CN112471353A - Long-acting symbiotic composite liquid microecological preparation for poultry feeding and preparation method thereof - Google Patents
Long-acting symbiotic composite liquid microecological preparation for poultry feeding and preparation method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
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Abstract
The invention belongs to the technical field of microecologics, and provides a long-acting symbiotic composite liquid microecologics for poultry feeding, which comprises 5-10 parts of lactobacillus plantarum LP45, 1-5 parts of bacillus subtilis, 1-5 parts of bacillus licheniformis, 2-3 parts of saccharomyces cerevisiae, 11-16 parts of brown granulated sugar and 30-40 parts of water. By the technical scheme, the problem that the survival rate of viable bacteria of the microecological preparation in an animal organism is poor in the prior art, so that the effect of adjusting the intestinal tract is poor is solved.
Description
Technical Field
The invention belongs to the technical field of microecologics, and relates to a long-acting symbiotic composite liquid microecologics for poultry feed and a preparation method thereof.
Background
In recent years, the livestock industry in China has been greatly developed, but with the development of the livestock breeding industry and the improvement of the intensification degree of the livestock breeding industry, various problems are caused by the overuse of antibiotics, the breeding environment is worsened, the variation rate of pathogens is improved, symptoms are atypical, old diseases are new, new diseases are frequent, and the control is more and more difficult.
Common diseases of poultry comprise escherichia coli, enteritis, salpingitis and the like, but the current common solution is to use antibiotics, the phenomenon of unreasonable use of antibiotics also occurs frequently, the antibiotics play a role and play a role in screening drug-resistant bacteria, according to the data detected by a bacterial drug-resistant detection network in China, staphylococcus has over 50 percent of drug resistance to various common antibiotics such as methicillin, macrolides, quinolones and the like in over 100 domestic bacteria, and the drug resistance of escherichia coli to antibiotic drugs is over 50 percent.
At present, the nation has strict rules on the use of antibiotics, and aims to prevent feed from resisting, limit cultivation and prevent food from resisting. In 2018, the work scheme of animal antibacterial use reduction action trial point issued by rural agricultural departments (2018-2021) defines the time schedule of feed-end resistance reduction and resistance limitation, and the feed resistance is imminent.
Importance of the intestinal microecological balance of poultry: the intestinal tract is not only the main site for the digestion and absorption of nutrients in the body, but also the congenital barrier for the body to resist external pathogenic microorganisms and their harmful metabolites. The intestinal barrier consists of a microbial barrier, a chemical barrier, a physical barrier and an immune barrier in sequence from the basal side of the intestinal lumen, and different barriers have different biological functions and molecular regulation mechanisms.
The intestinal tract of healthy animals has a large amount of bacteria to form a microecosystem of the intestinal tract of the animals, the normal intestinal tract function is the guarantee of the health of animal organisms and is the key for improving the growth performance of poultry, and the intestinal barrier of the poultry is damaged, so that the health of the animals is influenced by different degrees. The probiotics can inhibit the growth and adhesion of pathogenic bacteria in the intestinal tract by means of colonizing and resisting, dominating population action, generating bacteriostatic metabolites, promoting body immunity and the like, and establish the dominant position of the probiotics, thereby maintaining the microecological balance in the intestinal tract. The survival rate of viable count in animal bodies of the existing microecological preparation is poor, so that the effect of regulating intestinal tracts is poor.
Disclosure of Invention
The invention provides a long-acting symbiotic composite liquid microecological preparation for feeding poultry and a preparation method thereof, and solves the problem that the survival rate of viable bacteria of the microecological preparation in an animal body is poor in the prior art, so that the effect of regulating intestinal tracts is poor.
The technical scheme of the invention is realized as follows: the long-acting symbiotic composite liquid microecological preparation for poultry feed comprises, by mass, 5-10 parts of lactobacillus plantarum LP45, 1-5 parts of bacillus subtilis, 1-5 parts of bacillus licheniformis, 2-3 parts of saccharomyces cerevisiae, 11-16 parts of brown granulated sugar and 30-40 parts of water.
Further, the viable count of the long-acting symbiotic composite microecological preparation is more than or equal to 5 x 108cfu/mL。
The preparation method of the long-acting symbiotic composite liquid microecological preparation for poultry feeding comprises the following steps:
A. preparing each component according to the formula of the long-acting symbiotic composite microecological preparation for feeding the poultry;
B. inoculating lactobacillus plantarum LP45 into a fermentation medium a, mixing, performing anaerobic fermentation at 37 ℃ for 16 hours, and controlling the pH value to be 5.0 to obtain lactobacillus plantarum LP45 fermentation liquor;
respectively inoculating bacillus subtilis, bacillus licheniformis and saccharomyces cerevisiae into the fermentation culture medium b for mixing, and carrying out aerobic fermentation at 37 ℃ for 48h to obtain micro-fermentations of the bacillus subtilis, the bacillus licheniformis and the saccharomyces cerevisiae;
C. and D, adding brown granulated sugar into the lactobacillus plantarum LP45 fermentation liquor obtained in the step B, adding micro-fermentation products of bacillus subtilis, bacillus licheniformis and saccharomyces cerevisiae, mixing, performing anaerobic fermentation at 37 ℃ for 85 hours, stirring for 5min every 5 hours, reducing the pH value to 4.0, and stopping fermentation.
Further, the inoculation amount of the lactobacillus plantarum LP45 in the step B is 5% of the mass of the fermentation medium a.
And further, the inoculation amount of the bacillus subtilis and the bacillus licheniformis in the step B is 5% of the mass of the spore culture medium, and the inoculation amount of the saccharomyces cerevisiae is 5% of the mass of the fermentation culture medium B.
Further, the fermentation medium a in the step B comprises 1% of beef extract powder, 0.5% of yeast extract, 1% of peptone, 2% of glucose and K in percentage by mass2HPO40.2%, sodium acetate 0.5%, MgSO4·7H2O 0.02％，MnSO4·4H20.005% of O, 80.1% of Tween, 0.2% of triammonium citrate and the balance of water; the fermentation medium a is sterilized, and the pH value is 6.2-6.6 before sterilization.
Further, the fermentation medium B in the step B comprises 1% of yeast extract powder, 2% of peptone, 2% of glucose and the balance of water in percentage by mass; sterilizing the fermentation culture medium b, wherein the pH value is 6.2-6.6 before sterilization;
according to the mass percentage, the spore culture medium in the step B comprises 1% of peptone, 0.3% of beef extract powder, 0.5% of sodium chloride and the balance of water; the spore culture medium is sterilized, and the pH value is 7.0-7.3 before sterilization.
Further, when the pressure in step C is higher than 0.1 MPa, the exhaust pressure is reduced to 0.025 MPa.
The working principle and the beneficial effects of the invention are as follows:
1. after the microecological preparation product is treated under the acidic condition for 4 hours, the survival rate of the microecological preparation product is more than 90 percent, and the microecological preparation product can play a role in colonization and probiotics in intestinal tracts; the survival rate is 80 percent after the treatment of 0.3 percent of the ox bile salt, enough viable bacteria still reach the intestinal tract for permanent planting, the probiotic effect is exerted, the physique of the broiler chicken can be obviously improved, the health condition of the broiler chicken is improved, the morbidity of the broiler chicken is reduced, and the problem of poor intestinal tract regulation effect is solved.
2. According to the invention, through a specific preparation process of the long-acting symbiotic composite liquid microecological preparation, the synergistic effect of the strains is better exerted, the protease and the amylase generated by the lactobacillus plantarum with high enzyme activity are used for better improving the conversion rate of poultry feed, reducing the feed conversion ratio, improving the immunity and disease resistance of the organism, and enabling chickens to show red crowns and have strong activity.
3. The strain collection base of the invention comprises: the strain is derived from more than 30 areas of healthy animals and excrement such as Sichuan, Yunnan, Tibet, Xinjiang, inner Mongolia and the like; according to comprehensive analysis of strain cell morphology, physiological and biochemical characteristics, a 16S rRNA gene sequence, a pheS gene sequence and the like, LP45(YMC1005) is identified as Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus plantarum LP45(YMC1005) with the patent preservation number of CGMCC No.8072, has a probiotic flora with intestinal mucosa colonization capacity, has strong adhesion, plays a role in competitive exclusion of pathogenic microorganisms, establishes and maintains a normal dominant flora of the intestinal tract, and adjusts intestinal tract balance. The lactobacillus plantarum can synthesize various vitamins and nutrient elements, generate various digestive enzymes, improve the utilization rate of the feed, generate organic acid and promote the absorption of nutrient substances; it is also effective in preventing gastrointestinal inflammation, gastrointestinal allergy, pathogenic bacteria infection, endogenous infection and antibiotic-associated diarrhea.
The action mechanism of the long-acting symbiotic composite liquid microecological preparation product is as follows:
A. theory of dominant flora: acid metabolites such as lactic acid, formic acid, acetic acid, propionic acid, and butyric acid are produced. The acidic substance of the acid substance reduces the pH value of the intestinal tract, effectively inhibits the growth and reproduction of various pathogenic bacteria, has no influence on surrounding probiotics, and finally establishes a micro-ecological environment taking the probiotics as a leading position in the intestinal tract.
B. The theory of biological antagonism: the hydrogen peroxide is generated in the metabolic process, a peroxidase-thiocyanate reaction system in the intestinal tract can be activated, the lactic acid peroxidase is combined with the hydrogen peroxide, and then the thiocyanate is oxidized into an oxidative intermediate product to inhibit the growth and reproduction of pathogenic bacteria.
C. The theory of toxin clearance: metabolism can produce bacteriocins such as pediocin, streptococcin and the like, and can enter the interior of the thallus of a pathogen to destroy genetic materials or important metabolic pathways of the pathogen.
Drawings
The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.
FIG. 1 shows the bacteriostatic effect of SD-3301 products of example 1 of the present invention on Escherichia coli.
FIG. 2 shows the bacteriostatic effect of the SD-3301 product of the invention on Salmonella pullorum.
FIG. 3 shows the shigella dysenteriae-inhibiting effect of SD-3301 products of the present invention.
FIG. 4 shows the bacteriostatic effect of the SD-3301 product of the invention on Salmonella enteritidis.
FIG. 5 shows the survival of the SD-3301 product of example 1 under ph 3.0.
FIG. 6 shows the survival of the product SD-3301 of example 1 in the presence of 0.3% bovine bile salt.
FIG. 7 shows the stability of viable count of SD-3301 products of the present invention in example 1.
FIG. 8 shows the influence of intestinal structures of broilers in experimental group and control group; wherein (1) represents the culture of a control group 21d, (2) represents the culture of an experimental group 21d, (3) represents the culture of a control group 42d, and (4) represents the culture of an experimental group 42 d.
FIG. 9 shows excrements of broilers before use in the experimental group of the present invention.
FIG. 10 shows excrements of broiler chickens after use in experimental groups of the present invention.
FIG. 11 shows the effect of the experimental group and the control group of the present invention on the weight gain of broiler chickens.
FIG. 12 shows the effect of the experimental group and the control group on the feed-meat ratio of broiler chickens.
FIG. 13 shows the effect of the experimental group and the control group on the survival rate of broiler chickens.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. The media used in the following examples were prepared as follows: according to the mass percentage, the fermentation medium a comprises 1 percent of beef extract powder, 0.5 percent of yeast extract, 1 percent of peptone, 2 percent of glucose and K2HPO40.2%, sodium acetate 0.5%, MgSO4·7H2O 0.02％，MnSO4·4H20.005% of O, 80.1% of Tween, 0.2% of triammonium citrate and the balance of water; sterilizing the fermentation culture medium a, wherein the pH value is 6.2-6.6 before sterilization; the fermentation medium b comprises 1% of yeast extract powder, 2% of peptone, 2% of glucose and the balance of water; sterilizing the fermentation culture medium b, wherein the pH value is 6.2-6.6 before sterilization; the spore culture medium comprises peptone 1%, beef extract powder 0.3%, sodium chloride 0.5% and water in balance; the spore culture medium is sterilized, and the pH value is 7.0-7.3 before sterilization.
Detailed description of the preferred embodiments
Example 1
The preparation method of the long-acting symbiotic composite liquid microecological preparation for poultry feeding comprises the following steps:
A. preparing each component according to the formula of the long-acting symbiotic composite liquid microecological preparation for feeding the poultry in the table 1;
B. inoculating lactobacillus plantarum LP45 into a fermentation medium a, mixing, performing anaerobic fermentation at 37 ℃ for 16 hours, wherein the inoculation amount is 5% of the mass of the fermentation medium a, and the pH value is controlled at 5.0, so as to obtain lactobacillus plantarum LP45 fermentation liquor;
inoculating bacillus subtilis and bacillus licheniformis to a spore culture medium, inoculating saccharomyces cerevisiae to a fermentation culture medium b, and performing aerobic fermentation at 37 ℃ for 48h to obtain micro-fermentations of the bacillus subtilis, the bacillus licheniformis and the saccharomyces cerevisiae; wherein the inoculation amount of the bacillus subtilis and the bacillus licheniformis is 5% of the mass of the spore culture medium, and the inoculation amount of the saccharomyces cerevisiae is 5% of the mass of the fermentation culture medium b;
C. and D, adding brown granulated sugar into the lactobacillus plantarum LP45 fermentation liquor obtained in the step B, adding micro-fermentation products of bacillus subtilis, bacillus licheniformis and saccharomyces cerevisiae, mixing, performing anaerobic fermentation at 37 ℃ for 85 hours, stirring for 5min every 5 hours, and stopping fermentation when the pressure is higher than 0.1 MPa, the exhaust pressure is reduced to 0.025 MPa, and the pH value is reduced to 4.0.
Example 2
The preparation method of the long-acting symbiotic composite liquid microecological preparation for poultry feeding comprises the following steps:
A. preparing each component according to the formula of the long-acting symbiotic composite liquid microecological preparation for feeding the poultry in the table 1;
B. inoculating lactobacillus plantarum LP45 into a fermentation medium a, mixing, performing anaerobic fermentation at 37 ℃ for 16 hours, wherein the inoculation amount is 5% of the mass of the fermentation medium a, and the pH value is controlled at 5.0, so as to obtain lactobacillus plantarum LP45 fermentation liquor;
inoculating bacillus subtilis and bacillus licheniformis to a spore culture medium, inoculating saccharomyces cerevisiae to a fermentation culture medium b, and performing aerobic fermentation at 37 ℃ for 48h to obtain micro-fermentations of the bacillus subtilis, the bacillus licheniformis and the saccharomyces cerevisiae; wherein the inoculation amount of the bacillus subtilis and the bacillus licheniformis is 5% of the mass of the spore culture medium, and the inoculation amount of the saccharomyces cerevisiae is 5% of the mass of the fermentation culture medium b;
C. and D, adding brown granulated sugar into the lactobacillus plantarum LP45 fermentation liquor obtained in the step B, adding micro-fermentation products of bacillus subtilis, bacillus licheniformis and saccharomyces cerevisiae, mixing, performing anaerobic fermentation at 37 ℃ for 85 hours, stirring for 5min every 5 hours, and stopping fermentation when the pressure is higher than 0.1 MPa, the exhaust pressure is reduced to 0.025 MPa, and the pH value is reduced to 4.0.
Example 3
The preparation method of the long-acting symbiotic composite liquid microecological preparation for poultry feeding comprises the following steps:
A. preparing each component according to the formula of the long-acting symbiotic composite liquid microecological preparation for feeding the poultry in the table 1;
B. inoculating lactobacillus plantarum LP45 into a fermentation medium a, mixing, performing anaerobic fermentation at 37 ℃ for 16 hours, wherein the inoculation amount is 5% of the mass of the fermentation medium a, and the pH value is controlled at 5.0, so as to obtain lactobacillus plantarum LP45 fermentation liquor;
inoculating bacillus subtilis and bacillus licheniformis to a spore culture medium, inoculating saccharomyces cerevisiae to a fermentation culture medium b, and performing aerobic fermentation at 37 ℃ for 48h to obtain micro-fermentations of the bacillus subtilis, the bacillus licheniformis and the saccharomyces cerevisiae; wherein the inoculation amount of the bacillus subtilis and the bacillus licheniformis is 5% of the mass of the spore culture medium, and the inoculation amount of the saccharomyces cerevisiae is 5% of the mass of the fermentation culture medium b;
C. and D, adding brown granulated sugar into the lactobacillus plantarum LP45 fermentation liquor obtained in the step B, adding micro-fermentation products of bacillus subtilis, bacillus licheniformis and saccharomyces cerevisiae, mixing, performing anaerobic fermentation at 37 ℃ for 85 hours, stirring for 5min every 5 hours, and stopping fermentation when the pressure is higher than 0.1 MPa, the exhaust pressure is reduced to 0.025 MPa, and the pH value is reduced to 4.0.
Comparative example 1
Compared to example 1, the difference is only in step B and step C:
inoculating lactobacillus plantarum LP45, bacillus subtilis, bacillus licheniformis and saccharomyces cerevisiae into a fermentation medium b, mixing, and performing anaerobic fermentation at 37 ℃ for 48 hours, wherein the inoculation amount is 5% of the mass of the fermentation medium b, so as to obtain a fermentation liquid; adding brown granulated sugar into the obtained fermentation liquid, mixing, performing anaerobic fermentation at 37 deg.C for 85 hr, stirring every 5 hr for 5min, discharging gas until the pressure is higher than 0.1 MPa and the pH value is reduced to 0.025 MPa, and stopping fermentation.
Comparative example 2
Compared to example 1, the difference is only in step B and step C:
inoculating lactobacillus plantarum LP45, bacillus subtilis, bacillus licheniformis and saccharomyces cerevisiae into a fermentation medium a, mixing, performing anaerobic fermentation at 37 ℃ for 16 hours, wherein the inoculation amount is 5% of the mass of the fermentation medium a, and the pH value is controlled at 5.0 to obtain a fermentation liquid; adding brown granulated sugar into the obtained fermentation liquid, mixing, performing anaerobic fermentation at 37 deg.C for 85 hr, stirring every 5 hr for 5min, discharging gas until the pressure is higher than 0.1 MPa and the pH value is reduced to 0.025 MPa, and stopping fermentation.
Comparative example 3
Compared to example 1, the difference is only in step B and step C:
inoculating bacillus subtilis and bacillus licheniformis to a spore culture medium, inoculating lactobacillus plantarum LP45 and saccharomyces cerevisiae to a fermentation culture medium b, and carrying out aerobic fermentation at 37 ℃ for 48h to obtain micro-fermentations of the bacillus subtilis, the bacillus licheniformis, the lactobacillus plantarum LP45 and the saccharomyces cerevisiae; wherein the inoculation amounts of the bacillus subtilis and the bacillus licheniformis are both 5 percent of the mass of the spore culture medium, and the inoculation amounts of the lactobacillus plantarum LP45 and the saccharomyces cerevisiae are 5 percent of the mass of the fermentation culture medium b;
adding brown granulated sugar into the obtained micro-fermentation product of Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum LP45, and Saccharomyces cerevisiae, mixing, anaerobic fermenting at 37 deg.C for 85 hr, stirring every 5 hr for 5min, discharging gas to 0.025 MPa when the pressure is higher than 0.1 MPa, and reducing pH to 4.0, and terminating fermentation.
Comparative example 4
Compared to example 1, the difference is only in step B and step C:
inoculating bacillus subtilis and bacillus licheniformis to a spore culture medium, inoculating lactobacillus plantarum LP45 and saccharomyces cerevisiae to a fermentation culture medium a, and carrying out aerobic fermentation at 37 ℃ for 48h to obtain micro-fermentations of the bacillus subtilis, the bacillus licheniformis, the lactobacillus plantarum LP45 and the saccharomyces cerevisiae; wherein the inoculation amounts of the bacillus subtilis and the bacillus licheniformis are both 5 percent of the mass of the spore culture medium, and the inoculation amounts of the lactobacillus plantarum LP45 and the saccharomyces cerevisiae are 5 percent of the mass of the fermentation culture medium a;
adding brown granulated sugar into the obtained micro-fermentation product of Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum LP45, and Saccharomyces cerevisiae, mixing, anaerobic fermenting at 37 deg.C for 85 hr, stirring every 5 hr for 5min, discharging gas to 0.025 MPa when the pressure is higher than 0.1 MPa, and reducing pH to 4.0, and terminating fermentation.
TABLE 1
Second, bacteriostatic effect test
1. Example 1 zone of inhibition experiment of long-acting symbiotic complex liquid microecologics (hereinafter referred to as SD-3301 product), the results are shown in table 2 below.
TABLE 2 diameter of zone of inhibition (unit: cm) of the products of example 1 and comparative examples 1 to 2
As can be seen from the bacteriostatic effect graphs of figures 1-4 and the table 2, the diameters of the bacteriostatic circle of the SD-3301 product are all more than 2cm and significantly more than 0.8cm, which indicates that the product can significantly inhibit the reproduction of intestinal pathogenic bacteria such as escherichia coli, salmonella and the like, is beneficial to maintaining intestinal health and reducing the incidence rate of enteritis and diarrhea, while the comparative example does not adopt the preparation method of the invention, and the bacteriostatic effect of the obtained product is reduced.
Third, acid and bile salt resistance experiment
As can be seen from fig. 5 and 6, the SD-3301 product has very high resistance to artificial gastric juice and artificial intestinal juice, and after 4 hours of acidic condition treatment, the survival rate of more than 90% can still play a role in colonization and probiotic in the intestinal tract; the survival rate of the treated ox bile salt is 80 percent when the treated ox bile salt is 0.3 percent, and enough viable bacteria can reach the intestinal tract for permanent planting to play the probiotic effect. The lowest pH range of the poultry intestinal tract is 3-4, and the acid resistance experiment in figure 2 shows that the product can survive in the poultry intestinal tract and keep the activity to play a role.
Fourth, the experiment of the stability of viable count
As can be seen from FIG. 7, the SD-3301 product of example 1 was left standing at room temperature and under the sun for 90 days, and from 0 to 10 days, the viable count was in a state of increasing continuously, and at the later stage, the viable count gradually decreased with the passage of time, and the tendency of decreasing after approximately decreasing to the initial point was back increased. Under the insolation environment, the viable count keeps relatively stable without great change, which shows that the product of the invention has better stability and can be adapted to different environments for storage.
Fifth, application implementation
Firstly, the long-acting symbiotic composite liquid microecological preparation for poultry feeding in the embodiment 1 is placed in a storage tank, the temperature of the storage tank is slightly higher than the temperature of a use scene, the storage tank is placed for 7-10 days, the pH value and the viable count are periodically detected, and filling is carried out after the stability.
The use method of the long-acting symbiotic composite liquid microecological preparation for poultry feeding comprises the following steps:
1) respectively setting a control group and an experimental group
The broiler chickens of the control group are normally fed without adding any microecologics, and the broiler chickens of the experimental group are normally fed with SD-3301 drinking water.
Experimental group SD-3301 was used in stages: the whole cultivation process is divided into 3 sections, and water is completely drunk after 2-4 hours of concentration each time.
TABLE 3 SD-3301 methods of use
| Age of day | Dosage of |
| 5-8 days old | 0.5L/ten thousand feather/day |
| 13-19 days old | 1L/ten thousand feather/day |
| 32-40 days old | 2L/ten thousand feather/day |
1. Broiler gut health results are shown in table 4.
TABLE 4 influence of SD-3301 on intestinal tracts of broilers
2. As can be seen from the intestinal structure data figure 8, the intestinal villus length/crypt depth of the experimental group is improved by 8.56%, which shows that the SD-3301 product can improve the intestinal structure of poultry and improve the digestion and absorption capacity.
3. Intestinal flora
TABLE 5 influence of probiotic SD-3301 on the number of lactobacilli and coliform bacteria in fecal coliform
As shown in the comparison between the rectal intestinal flora amounts of the experimental group and the control group in the table 5, the SD-3301 probiotics have the effects of maintaining and promoting the reproduction of intestinal probiotics and inhibiting the growth of pathogenic bacteria.
4. Intestinal immunity
TABLE 6 influence of probiotic SD-3301 on broiler antibody levels
As can be seen from the data of the SD-3301 probiotics on the antibody level of the broiler chicken in the table 6, after the microecological product is used, the antibody levels of ND and H9 are obviously improved compared with the control group, which indicates that the SD-3301 probiotics can stimulate the mucosal immunity function and improve the nonspecific immunity, and in addition, the SD-3301 probiotics generate a large amount of acidic substances such as lactic acid and acetic acid in the digestive tract, so that the pH value at the rear end of the digestive tract is effectively reduced, the growth and the propagation of pathogenic bacteria such as escherichia coli and salmonella are inhibited, the vertical transmission of salmonella is prevented, and the occurrence of enteritis is reduced.
TABLE 7 influence of SD-3301 on the level of immunity
As can be seen from Table 7, the antibodies in the experimental group are all higher than those in the control group, the differences are all obvious (p is less than 0.05), the use before and after immunization can obviously improve the immune response, improve the vaccine titer, and accelerate the generation and maintenance of the antibodies, and the probiotic bacteria components act on immune organs in the form of immune adjuvants to promote the immune response of the organism, and the organism is in a high-level immune response state after young birds feed SD-3301 probiotics.
5. Experiment for improving feces
The diarrhea incidence of the experimental group is reduced by 94% compared with the control group, the diarrhea and overfeeding conditions are obviously reduced, the feed utilization rate is improved, and the digestion and absorption are promoted, as shown in figures 9-10.
6. Average weight gain
From FIG. 11, the weight of the experimental group was 5.57 jin, and the weight of the control group was 5.25 jin. The SD-3301 product can improve the weight gain of chicken and improve the production performance.
7. Meat ratio of materials
From FIG. 12, it can be seen that the average feed-meat ratio of the SD-3301 experimental group is reduced by 0.02 compared with the control group; the SD-3301 product is shown to be capable of regulating intestinal tracts of broiler chickens, promoting nutrient absorption and improving the utilization rate of feed.
8. Increase survival rate
As can be seen from FIG. 13, the survival rate of the broiler chickens in the SD-3301 experimental group is 1.96% higher than that of the broiler chickens in the control group, which indicates that the product can significantly improve the physique of the broiler chickens, improve the health conditions of the broiler chickens and reduce the morbidity of the broiler chickens.
9. Weight of chicken
TABLE 8 hairy Chicken weight
| Experiment grouping | Hairy chicken weight (kilogram) |
| Control group | 87363 |
| Experimental group | 90658 |
As shown in Table 8, the total weight of the experimental group was 3295kg higher than that of the control group. The SD-3301 product is shown to be capable of increasing the weight of chickens, the weight of the chickens reflects the growth performance of chicken flocks, and the larger the weight is, the better the growth performance of the chickens is, and the higher the income is.
10. Input-output ratio
TABLE 9 use of the difference in input-output ratio between the SD-3301 experimental group and the control group
The data in the table 9 show that the income of the SD-3301 product is increased by 7.8% with the control group under the same investment cost, which shows that the SD-3301 product can significantly increase the slaughter weight of the broiler chickens, reduce the feed-meat ratio, increase the survival rate of the broiler chickens and bring higher economic benefit for farmers.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions, improvements, etc. within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. The long-acting symbiotic composite liquid microecological preparation for poultry feeding is characterized by comprising, by mass, 5-10 parts of lactobacillus plantarum LP45, 1-5 parts of bacillus subtilis, 1-5 parts of bacillus licheniformis, 2-3 parts of saccharomyces cerevisiae, 11-16 parts of brown granulated sugar and 30-40 parts of water.
2. The long-acting symbiotic composite liquid microecological preparation for poultry feeding according to claim 1, wherein the viable count of the long-acting symbiotic composite liquid microecological preparation is not less than 5 x 108cfu/mL。
3. The preparation method of the long-acting symbiotic composite liquid microecological preparation for poultry feeding is characterized by comprising the following steps of:
A. preparing the components according to the formula of the long-acting symbiotic composite liquid microecological preparation for the poultry feed according to claim 1;
B. inoculating lactobacillus plantarum LP45 into a fermentation medium a, mixing, performing anaerobic fermentation at 37 ℃ for 16 hours, and controlling the pH value to be 5.0 to obtain lactobacillus plantarum LP45 fermentation liquor;
inoculating bacillus subtilis and bacillus licheniformis to a spore culture medium, inoculating saccharomyces cerevisiae to a fermentation culture medium b, and performing aerobic fermentation at 37 ℃ for 48h to obtain micro-fermentations of the bacillus subtilis, the bacillus licheniformis and the saccharomyces cerevisiae;
C. and D, adding brown granulated sugar into the lactobacillus plantarum LP45 fermentation liquor obtained in the step B, adding micro-fermentation products of bacillus subtilis, bacillus licheniformis and saccharomyces cerevisiae, mixing, performing anaerobic fermentation at 37 ℃ for 85 hours, stirring for 5min every 5 hours, reducing the pH value to 4.0, and stopping fermentation.
4. The method for preparing the long-acting symbiotic composite liquid microecological preparation for the poultry feed according to claim 3, wherein the lactobacillus plantarum LP45 inoculation amount in the step B is 5% of the mass of the fermentation medium a.
5. The method for preparing the long-acting symbiotic composite liquid microecological preparation for the poultry feed according to claim 3, wherein the inoculation amount of the bacillus subtilis and the bacillus licheniformis in the step B is 5% of the mass of the spore culture medium, and the inoculation amount of the saccharomyces cerevisiae is 5% of the mass of the fermentation culture medium B.
6. The method for preparing the long-acting symbiotic composite liquid microecological preparation for the poultry feed according to claim 3, wherein the fermentation medium a in the step B comprises, by mass, 1% of beef extract powder, 0.5% of yeast extract, 1% of peptone, 2% of glucose and K2HPO40.2%, sodium acetate 0.5%, MgSO4·7H2O 0.02％，MnSO4·4H20.005% of O, 80.1% of Tween, 0.2% of triammonium citrate and the balance of water; the fermentation medium a is sterilized, and the pH value is 6.2-6.6 before sterilization.
7. The method for preparing the long-acting symbiotic composite liquid microecological preparation for the poultry feed according to claim 3, wherein the fermentation medium B in the step B comprises 1% of yeast extract, 2% of peptone, 2% of glucose and the balance of water by mass percentage; sterilizing the fermentation culture medium b, wherein the pH value is 6.2-6.6 before sterilization;
according to the mass percentage, the spore culture medium in the step B comprises 1% of peptone, 0.3% of beef extract powder, 0.5% of sodium chloride and the balance of water; the spore culture medium is sterilized, and the pH value is 7.0-7.3 before sterilization.
8. The method for preparing the long-acting symbiotic composite liquid microecological preparation for the poultry feed according to claim 3, wherein in the step C, when the pressure is higher than 0.1 MPa, the exhaust pressure is reduced to 0.025 MPa.
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