Constipation of sow can cause imbalance of metabolism of organism, endocrine disturbance, depression of the spirit, inappetence, complete waste of appetite in serious cases, difficult defecation, abdominal pain and the like. After the pregnant sow is sick, the excrement formed by metabolism of the organism cannot be discharged for a long time, so that the toxin in the body is increased, the toxin invades the embryo, the development of the embryo is influenced, the abortion or delivery time is increased, even the dystocia phenomenon occurs, or malformation or weaned piglets are produced, and even the stillbirth is produced. Infection during lactation will affect lactation, and lactation disorder occurs, which affects survival and production performance of piglets, and seriously damages economic benefit of pig industry.

The sow's availability of adequate and balanced nutrition is the basis for its own reproductive potential. Limited feeding, lack of exercise, lack of green feed, lack of drinking water and the like are all causes of constipation of sows, and severely restrict the development of reproductive potential. However, the traditional way of preventing and treating constipation of sows has a plurality of defects: increasing the fiber content in the diet reduces the effective level of the diet, which contradicts the high-concentration nutritional requirements of modern genotype sows; the long-term feeding of mineral salt laxatives can destroy electrolyte balance in the body, and cause metabolic disorder; the feeding of the green feed is difficult to implement in large scale pig farms.

From the 50 th century of the 20 th century, the fermented feed is fed into the field of vision of people, researchers propose to ferment the nondigestible coarse feed into the coarse feed which can be digested better for pigs, and experimental researches on piglets, sows and fattening pigs show that the fermented feed has a good feeding effect. The microbial fermented feed is a novel biological feed obtained by carrying out aerobic or anaerobic fermentation on one or more feed raw materials or compound feed by using microorganisms under artificial control conditions, eliminating anti-nutritional factors, reducing the crude fiber content of the fermented raw materials by fermentation, degrading substrates in the fermentation process, improving the utilization efficiency of nutrients, and enriching a large amount of active probiotics and functional primary/secondary metabolites. The microbial fermented feed has animal nutrition and intestinal microecology balance, and has the function of immunity and health care. The fermented feed product has sour flavor, improves the palatability of the feed, and stimulates the feed intake of pigs.

Although the fermented feed achieves good application effect, in actual production, strain sources are unknown, so that strain degradation is serious and physiological activity is reduced. Meanwhile, the strains are monotonous, and the cooperation of different strains is lacking, so that the fermentation effect is influenced.

At present, the consumer has a higher call for healthy food, and the consumer has an increased importance for green healthy cultivation. Therefore, research on novel safe, green and efficient microbial fermentation feed products with the effects of promoting animal growth, improving health conditions, being beneficial to environment and the like becomes a new way for development of animal husbandry.

Disclosure of Invention

In order to overcome the defects of the traditional fermentation technology for preventing and treating constipation of sows, the invention aims to provide the microbial fermentation feed and the cultivation method for preventing and treating constipation of sows, and proper feed raw materials are selected to prepare dry mixed feed, so that the feed product meets the nutrition types required by sows and the carbon nitrogen ratio, pH, moisture, fermentable carbohydrates, osmotic pressure and the like required by microbial growth, and proper microorganisms are selected to ferment the compound feed aiming at the physiological characteristics of constipation sows, so that the compound feed has the functions of improving feed raw material components and biological barriers of intestinal tracts of constipation sows, and finally achieves the purposes of preventing and treating constipation of sows and improving reproductive performance of sows.

The invention aims at realizing the following technical scheme:

The invention provides a microbial fermentation feed for preventing and treating constipation of sows, which is technically characterized by comprising the following components in parts by weight: the microbial fermentation feed for preventing and treating constipation of sows comprises the following raw materials in parts by weight: 28 to 31 parts of corn, 25 to 27 parts of bean pulp, 4 to 6 parts of DDGS, 9 to 12 parts of corn gluten feed, 9 to 11 parts of corn germ meal, 17 to 19 parts of rice bran meal, 1.5 to 2.5 parts of stone powder, 0.3 to 1 part of senna leaf, 0.1 to 0.3 part of aerobic fermentation agent and 0.1 to 0.3 part of anaerobic fermentation agent;

the aerobic fermentation agent comprises bacillus subtilis, agrocybe cylindracea fungi and straw mushroom fungi, and the weight ratio of the bacillus subtilis to the agrocybe cylindracea fungi is 1:1:1, a step of;

The Agrocybe cylindracea B002 fungus has a preservation unit of China general microbiological culture Collection center, a preservation address of North Star, a China academy of sciences of China, west Lu 1, no. 3, the Korean area of Beijing, a preservation time of 2018, 7 months and 12 days, a preservation number of CGMCC No.16085, and a classification name of Agrocybe cylindracea Agrocybe aegerita;

the agrocybe cylindracea fungus microbiology is characterized in that spores germinate on PDA culture medium at 23-28 ℃ for 24 hours, and tiny hyphae can be seen by naked eyes after 48 hours. Fungus ring is white, membranous and superior. Oval to elliptical spores. Temperature range for hypha growth: the proper temperature is between 10 and 35 ℃, the proper temperature is between 22 and 26 ℃, and the proper pH for growth is: 5 to 7.5;

The collection unit of the straw mushroom B004 fungus is China general microbiological culture Collection center, the collection address is China national academy of sciences of China sciences of China No.3 of West road 1, korean area North Star of Beijing, the collection time is 2018, 7 months and 12 days, the collection number is CGMCC No.16086, and the classification and naming are straw mushrooms Volvariella volvacea;

The fungus microbiology features colorless and transparent mycelium, the length of straw mushroom cell is 46-400 microns, the width is 6-18 microns, the cell is separated into multicellular mycelium by membrane, the mycelium is continuously branched and spread, the mycelium is interwoven with each other to form loose net mycelium, the culture temperature is 10-40 ℃, the proper temperature is 28-33 ℃, the culture pH is 4.0-10.3, and the proper pH at the stage of mycelium and fruiting body is 4.7-6.5. The relative humidity of air is 75-85%.

The anaerobic fermentation agent comprises enterococcus faecalis and Saccharomyces cerevisiae, and the dosage ratio of the enterococcus faecalis to the Saccharomyces cerevisiae is 2:1.

Preferably, the microbial fermented feed for preventing and treating constipation of sows comprises the following raw material components in parts by weight: 30 parts of corn, 25 parts of bean pulp, 5 parts of DDGS, 10 parts of corn gluten feed, 9.4 parts of corn germ meal, 18 parts of rice bran meal, 2 parts of stone powder, 0.6 part of senna leaf, 0.2 part of aerobic fermentation agent and 0.2 part of anaerobic fermentation agent.

The invention provides a specific preparation method of microbial fermented feed for preventing and treating constipation of sows, which comprises the following steps:

1) Weighing qualified corns, bean pulp, rice bran meal, DDGS, corn germ meal, corn husks, stone powder and senna leaves according to the parts by weight, crushing to obtain raw materials with the particle size of 2.0mm, putting the raw materials into a stirring tank, stirring at the stirring speed of 30-50r/m, and mixing for 10-20 minutes to obtain a dry material with the variation coefficient of less than 7%;

2) Preparation of aerobic fermentation agent

Preparation of bacillus subtilis: inoculating activated bacillus subtilis into a 100mL conical flask filled with 50mL of common broth culture medium in a sterile laboratory, and culturing for 8-14 h at 35-38 ℃ to obtain liquid bacillus bacterial liquid, wherein the number of viable bacteria in the bacterial liquid is more than or equal to 109cfu/mL;

Preparation of agrocybe cylindracea fungi: under the aseptic condition, transferring activated agrocybe cylindracea strain to PAD slant culture medium, and placing the agrocybe cylindracea strain in an incubator at 20-27 ℃ for culture, wherein the strain culture condition is as follows: intermittently ventilating the mycelium growing chamber for 1-2 times per day, wherein the relative humidity is preferably 65% -70%; the mycelium grows up the inclined plane to be the agrocybe cylindracea fungus strain; the pH value of the PAD slant culture medium is 5.5-6.5;

Preparation of straw mushroom fungi: inoculating activated straw mushroom strains to a glass test tube PAD culture medium under a sterile condition, culturing at 10-40 ℃ for 3-5 days until mycelia fully grow on the culture medium, and storing at 15-20 ℃;

the bacillus subtilis, agrocybe cylindracea fungi and straw mushroom fungi are mixed according to the weight ratio of 1:1:1, mixing;

3) Diluting the aerobic fermentation agent 100-250 times into clean water, stirring uniformly, pumping the aerobic fermentation agent into the raw material stirring tank of the step 1), mixing for 5-10 minutes to uniformly distribute bacterial liquid and raw material components, preparing a wet material I, putting the wet material I into a feed bag, warehousing and standing for ventilation fermentation, wherein the warehouse environment temperature is 23-28 ℃, and the fermentation time is as follows: fermenting for 3-5 days in winter and fermenting for 2-3 days in other seasons;

4) Preparation of anaerobic fermentation agent

Preparation of enterococcus faecalis: inoculating the activated enterococcus faecalis into an MRS culture medium under the aseptic condition, and culturing for 13-16 hours at the temperature of 35-37 ℃ in an anaerobic incubator, wherein the number of viable bacteria in bacterial liquid is more than or equal to 10 ¹⁰ cfu/mL;

Preparation of Saccharomyces cerevisiae: inoculating activated saccharomyces cerevisiae into PAD liquid culture medium under aseptic condition, culturing for 10-16 h at 28-32 ℃ in anaerobic incubator, wherein the number of viable bacteria in bacterial liquid is more than or equal to 10 ¹⁰ cfu/mL;

The enterococcus faecalis and the saccharomyces cerevisiae are mixed according to the weight ratio of 2: 1.

5) Diluting the anaerobic fermentation agent 30-100 times into clean water, stirring uniformly, putting the fermented wet material I into a stirring tank, stirring at a stirring speed of 30-40 r/m, mixing for 10-20 minutes, pumping the anaerobic fermentation agent into the fermented wet material I, mixing the anaerobic fermentation agent bacterial liquid and the fermented wet material I uniformly for 5-10 minutes again to obtain a wet material II, putting the wet material II into a breathing bag with an exhaust hole, sealing a bag opening, warehousing, standing and fermenting, stacking layers less than or equal to 5, and keeping the ambient temperature of a storehouse at more than or equal to 25 ℃ for fermentation time: fermenting for 2-3 days in winter and fermenting for 1-2 days in other seasons.

The quality technical index of the microbial fermentation feed is as follows: pH value of the fermented feed: 5.5 to 6; the moisture of the fermented feed is 30% +/-2%; the thick ferment flavor of the fermented feed can promote the appetite of sows; the hygienic index of the fermented feed meets the requirements of the hygienic standard of the feed; the microbial fermentation feed raw material and each production link cannot contain any antibiotics.

The invention provides a method for using microbial fermentation feed for preventing and treating constipation of sows, which is technically characterized by comprising the following steps:

(1) The microbial fermentation feed is uniformly mixed with the conventional feed, and the proportion of the microbial fermentation feed to the conventional feed is 1-3: 20, feeding can be performed at each stage of sow feeding;

(2) The constipation prevention is evenly mixed with the conventional batch according to the proportion of 5-10%, then is fed, the constipation prevention is evenly mixed with the conventional feed according to the proportion of 10-15%, and is continuously fed for 3-5 days, if the constipation prevention is relieved, the dosage can be reduced to be within 10%;

(3) The fermented feed can also be fed in an additional adding mode, and each sow is fed with 100-250 g of the fermented feed per day.

The invention also provides a sow breeding method, which is characterized by comprising the following steps:

the microbial fermentation feed in the technical scheme is uniformly mixed with the conventional feed, so that the feed can be fed in each stage of sow feeding; the mass ratio of the microbial fermentation feed to the conventional feed is 1-3: 20, a step of;

or when constipation is prevented, the microbial fermentation feed is fed after being evenly mixed with the conventional batch according to the mass ratio of 5-10%; when the constipation of the sow is treated, the microbial fermentation feed is uniformly mixed with the conventional feed according to the mass ratio of 10% -15%, and is continuously fed for 3-5 days, if the constipation is relieved, the dosage can be reduced to be within 10%;

Or, the fermented feed can also be fed by adopting an additional adding mode, and each sow is fed with 100-250 g of the fermented feed per day.

The invention provides a microbial fermentation feed for preventing and treating constipation of sows, which has the following remarkable advantages compared with the prior art:

1. the feed raw materials used in the invention are all conventional raw materials, the preparation process is clear, no special equipment is needed, and the feed is easy to accept and adopt.

2. According to the nutrition physiological characteristics of sows, the invention optimizes various feed raw materials to form the sow dry feed mixed feed aiming at the nutritional requirements of constipation sows on daily ration. The corn contains more than 70% of sugar substances, contains minerals such as protein, fat, calcium, phosphorus, iron and the like, also contains various vitamins such as carotene, vitamin B, amino acids and the like, and is an important energy feed in the sow dry feed mixed feed. The soybean meal is a feed prepared by moderately heating, drying and crushing residues after soybean oil extraction, the crude protein content is more than 45%, the digestibility is as high as 85% -92%, and the soybean meal is a good protein source for sows. DDGS is distillers 'grains protein feed, i.e. distillers dried grains containing solubles, which is a distillers' grains product after fermentation of ethanol from corn, wherein the content of protein, B vitamins and amino acids is increased over corn, and contains unknown growth-promoting factors generated during fermentation. Corn gluten is a byproduct of high-quality corn after starch extraction, has high crude fiber content, and is an excellent natural resource for producing high-F-value oligopeptide. Senna is the leaflet of the narrow leaf senna or the sharp leaf senna of leguminous plants, and can stimulate intestinal peristalsis through intestinal mucosa and nerves, thereby facilitating defecation. The dry mixed feed prepared by the invention not only meets the special requirements of the sow on daily ration protein, energy and crude fiber, but also is beneficial to promoting intestinal peristalsis of the sow and improving digestion function, and radically prevents constipation.

3. The invention adopts the combination of bacteria and fungi. Enterococcus faecalis and bacillus subtilis are common fermentation bacteria, saccharomyces cerevisiae is single-cell fungi, and agrocybe cylindracea and straw mushrooms are multicellular fungi.

Enterococcus faecalis is a gram-positive, hydrogen peroxide-negative coccus, which is the second only flora of E.coli in the animal's gut. Enterococcus faecalis can produce natural antibiotics, improve the immunity of the organism, secrete antibacterial substances, inhibit the growth of pathogenic bacteria such as escherichia coli and salmonella, and improve the intestinal microenvironment; meanwhile, the growth of urease-producing bacteria and putrefying bacteria in the intestinal tract can be inhibited, the content of urease and endotoxin in the intestinal tract and blood can be reduced, and the health of organisms can be facilitated. Bacillus subtilis is a gram-positive bacterium of the genus bacillus. Active substances such as subtilisin, polymyxin, nystatin, gramicidin and the like which inhibit pathogenic bacteria can be generated in the growth process of bacillus subtilis thalli; meanwhile, free oxygen rapidly consumed by the growth of thalli causes intestinal hypoxia, promotes the growth of beneficial anaerobic bacteria and indirectly inhibits the growth of other pathogenic bacteria; the bacillus subtilis can stimulate the growth and development of animal immune organs, enhance cellular immunity and humoral immunity functions and improve the immunity of organisms.

Saccharomyces cerevisiae belongs to the family of saccharomycetes, is a single-cell fungus, is in the shape of an oval or a sphere, and has simple morphology but more complex physiology. During anaerobic fermentation, saccharomyces cerevisiae sucks monosaccharide in the feed into cells, and under the action of intracellular enzyme, the monosaccharide is decomposed into carbon dioxide and alcohol. Agrocybe cylindracea is a kind of edible and medicinal fungus, and is named after being wild on the dried oil tea tree. The agrocybe cylindracea fungi is rich in nutrition, high in protein content and low in fat content. Agrocybe cylindracea mycotic Ping Ganwen has effects of clearing heat, invigorating spleen and enhancing immunity. The straw mushroom fungi has high content of essential amino acids up to 40.47-44.47%, and also contains multiple mineral elements such as phosphorus, potassium, calcium and the like.

4. The preparation method of the fermented feed utilizes a mixed bacteria aerobic and anaerobic two-step fermentation method. Aerobic bacteria bacillus subtilis, agrocybe cylindracea fungi and straw mushroom fungi are subjected to aerobic fermentation, so that the content of protein, carbohydrate, vitamins, minerals and other nutrient substances in the feed drier is improved, and a plurality of physiological active substances such as polysaccharide, polypeptide, alkaloid, terpenoid, sterol, enzyme, nucleic acid, amino acid, vitamins, plant hormone and various compounds with antibiotic effect are produced in fermentation liquid, so that nutrient-rich base material substances are provided for the later anaerobic fermentation. And then enterococcus faecalis and Saccharomyces cerevisiae are utilized for anaerobic fermentation, bacterial thalli can synthesize alpha-amylase, protease, lipase, cellulase and other enzymes, can also extract raw materials of nucleic acid derivatives, coenzyme A, cytochrome C, glutathione and various amino acids, can also synthesize various B vitamins, promote the digestion and the immunocompetence of organisms, and can reduce anti-nutritional factors and lignin in the diet, so that the feed has higher nutritional value.

In addition, the acidic products and mellow substances generated by the mixed fermentation of the multiple bacteria are beneficial to improving the acidity of the feed, improving the palatability of the feed, guaranteeing the nutrition supply of the reproductive performance of the sow, improving the intestinal microecological environment of the sow and rapidly improving the constipation of the sow.

5. The compound acidifier disclosed by the invention has the advantages of less consumption of the required leavening agent, high efficiency and only 700-900% of the proportion of the compound acidifier to the dry mixed feed: 1, and the fermented feed is mixed with the conventional feed according to the proportion of 5-15%, so that no toxic or side effect is produced on animals.

6. The fermented feed has good effect of preventing constipation, the cure rate reaches 95 percent, and constipation is basically eliminated after continuous feeding for 3 to 5 days.

Biological preservation information

Agrocybe cylindracea (Agrocybe aegerita) B002 is preserved in China general microbiological culture Collection center (China Committee for culture Collection), the preservation address is North Star, the China academy of sciences of China, no. 3, the Korean area North Star, and the preservation time is 2018, 7 months and 12 days, and the preservation number is CGMCC No.16085.

Agrocybe cylindracea (Volvariella volvacea) B004 is preserved in China general microbiological culture Collection center (China Committee for culture Collection), the preservation address is North Star, the China academy of sciences of China, no. 3, the Korean area North Star, and the preservation time is 2018, 7 months and 12 days, and the preservation number is CGMCC No.16086.

Drawings

FIG. 1 is a fecal character of a constipation sow at the beginning of the test;

Figure 2 shows the fecal character change of sows after feeding the microbial fermented feed.

Detailed Description

Example 1

(1) The microbial fermented feed for preventing and treating constipation of sows comprises the following raw materials in parts by weight: 28 to 31 parts of corn, 25 to 27 parts of bean pulp, 4 to 6 parts of DDGS, 9 to 12 parts of corn gluten feed, 9 to 11 parts of corn germ meal, 17 to 19 parts of rice bran meal, 1.5 to 2.5 parts of stone powder, 0.3 to 1 part of senna leaf, 0.1 to 0.3 part of aerobic fermentation agent and 0.1 to 0.3 part of anaerobic fermentation agent.

The aerobic fermentation agent comprises bacillus subtilis, agrocybe cylindracea fungi and straw mushroom fungi, and the weight ratio of the bacillus subtilis to the agrocybe cylindracea fungi is 1:1:1.

The agrocybe cylindracea fungi has a preservation unit of China general microbiological culture Collection center, a preservation address of Beijing being the institute of microbiology of China academy of sciences 3, west way 1, chaoyang area North Star, a preservation time of 2018, 7 months and 12 days, a preservation number of CGMCC No.16085, and is classified and named agrocybe cylindracea Agrocybe aegerita.

The agrocybe cylindracea fungus microbiology is characterized in that spores germinate on PDA culture medium at 23-28 ℃ for 24 hours, and tiny hyphae can be seen by naked eyes after 48 hours. Fungus ring is white, membranous and superior. Oval to elliptical spores. The temperature range of mycelium growth is 10-35 ℃, the proper temperature is 22-26 ℃, and the proper pH for growth is: 5 to 7.5.

The collection unit of the straw mushroom fungi is China general microbiological culture Collection center, the collection address is the China national academy of sciences of China, the Bakudzuvine is the Korea North Star, the Xiyu No.1, the Xiyu No. 3, the collection time is 2018, 7 months and 12 days, the collection number is CGMCC No.16086, and the classification and the naming are straw mushrooms Volvariella volvacea.

The fungus microbiology of straw mushroom is characterized in that hypha is colorless and transparent, the length of straw mushroom cells is 46-400 mu m, the width is 6-18 mu m, the straw mushroom cells are separated into multicellular hypha by a diaphragm, the multicellular hypha continuously branches and spreads, and the multicellular hypha interweaves with each other to form loose reticular mycelium. The culture temperature is 10-40 ℃, the proper temperature is 28-33 ℃, the culture pH is 4.0-10.3, and the proper pH at the stage of mycelium and fruiting body is 4.7-6.5. The relative humidity of air is 75-85%.

(2) The specific preparation method of the microbial fermented feed for preventing and treating constipation of sows comprises the following steps:

1) Weighing qualified corns, bean pulp, rice bran meal, DDGS, corn germ meal, corn husks, stone powder and senna leaves according to the parts by weight, crushing to obtain raw materials with the particle size of 2.0mm, putting the raw materials into a stirring tank, stirring at the stirring speed of 30-50r/m, and mixing for 10-20 minutes to obtain a dry material with the variation coefficient of less than 7%.

2) Preparing an aerobic fermentation agent:

Preparation of bacillus subtilis: inoculating activated bacillus subtilis into a 100mL conical flask filled with 50mL of common broth culture medium in a sterile laboratory, and culturing for 8-14 h at 35-38 ℃ to obtain liquid bacillus bacterial liquid, wherein the number of viable bacteria in the bacterial liquid is more than or equal to 10 ⁹ cfu/mL;

preparation of agrocybe cylindracea fungi: under the aseptic condition, transferring activated agrocybe cylindracea strain to PAD slant culture medium (pH 5.5-6.5), and culturing in an incubator at 20-27 ℃ under the strain culturing condition: intermittently ventilating the mycelium growing chamber for 1-2 times per day, wherein the relative humidity is preferably 65% -70%; the mycelium grows up the inclined plane to be the agrocybe cylindracea fungus strain.

Preparation of straw mushroom fungi: under aseptic condition, inoculating activated straw mushroom strain to glass test tube PAD culture medium, culturing at 10-40 deg.C for 3-5 days until mycelium grows up to 15-20 deg.C.

The bacillus subtilis, agrocybe cylindracea fungi and straw mushroom fungi are mixed according to the weight ratio of 1:1: 1.

3) Diluting the aerobic fermentation agent 100-250 times into clean water, stirring uniformly, pumping the aerobic fermentation agent into the raw material stirring tank of the step 1), mixing for 5-10 minutes to uniformly distribute bacterial liquid and raw material components, preparing a wet material I, putting the wet material I into a feed bag, warehousing and standing for ventilation fermentation, wherein the warehouse environment temperature is 23-28 ℃, and the fermentation time is as follows: fermenting for 3-5 days in winter and fermenting for 2-3 days in other seasons.

4) The preparation method of the anaerobic fermentation agent comprises the following steps:

Preparation of enterococcus faecalis: under the aseptic condition, the activated enterococcus faecalis is inoculated in an MRS culture medium, and is cultured for 13 to 16 hours at the temperature of between 35 and 37 ℃ in an anaerobic incubator, and the number of viable bacteria in bacterial liquid is more than or equal to 10 ¹⁰ cfu/mL.

Preparation of Saccharomyces cerevisiae: under the aseptic condition, inoculating the activated saccharomyces cerevisiae into PAD liquid culture medium, culturing for 10-16 h at 28-32 ℃ in an anaerobic incubator, wherein the number of viable bacteria in the bacterial liquid is more than or equal to 10 ¹⁰ cfu/mL.

The weight ratio of enterococcus faecalis to saccharomyces cerevisiae is 2:1.

5) Diluting the anaerobic fermentation agent into clean water by 30-100 times, stirring uniformly, putting the fermented wet material I into a stirring tank, stirring at a stirring speed of 30-40 r/m, mixing for 10-20 minutes, pumping the anaerobic fermentation agent into the fermented wet material I, mixing the anaerobic fermentation agent bacterial liquid and the fermented wet material I uniformly for 5-10 minutes again to obtain a wet material II, putting the wet material II into a breathing bag with an exhaust hole, sealing a bag opening, warehousing, standing and fermenting, stacking layers of the wet material I is less than or equal to 5, and the ambient temperature of a storehouse is more than or equal to 25 ℃, wherein the fermentation time is as follows: fermenting for 2-3 days in winter and fermenting for 1-2 days in other seasons.

Example 2:

microbial fermented feed for preventing and treating constipation of sow and preparation method thereof

(1) Preparing dry materials: the raw material components in weight ratio are as follows: is prepared from 30 parts of corn, 25 parts of bean pulp, 5 parts of DDGS, 10 parts of corn gluten feed, 9.5 parts of corn germ meal, 18 parts of rice bran meal, 2 parts of stone powder and 0.5 part of senna leaf. The dry mixed feed is crushed by sieve holes of a hammer mill, and the sieve holes of the hammer mill have the diameter of 2.0mm.

(2) 1.5Kg of aerobic fermentation agent (Bacillus subtilis: agrocybe cylindracea: straw mushroom fungi=1:1:1) was added to 200kg of clean water and stirred to uniformly disperse the bacterial liquid components, and the preparation method was as described in example 1.

(3) 800Kg of the dry material mixture was weighed and put into a stirring tank to be stirred at a stirring speed of 40r/m for 20 minutes. And (3) pumping the aerobic fermentation agent into the dry material and mixing for 8 minutes again to ensure that all components of the bacterial liquid and the dry material can be uniformly distributed.

(4) And filling the mixed wet materials into a feed bag. The feed bags are put in storage for standing fermentation, and ventilation fermentation is carried out for 3 days.

(5) 1.5Kg of anaerobic fermentation agent (enterococcus faecalis: saccharomyces cerevisiae=2:1) was added to 50 pure water and stirred as in example 1, and was pumped into the fermented wet material obtained in step (4), stirred in a stirring tank at a stirring speed of 30r/m, and mixed for 20 minutes.

(6) And (3) filling the mixed wet materials into a breathing bag with an exhaust hole, and sealing the opening of the bag. Placing in a warehouse for standing and fermenting for 2 days.

The use method of the fermented feed comprises the following steps: the microbial fermentation feed is uniformly mixed with the conventional feed, and the proportion of the microbial fermentation feed is 1-3: 20, can be fed at each stage of sow feeding. The constipation prevention is fed after being evenly mixed with the conventional batch according to the proportion of 5 to 10 percent; the constipation of the sow is treated according to the proportion of 10 to 15 percent and is even with the conventional feed. Continuous feeding for 3-5 days, such as relieving, and then reducing the dosage to within 10%. And the sow can also be fed by adopting an additional feeding mode, and each sow is fed with 100-250 g of fermented feed per day.

Example 3:

method for preparing microbial fermented feed for preventing and treating constipation of sow

(1) Preparing dry materials: the raw material components in weight ratio are as follows: the corn gluten feed comprises the following components in percentage by mass of 28 percent of corn, 26 percent of bean pulp, 5 percent of DDGS, 10 percent of corn gluten feed, 11 percent of corn germ meal, 17 percent of rice bran meal, 2 percent of stone powder and 1 percent of senna leaf. The dry mixed feed is crushed by sieve holes of a hammer mill, and the sieve holes of the hammer mill have the diameter of 2.0mm.

(2) 1.5Kg of aerobic fermentation agent (bacillus subtilis: agrocybe cylindracea: straw mushroom fungi=1:1:1, preparation method see example 1) was added to 250kg of clean water and stirred to uniformly disperse the bacterial liquid components.

(3) The 850 dry mix was weighed and placed into a stirred tank for stirring. The stirring speed is 40r/m, and the mixture is mixed for 20 minutes. And (3) pumping the aerobic fermentation agent into the dry material and mixing for 10 minutes again to ensure that all components of the bacterial liquid and the dry material can be uniformly distributed.

(4) And filling the mixed wet materials into a feed bag. And (5) warehousing, standing and fermenting the feed bags. Fermenting for 2 days by ventilation.

(5) 2Kg of anaerobic fermentation agent (enterococcus faecalis: saccharomyces cerevisiae=2:1, preparation method thereof is described in example 1) was added to 80kg of clean water and stirred, which was pumped into the above fermented feed, stirred in a stirring tank at a stirring speed of 30r/m, and mixed for 15 minutes.

The use method of the fermented feed comprises the following steps: as in example 2.

Example 4:

Microbial fermentation feed method for preventing and treating constipation of sow

(1) Preparing dry materials: the raw material components in weight ratio are as follows: the components and mass percentages of the corn and soybean meal are 30 percent, 25 percent, DDGS 4 percent, 10 percent of corn gluten feed, 9 percent of corn germ meal, 19.5 percent of rice bran meal, 2 percent of stone powder and 0.5 percent of senna leaf. Pulverizing the dry mixed feed with hammer mill sieve mesh, and mixing. The diameter of the sieve pore of the hammer mill is 2.0mm.

(2) 1.5Kg of aerobic fermentation agent (bacillus subtilis: agrocybe cylindracea: straw mushroom fungi=1:1:1, preparation method see example 1) was added to 200kg of clean water and stirred to uniformly disperse the bacterial liquid components.

(3) And weighing 750 the dry material mixture, and putting the dry material mixture into a stirring tank for stirring. The stirring speed is 40r/m, and the mixture is mixed for 20 minutes. And (3) pumping the aerobic fermentation agent into the dry material and mixing for 5 minutes again to ensure that all components of the bacterial liquid and the dry material can be uniformly distributed.

(4) And filling the mixed wet materials into a feed bag. And (5) warehousing, standing and fermenting the feed bags. Fermenting for 3 days by ventilation.

(5) 1.5Kg of anaerobic fermentation agent (enterococcus faecalis: saccharomyces cerevisiae=2:1, preparation method thereof is described in example 1) was added to 80 pure water and stirred, which was pumped into the above fermented feed, stirred in a stirring tank at a stirring speed of 30r/m, and mixed for 15 minutes.

Example 5

The invention is further described in connection with feeding trials and clinical applications as follows:

the use effect of the microbial fermented feed of the present invention was evaluated by in vitro fermented feed quality detection and sow constipation analysis over 6 months.

Quality detection of microbial fermented feed

1.1 Test materials and methods

1.1.1 Selection of test feeds

The microbial fermented feed preparation process of example 1 is adopted, and 10kg of fermented feed is randomly taken from different parts in a feed bag after 4-5 days of fermentation.

1.1.2 Measurement index

1.1.2.1 Sensory evaluation the appearance and colour of the microbial fermented feed were observed and the mould of the feed was checked. The fermentation condition of the feed is analyzed by measuring the smell of the feed. And touching the feed to observe the hardness of the feed.

1.1.2.2 Determination of moisture in microbial fermented feed

The original sample is contracted to 500g by a quartering method, is crushed to 40 meshes after being air-dried, is contracted to 200g by the quartering method, is filled into a sealed container, and is stored in a shade and dry place. The fermented feed is dried in a drying oven at 105 ℃ for 3 hours, cooled in a dryer for 30 minutes, weighed, dried in the drying oven at 105 ℃ for 1 hour, and weighed to constant weight. For each sample, two replicates were taken and measured as arithmetic mean. The measured values of the two replicates differ by no more than 0.002g, which is a constant weight. The calculation formula is as follows:

moisture (%) = [ (W1-W2)/(W1-W0) ] x 100 ]

Wherein: w1-105 ℃ before drying the sample and weighing the sample dish, g;

w2-drying at-105 ℃ and weighing the sample and the sample dish, and g;

w0-weight-the weight of the dish was constant, g.

Determination of the pH of 1.1.2.3 microbial fermented feed

The fermented feed sample is mixed according to the proportion, passes through a 40-mesh sieve, 100.0g of the fermented feed sample is accurately weighed by an electronic balance and placed in a 500mL beaker, 200mL of deionized water is added, the solution is uniformly stirred by a magnetic stirrer, the pH of the solution is measured by an acidometer, and the data is recorded as the feed pH after the pH is constant.

1.1.2.4 Determination of feed acid binding force

The microbial fermented feed of example 1 was added to a conventional feed at a ratio of 0% (control), 5%, 10%, 15% and mixed uniformly. Sampling by a quartering method, crushing, sieving with a 60-mesh sieve, accurately weighing 100.0g by an electronic balance, placing in a 500mL beaker, adding 200mL deionized water, uniformly stirring by a magnetic stirrer, fully soaking the sample (room temperature for 20-30 minutes), titrating by 1moL/L hydrochloric acid, fully stirring, standing for 10 minutes, measuring the pH value of the solution by an acidometer, and recording the volume of the hydrochloric acid solution with consumption standard of 1.00moL/L when the pH value is constant to 4, thus obtaining the feed acid binding force.

1.2 Results and analysis

1.2.1 Sensory evaluation results

The color of the microbial fermentation feed is relatively close to the original color, the microbial fermentation feed has rich ferment flavor, and the micro-grabbing feed has loose, soft and slightly moist feeling, so that the microbial fermentation feed has no mildew, proper smell and moderate moisture. Meets the fermentation requirement of microbial fermentation feed.

1.2.2 Moisture content results of microbial fermented feed

The moisture content of the microbial fermented feed of the invention was 31%.

1.2.3 PH value of fermented feed

The pH of the microbial fermented feed of the present invention was 4.4, indicating that the acid bacteria of the microbial fermented feed of the present invention became dominant bacteria. The acidity is optimal for sow intake, and has important significance for digestion and absorption of sows and improvement of intestinal microecological environment.

1.2.4 Determination of feed acid binding force

TABLE 1 acid binding force of microbial fermented feed (mmoL HCL/100 g)

	0.0% (Control)	5％	10％	15％
					Acid binding force	34.0±0.26	31.8±0.32	30.0±0.18	26.2±0.26

With the increase of the microbial fermentation feed additive amount, the smaller the feed acid binding force is. The acid binding force of the feed is an index of the degree to which the feed adsorbs free acid in the stomach. The acid binding force of the feed mainly depends on the proportion and the content of the feed components. The microbial fermentation feed is acidic, and the microbial fermentation feed is added into the feed, so that the pH of the feed can be effectively reduced, the acid binding force of the feed is reduced, the influence on the acidity of the digestive tract of pigs is reduced, a proper digestion internal environment is provided, and the optimal pH of pepsin is maintained.

(II) test of microbial fermentation feed for improving constipation and feed intake of sow

2.1 Materials and methods

2.1.1 Feed and animals

20 Constipation sows are randomly selected and fed with a microbial fermented feed containing 15% (namely, 15% of the fermented feed of the embodiment 2 is added into basic ration).

In addition, 30 healthy sows with the same gestation time and about 110 days of gestation with the expected period (within seven days) are randomly selected and divided into two groups, wherein the first group is a control group, basic daily ration is fed, the second group is a fermented feed group, and 8% of microbial fermented feed is added into the basic daily ration (microbial fermented feed shown in the example 3 is selected).

2.1.2 Index determination

2.1.2.1 Constipation of sow

The property of the sow feces is observed once a day in the morning and evening. Constipation of sow is recorded according to the grade of fecal character (0: no fecal discharge; I: constipation (granule; II: firm; III: light firm; IV: soft; V: diarrhea).

2.1.2.2 Determination of sow production Performance

Recording the feed intake of 30 sows in the late childbirth and lactation period, and calculating the average daily feed intake.

And (5) measuring the survival rate of the piglets at birth, the average daily gain of the weaned piglets and the weaned survival rate.

2.2 Results and analysis

2.2.1 Effect of microbial fermented feed on constipation in sow

At the beginning of the test, constipation sows were rated at grade i and grade ii (fig. 1), and the feces were secret and firm. By feeding the microbial fermented feed, the properties of the feces are improved over 3-5 days (figure 2). The feces gradually soften and become soft. Constipation of 20 sows is completely relieved or cured, and diarrhea does not occur. The microbial fermentation feed disclosed by the invention improves the internal micro-ecological environment of intestinal tracts of constipation sows, and prevents constipation.

2.2.2 Influence of microbial fermented feed on sow feed intake

As can be seen from the data in Table 2, the feed intake of sows fed the microbial fermented feed was significantly increased (P < 0.05) both in the late gestation period and in the breast-feeding period compared to the control group. The acidic product produced by fermenting the microbial fermented feed has mellow fragrance, improves the palatability of the feed, not only improves the feed intake of sows, but also is beneficial to improving the intestinal microecological environment of the sows.

Table 2 sow performance measurements

Index (I)	Control group	Fermented feed group
			Feed intake (kg) in the late gestation period	3.26±0.28^b	3.89±0.31^a
Daily feed intake (kg) in lactation period	5.91±0.47^b	6.24±0.48^a
			Survival rate of piglets at birth (%)	95.28±2.45^b	100±0.00^a
Average daily gain (kg) of weaned pigs	8.96±0.56^b	9.89±0.78^a
			Weaning survival rate (%)	90.25±0.78^b	98.26±0.85^a

Note that: the same row of values shoulder-annotated with different letters indicates significant differences (P < 0.05), the same letters indicate insignificant differences (P > 0.05).

The test result shows that the survival rate of the piglets in the fermented feed group is obviously higher than that in the control group (P is less than 0.05). The microbial fermentation feed provided by the invention contains mellow taste after microbial fermentation, so that appetite of sows can be promoted, meanwhile, after the feed is fermented, components in the feed are changed, and a large amount of metabolites are produced by the raw materials of the feed and after fermentation, so that nutritional values of vitamins and amino acids of the raw materials and the microorganisms are ensured, and nutritional supply and support of reproductive performance of the sows are ensured. In addition, the survival rate of weaned pigs and the average daily gain of weaned pigs of the fermented feed group are also obviously higher than those of a control group (P is less than 0.05), and are respectively 10.4 percent and 8.9 percent higher than those of the control group. The microbial fermentation feed disclosed by the invention can increase the survival of beneficial bacteria in pigs, adjust the balance of micro-ecology in the intestinal tracts and the environment of sows, has advantages of beneficial bacteria, and reduces the chances of piglets contacting with pathogenic bacteria such as escherichia coli, so that the morbidity and death of the piglets are reduced, and the survival rate of the piglets is improved. Meanwhile, the fermented feed contains a large amount of biological probiotics metabolites (amino acids, vitamins, active skin and other probiotics), after the sow eats the fermented feed, the non-specific resistance of the piglet is improved by milk, and the disease resistance and the nutrition absorption rate of the piglet are obviously improved, so that the survival rate and the weight of weaned piglets in the fermented feed group are higher than those of weaned piglets in the control group.

According to the nutrition physiological characteristics of the sows, the microbial fermentation feed disclosed by the invention is used for optimizing the special requirements of the sows on daily ration protein, energy and crude fiber by using various feed raw materials according to the nutrition physiological characteristics of the sows, so that the special requirements of the sows on daily ration protein, energy and crude fiber are met, the intestinal peristalsis of the sows is promoted, the digestion function of the dry feed is improved, the occurrence of constipation is fundamentally prevented, the cure rate reaches 95%, and the constipation is basically eliminated after continuous feeding for 3-5 days.

The foregoing detailed description of the preparation and application of the present invention is merely for illustrating the technical scheme of the present invention, and is not intended to limit the scope of the patent claims; modifications of the technical solutions described in the foregoing embodiments, or equivalent substitutions of some of the technical features thereof, or direct or indirect application in other relevant technical fields are included in the scope of the technical solutions claimed in the present invention.

Claims

1. The microbial fermented feed for preventing constipation of the sow is characterized by being prepared from the following raw materials in parts by weight: 28-31 parts of corn, 25-27 parts of bean pulp, 4-6 parts of DDGS, 9-12 parts of corn gluten feed, 9-11 parts of corn germ meal, 17-19 parts of rice bran meal, 1.5-2.5 parts of stone powder, 0.3-1 part of senna leaf, 0.1-0.3 part of aerobic fermentation agent and 0.1-0.3 part of anaerobic fermentation agent;

the aerobic fermentation agent consists of bacillus subtilis, agrocybe cylindracea fungi and straw mushroom fungi; the weight ratio of the bacillus subtilis to the agrocybe cylindracea to the straw mushroom is 1:1:1, a step of; the preservation number of the agrocybe cylindracea fungi is CGMCC No.16085; the preservation number of the straw mushroom fungi is CGMCC No.16086;

the anaerobic fermentation agent consists of enterococcus faecalis and saccharomyces cerevisiae; the dosage ratio of enterococcus faecalis to Saccharomyces cerevisiae is 2:1;

the preparation method of the microbial fermentation feed comprises the following steps:

1) Crushing the corn, the bean pulp, the rice bran meal, the DDGS, the corn germ meal, the corn gluten feed, the stone powder and the senna to obtain raw materials with the particle size of 2.0mm, putting the raw materials into a stirring tank for stirring and mixing to obtain dry materials, wherein the variation coefficient is less than 7%; the stirring rotating speed is 30-50 r/m; the mixing time is 10-20 minutes;

2) Preparation of aerobic fermentation agent

Preparation of bacillus subtilis: inoculating activated bacillus subtilis into a 100mL conical flask filled with 50mL of common broth culture medium in a sterile laboratory for culturing to obtain liquid bacillus bacterial liquid, wherein the number of viable bacteria in the bacterial liquid is more than or equal to 10 ⁹ cfu/mL; the temperature of the culture is 35-38 ℃ and the time is 8-14 h;

Preparation of agrocybe cylindracea fungi: under aseptic condition, transferring activated agrocybe cylindracea strain onto PAD slant culture medium, and culturing in incubator under strain culture condition: intermittently ventilating the mycelium growing chamber for 1-2 times per day, wherein the relative humidity is preferably 65% -70%; the mycelium grows up the inclined plane to be the agrocybe cylindracea fungus strain; the pH value of the PAD slant culture medium is 5.5-6.5; the temperature of the culture is 20-27 ℃;

preparation of straw mushroom fungi: inoculating activated straw mushroom strains to a glass test tube PAD culture medium for culturing under a sterile condition, and storing after 3-5 days when mycelia grow on the culture medium; the temperature of the culture is 10-40 ℃; the storage temperature is 15-20 ℃;

3) Diluting the aerobic fermentation agent into clean water, uniformly stirring, pumping the aerobic fermentation agent into the raw material stirring tank in the step 1), uniformly distributing bacterial liquid and raw material components, preparing a wet material I, putting the wet material I into a feed bag, warehousing and standing for ventilation fermentation, wherein the temperature of a warehouse environment is 23-28 ℃; the fermentation time: fermenting for 3-5 days in winter and fermenting for 2-3 days in other seasons; the dilution multiple is 100-250 times; the mixing time is 5-10 minutes;

4) Preparation of anaerobic fermentation agent

Preparation of enterococcus faecalis: inoculating the activated enterococcus faecalis into MRS culture medium under aseptic condition, culturing in anaerobic incubator, and culturing in bacterial liquid with viable count of not less than 10 ¹⁰ cfu/mL; the temperature of the culture is 35-37 ℃ and the time is 13-16 h;

Preparation of Saccharomyces cerevisiae: inoculating activated saccharomyces cerevisiae into PAD liquid culture medium under aseptic condition, culturing in anaerobic incubator, wherein the number of viable bacteria in bacterial liquid is more than or equal to 10 ¹⁰ cfu/mL; the temperature of the culture is 28-32 ℃ and the time is 10-16 h;

The enterococcus faecalis and the saccharomyces cerevisiae are mixed according to the weight ratio of 2:1, mixing;

5) Diluting the anaerobic fermentation agent into clean water, stirring uniformly, putting the fermented wet material I into a stirring tank, stirring, mixing, pumping the anaerobic fermentation agent into the fermented wet material I, mixing again to uniformly mix the anaerobic fermentation agent bacterial liquid and the fermented wet material I to obtain a wet material II, putting the wet material II into a breathing bag with an exhaust hole, sealing a bag opening, warehousing, standing and fermenting, stacking the layers to be less than or equal to 5, and keeping the ambient temperature of a warehouse to be more than or equal to 25 ℃, wherein the fermentation time is as follows: fermenting for 2-3 days in winter and fermenting for 1-2 days in other seasons; the dilution multiple is 30-100 times; the stirring speed in the stirring tank is 30-40 r/m, and the mixing time is 10-20 minutes; the time for the remixing is 5-10 minutes.

2. The microbial fermented feed according to claim 1, wherein the microbial fermented feed for preventing constipation of sows is prepared from the following raw materials in parts by weight: 30 parts of corn, 25 parts of bean pulp, 5 parts of DDGS, 10 parts of corn gluten feed, 9.4 parts of corn germ meal, 18 parts of rice bran meal, 2 parts of stone powder, 0.6 part of senna leaf, 0.2 part of aerobic fermentation agent and 0.2 part of anaerobic fermentation agent.

3. The microbial fermented feed according to claim 1, wherein the microbial fermented feed quality specifications: the pH value of the fermented feed is 5.5-6; the moisture of the fermented feed is 30% +/-2%; the hygienic index of the fermented feed meets the requirements of the hygienic standard of the feed; the raw materials and the production links of the microbial fermentation feed cannot contain any antibiotics.

4. A process for the preparation of a microbial fermented feed according to any one of claims 1 to 3, comprising the steps of:

2) Preparation of aerobic fermentation agent

4) Preparation of anaerobic fermentation agent